Patent Publication Number: US-2023133147-A1

Title: ANGIOTENSIN-CONVERTING ENZYME 2 (ACE2) iRNA COMPOSITIONS AND METHODS OF USE THEREOF

Description:
RELATED APPLICATIONS 
     This application is a 35 § U.S.C. 111(a) continuation application which claims the benefit of priority to PCT/US2021/024047, filed on Mar. 25, 2021, which, in turn, claims the benefit of priority to U.S. Provisional Application No. 63/006,383, filed on Apr. 7, 2020; U.S. Provisional Application No. 63/050,135, filed on Jul. 10, 2020; and U.S. Provisional Application No. 63/125,023, filed on Dec. 14, 2020. The entire contents of each of the foregoing applications are incorporated herein by reference. 
    
    
     BACKGROUND OF THE INVENTION 
     Coronaviruses (CoV) are a large family of viruses that cause diseases in mammals and birds. Coronaviruses constitute the subfamily Orthocoronavirinae, in the family Coronaviridae. They are enveloped viruses with a positive-sense single-stranded RNA genome and a nucleocapsid of helical symmetry. The genome size of coronaviruses ranges from approximately 27 to 34 kilobases. The name coronavirus is derived from the Latin corona, meaning “crown” or “halo”, which refers to the characteristic appearance reminiscent of a crown or a solar corona around the virions (virus particles) when viewed under two-dimensional transmission electron microscopy, due to the surface covering in club-shaped protein spikes. 
     The first step of viral infection is viral entry into host cells. The spike (S) protein of coronaviruses facilitates viral entry into target cells. Entry depends on binding of the surface unit, S1, of the S protein to a cellular receptor, which facilitates viral attachment to the surface of target cells. In addition, viral entry into cells requires S protein priming by host cellular proteases, which entails S protein cleavage at the S1/S2 and the S2′ site and allows fusion of viral and cellular membranes, a process driven by the S2 subunit. Previous studies have shown that SARS-CoV-S engages angiotensin-converting enzyme 2 (ACE2) as the entry receptor (Li W, et al.,  Nature  426, 450-454, 2003) and employs the cellular serine protease TMPRSS2 for S protein priming (Glowacka I., et al.,  J. Virol.  85, 4122-4134, 2011; Matsuyama S. et al.,  J. Virol.  84, 12658-12664, 2010; Shulla A., et al.,  J. Virol.  85, 873-882, 2011). The SARS-CoV-S/ACE2 interface has been elucidated at the atomic level, and the efficiency of ACE2 usage was found to be a key determinant of SARS-CoV transmissibility (Li F, et al.,  Science  309, 1864-1868, 2005; Li W. et al.,  EMBO J.  24, 1634-1643, 2005). It has been shown recently that host cell entry of SARS-CoV-2 also depends on the SARS-CoV receptor, ACE2, and that TMPRSS2 is also employed by SARS-CoV-2 for S protein priming (Hoffmann M. at al.,  Cell  181, 1-10, 2020). 
     Coronaviruses can cause illness ranging from the common cold to more severe diseases. For example, infections with the human coronavirus strains CoV-229E, CoV-OC43, CoV-NL63 and CoV-HKU1 usually result in mild, self-limiting upper respiratory tract infections, such as a common cold, e.g., runny nose, sneezing, headache, cough, sore throat or fever (Zumla A. et al.,  Nature Reviews Drug Discovery  15(5): 327-47, 2016; Cheng V. C., et al.,  Clin. Microbial. Rev.  20: 660-694, 2007; Chan J. F. et al.,  Clin. Microbial. Rev.  28: 465-522, 2015). Other infections may result in more severe diseases such as Middle East Respiratory Syndrome (MERS-CoV) and Severe Acute Respiratory Syndrome (SARS-CoV), diseases associated with pneumonia, severe acute respiratory syndrome, kidney failure and death. 
     MERS-CoV and SARS-CoV have received global attention over the past decades owing to their ability to cause community and health-care-associated outbreaks of severe infections in human populations. MERS-CoV is a viral respiratory disease that was first reported in Saudi Arabia in 2012 and has since spread to more than 27 other countries, according to the World Health Organization (de Groot, R. J. et al.,  J. Virol.  87: 7790-7792, 2013). SARS was first reported in Asia in 2003, and quickly spread to about two dozen countries before being contained after about four months (Lee N. et al.,  N. Engl. J. Med.  348: 1986-1994, 2003; Peiris J. S. et al.,  Lancet  36: 1319-1325, 2003). Detailed investigations found that SARS-CoV was transmitted from civet cats to humans and MERS-CoV from dromedary camels to humans (Cheng V. C., et al.,  Clin. Microbial. Rev.  20: 660-694, 2007; Chan J. F. et al.,  Clin. Microbial. Rev.  28: 465-522, 2015). 
     A recent outbreak of respiratory disease caused by a novel coronavirus, severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), was first identified in Wuhan City, China. This disease, named by the World Health Organization as coronavirus disease 2019 (“COVID-19”), presents a major threat to public health worldwide. As of Mar. 26, 2020, there were more than 529,000 confirmed cases and 23,000 deaths across the world. 
     Coronaviruses pose major challenges to clinical management because many questions regarding transmission and control remain unanswered. Moreover, there is currently no vaccine to prevent infections by coronavirus, and there are no specific antiviral treatments available or proven to be effective to treat or prevent coronavirus infection in subjects. Given the critical role that ACE2 plays in the first step of viral infection, ACE2 constitutes a target for antiviral treatment. 
     Accordingly, there exists an immediate need for an agent that can selectively and efficiently silence the ACE2 gene using the cell&#39;s own RNAi machinery that has both high biological activity and in vivo stability, and that can effectively inhibit expression of a target TMPSS2 gene. 
     BRIEF SUMMARY OF THE INVENTION 
     The present disclosure provides RNAi agent compositions which effect the RNA-induced silencing complex (RISC)-mediated cleavage of RNA transcripts of a gene encoding Angiotensin-Converting Enzyme 2 (ACE2). The ACE2 gene may be within a cell, e.g., a cell within a subject, such as a human. The present disclosure also provides methods of using the RNAi agent compositions of the disclosure for inhibiting the expression of an ACE2 gene or for treating a subject who would benefit from inhibiting or reducing the expression of an ACE2 gene, e.g., a subject having an ACE2-associated disorder, e.g., a subject having a coronavirus infection, e.g., a subject having Severe Acute Respiratory Syndrome 2 (SARS-CoV-2; COVID-19), Severe Acute Respiratory Syndrome (SARS-CoV), or Middle East Respiratory Syndrome (MERS-CoV), or a subject at risk of developing a coronavirus infection, e.g., during an epidemic or pandemic. 
     Accordingly, in one aspect, the instant disclosure provides a double stranded ribonucleic acid (dsRNA) agent for inhibiting expression of an angiotensin-converting enzyme 2 (ACE2) gene, wherein the dsRNA agent comprises a sense strand and an antisense strand forming a double stranded region, wherein the sense strand comprises a nucleotide sequence comprising at least 15 contiguous nucleotides, with 0, 1, 2, or 3 mismatches, of a portion of the nucleotide sequence of SEQ ID NO:1, or a nucleotide sequence having at least 90% nucleotide sequence identity to a portion of the nucleotide sequence of SEQ ID NO:1, and the antisense strand comprises a nucleotide sequence comprising at least 15 contiguous nucleotides, with 0, 1, 2, or 3 mismatches, of the corresponding portion of the nucleotide sequence of SEQ ID NO:7, or a nucleotide sequence having at least 90% nucleotide sequence identity to a portion of the nucleotide sequence of SEQ ID NO:7; and wherein the sense strand or the antisense strand is conjugated to one or more lipophilic moieties. 
     In one aspect, the present invention provides a double stranded ribonucleic acid (dsRNA) agent for inhibiting expression of an ACE2 gene in a cell, comprising a sense strand and an antisense strand forming a double stranded region, wherein the antisense strand comprises a region complementary to part of an mRNA encoding an ACE2 gene (SEQ ID NO:1), wherein each strand independently is 14 to 30 nucleotides in length; and wherein the sense strand or the antisense strand is conjugated to one or more lipophilic moieties. 
     In yet another aspect, the present invention provides a double stranded RNAi agent for inhibiting expression of an ACE2 gene in a cell, comprising a sense strand and an antisense strand forming a double stranded region, wherein the antisense strand comprises at least 15 contiguous nucleotides differing by no more than 3 nucleotides from any one of the antisense nucleotide sequences in any one of Tables 2-5, wherein each strand independently is 14 to 30 nucleotides in length; and wherein the sense strand or the antisense strand is conjugated to one or more lipophilic moieties. 
     In one embodiment, the sense strand or the antisense strand is a sense strand or an antisense strand selected from the group consisting of any of the sense strands and antisense strands in any one of Tables 2-5. 
     In one aspect, the present invention provides a double stranded RNAi agent for inhibiting expression of an ACE2 gene in a cell, wherein said dsRNA comprises a sense strand and an antisense strand forming a double stranded region, wherein the sense strand comprises a nucleotide sequence comprising at least 15 contiguous nucleotides, with 0, 1, 2, or 3 mismatches, of nucleotides 1695-1745, 1695-1735, 1695-1732, 1700-1745, 1700-1735, 1704-1732 of SEQ ID NO:1, or a nucleotide sequence having at least 90% nucleotide sequence identity to nucleotides 1695-1745, 1695-1735, 1695-1732, 1700-1745, 1700-1735, 1704-1732, and the antisense strand comprises a nucleotide sequence comprising at least 15 contiguous nucleotides, with 0, 1, 2, or 3 mismatches, of the corresponding portion of the nucleotide sequence of SEQ ID NO:7, or a nucleotide sequence having at least 90% nucleotide sequence identity to a portion of the nucleotide sequence of SEQ ID NO:7; and wherein the sense strand or the antisense strand is conjugated to one or more lipophilic moieties. 
     In one embodiment, the antisense strand comprises at least 15 contiguous nucleotides differing by no more than three nucleotides from any one of the antisense strand nucleotide sequences of a duplex selected from the group consisting of AD-1230825, AD-1230843, and AD-1230934. 
     In one embodiment, the sense strand comprises at least 15 contiguous nucleotides differing by no more than three nucleotides from any one of the sense strand nucleotide sequences of a duplex selected from the group consisting of AD-1230825, AD-1230843, and AD-1230934. 
     In one embodiment, both the sense strand and the antisense strand is conjugated to one or more lipophilic moieties. 
     In one embodiment, the lipophilic moiety is conjugated to one or more positions in the double stranded region of the dsRNA agent. 
     In one embodiment, the lipophilic moiety is conjugated via a linker or a carrier. 
     In one embodiment, lipophilicity of the lipophilic moiety, measured by logKow, exceeds 0. 
     In one embodiment, the hydrophobicity of the double-stranded RNAi agent, measured by the unbound fraction in a plasma protein binding assay of the double-stranded RNAi agent, exceeds 0.2. 
     In one embodiment, the plasma protein binding assay is an electrophoretic mobility shift assay using human serum albumin protein. 
     In one embodiment, the dsRNA agent comprises at least one modified nucleotide. 
     In one embodiment, no more than five of the sense strand nucleotides and no more than five of the nucleotides of the antisense strand are unmodified nucleotides 
     In another embodiment, all of the nucleotides of the sense strand and all of the nucleotides of the antisense strand comprise a modification. 
     In one embodiment, at least one of the modified nucleotides is selected from the group a deoxy-nucleotide, a 3′-terminal deoxy-thymine (dT) nucleotide, a 2′-O-methyl modified nucleotide, a 2′-fluoro modified nucleotide, a 2′-deoxy-modified nucleotide, a locked nucleotide, an unlocked nucleotide, a conformationally restricted nucleotide, a constrained ethyl nucleotide, an abasic nucleotide, a 2′-amino-modified nucleotide, a 2′-O-allyl-modified nucleotide, 2′-C-alkyl-modified nucleotide, a 2′-methoxyethyl modified nucleotide, a 2′-O-alkyl-modified nucleotide, a morpholino nucleotide, a phosphoramidate, a non-natural base comprising nucleotide, a tetrahydropyran modified nucleotide, a 1,5-anhydrohexitol modified nucleotide, a cyclohexenyl modified nucleotide, a nucleotide comprising a 5′-phosphorothioate group, a nucleotide comprising a 5′-methylphosphonate group, a nucleotide comprising a 5′ phosphate or 5′ phosphate mimic, a nucleotide comprising vinyl phosphonate, a nucleotide comprising adenosine-glycol nucleic acid (GNA), a nucleotide comprising thymidine-glycol nucleic acid (GNA) S-Isomer, a nucleotide comprising 2-hydroxymethyl-tetrahydrofurane-5-phosphate, a nucleotide comprising 2′-deoxythymidine-3′phosphate, a nucleotide comprising 2′-deoxyguanosine-3′-phosphate, a 2′-0 hexadecyl nucleotide, a nucleotide comprising a 2′-phosphate, a cytidine-2′-phosphate nucleotide, a guanosine-2′-phosphate nucleotide, a 2′ hexadecyl-cytidine-3′-phosphate nucleotide, a 2′-O-hexadecyl-adenosine-3′-phosphate nucleotide, a 2′-O-hexadecyl-guanosine-3′-phosphate nucleotide, a 2′-O-hexadecyl-uridine-3′-phosphate nucleotide, a a 5′-vinyl phosphonate (VP), a 2′-deoxyadenosine-3′-phosphate nucleotide, a 2′-deoxycytidine-3′-phosphate nucleotide, a 2′-deoxyguanosine-3′-phosphate nucleotide, a 2′-deoxythymidine-3′-phosphate nucleotide, a 2′-deoxyuridine nucleotide, and a terminal nucleotide linked to a cholesteryl derivative and a dodecanoic acid bisdecylamide group; and combinations thereof. 
     In another embodiment, modified nucleotide is selected from the group consisting of a 2′-deoxy-2′-fluoro modified nucleotide, a 2′-deoxy-modified nucleotide, 3′-terminal deoxy-thymine nucleotides (dT), a locked nucleotide, an abasic nucleotide, a 2′-amino-modified nucleotide, a 2′-alkyl-modified nucleotide, a morpholino nucleotide, a phosphoramidate, and a non-natural base comprising nucleotide. 
     In another embodiment, the modified nucleotide comprises a short sequence of 3′-terminal deoxy-thymine nucleotides (dT). 
     In yet another embodiment, the modifications on the nucleotides are 2′-O-methyl modifications, 2′-deoxy-modifications, 2′fluoro modifications, 5′-vinyl phosphonate (VP) modification, and 2′-O hexadecyl nucleotide modifications. 
     In one embodiment, the dsRNA agent further comprises at least one phosphorothioate internucleotide linkage. 
     In one embodiment, the dsRNA agent comprises 6-8 phosphorothioate internucleotide linkages. 
     In one embodiment, each strand is no more than 30 nucleotides in length. 
     In one embodiment, at least one strand comprises a 3′ overhang of at least 1 nucleotide. 
     In another embodiment, at least one strand comprises a 3′ overhang of at least 2 nucleotides. 
     The double stranded region may be 15-30 nucleotide pairs in length; 17-23 nucleotide pairs in length; 17-25 nucleotide pairs in length; 23-27 nucleotide pairs in length; 19-21 nucleotide pairs in length; or 21-23 nucleotide pairs in length. 
     Each strand of the dsRNA agent may be has 19-30 nucleotides in length; 19-23 nucleotides in length; or 21-23 nucleotides in length. 
     In one embodiment, one or more lipophilic moieties are conjugated to one or more internal positions on at least one strand. 
     In one embodiment, the one or more lipophilic moieties are conjugated to one or more internal positions on at least one strand via a linker or carrier. 
     In one embodiment, the internal positions include all positions except the terminal two positions from each end of the at least one strand. 
     In another embodiment, the internal positions include all positions except the terminal three positions from each end of the at least one strand. 
     In another embodiment, the internal positions exclude a cleavage site region of the sense strand. 
     In yet another embodiment, the internal positions include all positions except positions 9-12, counting from the 5′-end of the sense strand. 
     In one embodiment, the internal positions include all positions except positions 11-13, counting from the 3′-end of the sense strand. 
     In one embodiment, the internal positions exclude a cleavage site region of the antisense strand. 
     In one embodiment, the internal positions include all positions except positions 12-14, counting from the 5′-end of the antisense strand. 
     In one embodiment, the internal positions include all positions except positions 11-13 on the sense strand, counting from the 3′-end, and positions 12-14 on the antisense strand, counting from the 5′-end. 
     In one embodiment, the one or more lipophilic moieties are conjugated to one or more of the internal positions selected from the group consisting of positions 4-8 and 13-18 on the sense strand, and positions 6-10 and 15-18 on the antisense strand, counting from the 5′ end of each strand. 
     In one embodiment, the one or more lipophilic moieties are conjugated to one or more of the internal positions selected from the group consisting of positions 5, 6, 7, 15, and 17 on the sense strand, and positions 15 and 17 on the antisense strand, counting from the 5′-end of each strand. 
     In one embodiment, the positions in the double stranded region exclude a cleavage site region of the sense strand. 
     In one embodiment, the sense strand is 21 nucleotides in length, the antisense strand is 23 nucleotides in length, and the lipophilic moiety is conjugated to position 21, position 20, position 15, position 1, position 7, position 6, or position 2 of the sense strand or position 16 of the antisense strand. 
     In one embodiment, the lipophilic moiety is conjugated to position 21, position 20, position 15, position 1, or position 7 of the sense strand. 
     In one embodiment, the lipophilic moiety is conjugated to position 21, position 20, or position 15 of the sense strand. 
     In one embodiment, the lipophilic moiety is conjugated to position 20 or position 15 of the sense strand. 
     In one embodiment, the lipophilic moiety is conjugated to position 16 of the antisense strand. 
     In one embodiment, the lipophilic moiety is an aliphatic, alicyclic, or polyalicyclic compound. 
     In one embodiment, the lipophilic moiety is selected from the group consisting of lipid, cholesterol, retinoic acid, cholic acid, adamantane acetic acid, 1-pyrene butyric acid, dihydrotestosterone, 1,3-bis-O(hexadecyl)glycerol, geranyloxyhexyanol, hexadecylglycerol, borneol, menthol, 1,3-propanediol, heptadecyl group, palmitic acid, myristic acid, O3-(oleoyl)lithocholic acid, O3-(oleoyl)cholenic acid, dimethoxytrityl, or phenoxazine. 
     In one embodiment, the lipophilic moiety contains a saturated or unsaturated C4-C30 hydrocarbon chain, and an optional functional group selected from the group consisting of hydroxyl, amine, carboxylic acid, sulfonate, phosphate, thiol, azide, and alkyne. 
     In one embodiment, the lipophilic moiety contains a saturated or unsaturated C6-C18 hydrocarbon chain. 
     In one embodiment, the lipophilic moiety contains a saturated or unsaturated C16 hydrocarbon chain. 
     In one embodiment, the saturated or unsaturated C16 hydrocarbon chain is conjugated to position 6, counting from the 5′-end of the strand. 
     In one embodiment, the lipophilic moiety is conjugated via a carrier that replaces one or more nucleotide(s) in the internal position(s) or the double stranded region. 
     In one embodiment, the carrier is a cyclic group selected from the group consisting of pyrrolidinyl, pyrazolinyl, pyrazolidinyl, imidazolinyl, imidazolidinyl, piperidinyl, piperazinyl, [1,3]dioxolanyl, oxazolidinyl, isoxazolidinyl, morpholinyl, thiazolidinyl, isothiazolidinyl, quinoxalinyl, pyridazinonyl, tetrahydrofuranyl, and decalinyl; or is an acyclic moiety based on a serinol backbone or a diethanolamine backbone. 
     In one embodiment, the lipophilic moiety is conjugated to the double-stranded iRNA agent via a linker containing an ether, thioether, urea, carbonate, amine, amide, maleimide-thioether, disulfide, phosphodiester, sulfonamide linkage, a product of a click reaction, or carbamate. 
     In one embodiment, the lipophilic moiety is conjugated to a nucleobase, sugar moiety, or internucleosidic linkage. 
     In one embodiment, the lipophilic moiety or a targeting ligand is conjugated via a bio-cleavable linker selected from the group consisting of DNA, RNA, disulfide, amide, functionalized monosaccharides or oligosaccharides of galactosamine, glucosamine, glucose, galactose, mannose, and combinations thereof. 
     In one embodiment, the 3′ end of the sense strand is protected via an end cap which is a cyclic group having an amine, said cyclic group being selected from the group consisting of pyrrolidinyl, pyrazolinyl, pyrazolidinyl, imidazolinyl, imidazolidinyl, piperidinyl, piperazinyl, [1,3]dioxolanyl, oxazolidinyl, isoxazolidinyl, morpholinyl, thiazolidinyl, isothiazolidinyl, quinoxalinyl, pyridazinonyl, tetrahydrofuranyl, and decalinyl. 
     In one embodiment, the dsRNA agent further comprises a targeting ligand that targets a liver tissue. 
     In one embodiment, the targeting ligand is a GalNAc conjugate. 
     In one embodiment, the dsRNA agent further comprises a terminal, chiral modification occurring at the first internucleotide linkage at the 3′ end of the antisense strand, having the linkage phosphorus atom in Sp configuration, a terminal, chiral modification occurring at the first internucleotide linkage at the 5′ end of the antisense strand, having the linkage phosphorus atom in Rp configuration, and a terminal, chiral modification occurring at the first internucleotide linkage at the 5′ end of the sense strand, having the linkage phosphorus atom in either Rp configuration or Sp configuration. 
     In one embodiment, the dsRNA agent further comprises a terminal, chiral modification occurring at the first and second internucleotide linkages at the 3′ end of the antisense strand, having the linkage phosphorus atom in Sp configuration, a terminal, chiral modification occurring at the first internucleotide linkage at the 5′ end of the antisense strand, having the linkage phosphorus atom in Rp configuration, and a terminal, chiral modification occurring at the first internucleotide linkage at the 5′ end of the sense strand, having the linkage phosphorus atom in either Rp or Sp configuration. 
     In one embodiment, the dsRNA agent further comprises a terminal, chiral modification occurring at the first, second and third internucleotide linkages at the 3′ end of the antisense strand, having the linkage phosphorus atom in Sp configuration, a terminal, chiral modification occurring at the first internucleotide linkage at the 5′ end of the antisense strand, having the linkage phosphorus atom in Rp configuration, and a terminal, chiral modification occurring at the first internucleotide linkage at the 5′ end of the sense strand, having the linkage phosphorus atom in either Rp or Sp configuration. 
     In one embodiment, the dsRNA agent further comprises a terminal, chiral modification occurring at the first, and second internucleotide linkages at the 3′ end of the antisense strand, having the linkage phosphorus atom in Sp configuration, a terminal, chiral modification occurring at the third internucleotide linkages at the 3′ end of the antisense strand, having the linkage phosphorus atom in Rp configuration, a terminal, chiral modification occurring at the first internucleotide linkage at the 5′ end of the antisense strand, having the linkage phosphorus atom in Rp configuration, and a terminal, chiral modification occurring at the first internucleotide linkage at the 5′ end of the sense strand, having the linkage phosphorus atom in either Rp or Sp configuration. 
     In one embodiment, the dsRNA agent further comprises a terminal, chiral modification occurring at the first, and second internucleotide linkages at the 3′ end of the antisense strand, having the linkage phosphorus atom in Sp configuration, a terminal, chiral modification occurring at the first, and second internucleotide linkages at the 5′ end of the antisense strand, having the linkage phosphorus atom in Rp configuration, and a terminal, chiral modification occurring at the first internucleotide linkage at the 5′ end of the sense strand, having the linkage phosphorus atom in either Rp or Sp configuration. 
     In one embodiment, the dsRNA agent further comprises a phosphate or phosphate mimic at the 5′-end of the antisense strand. 
     In one embodiment, the phosphate mimic is a 5′-vinyl phosphonate (VP). 
     In one embodiment, the base pair at the 1 position of the 5′-end of the antisense strand of the duplex is an AU base pair. 
     In one embodiment, the sense strand has a total of 21 nucleotides and the antisense strand has a total of 23 nucleotides. 
     The present invention further provides cells, pharmaceutical compositions for inhibiting expression of an ACE2 gene, and pharmaceutical composition comprising a lipid formulation. comprising the dsRNA agent of the invention. 
     In one aspect, the present invention provides a method of inhibiting expression of an ACE2 gene in a cell. The method includes contacting the cell with the dsRNA agent of the invention, or the pharmaceutical composition of the invention; and maintaining the cell produced in step (a) for a time sufficient to obtain degradation of the mRNA transcript of an ACE2 gene, thereby inhibiting expression of the ACE2 gene in the cell. 
     In one embodiment, the cell is within a subject. 
     In one embodiment, the subject is a human 
     In one embodiment, the expression of the ACE2 gene is inhibited by at least 50%. 
     In one aspect, the present invention provides a method of inhibiting entry of a coronavirus into a cell. The method includes contacting the cell with the dsRNA agent of the invention, or the pharmaceutical composition of the invention; and maintaining the cell produced in step (a) for a time sufficient to obtain degradation of the mRNA transcript of the ACE2 gene, thereby inhibiting entry of the coronavirus into the cell. 
     In one embodiment, the cell is within a subject. 
     In one embodiment, the subject is a human 
     In one embodiment, the expression of the ACE2 gene is inhibited by at least 50%. 
     In one aspect, the present invention provides a method of inhibiting replication of a coronavirus in a cell. The method includes contacting the cell with the dsRNA agent of the invention, or the pharmaceutical composition of the invention; and maintaining the cell produced in step (a) for a time sufficient to obtain degradation of the mRNA transcript of the ACE2 gene, thereby inhibiting replication of the coronavirus in the cell. 
     In one embodiment, the cell is within a subject. 
     In one embodiment, the subject is a human 
     In one embodiment, the expression of the ACE2 gene is inhibited by at least 50%. 
     In another aspect, the present invention provides a method of inhibiting priming of a coronavirus S protein in a cell. The method includes contacting the cell with the dsRNA agent of the invention, or the pharmaceutical composition of the invention; and maintaining the cell produced in step (a) for a time sufficient to obtain degradation of an mRNA transcript of an ACE2 gene, thereby inhibiting priming of a coronavirus S protein in the cell. 
     In one embodiment, the cell is within a subject. 
     In one embodiment, the subject is a human 
     In one embodiment, the expression of the ACE2 gene is inhibited by at least 50%. 
     In one aspect, the present invention provides a method of treating a TNPRSS2-associate disorder, e.g., a subject having a coronavirus infection or at risk of developing or at risk of having a coronavirus infection. The method includes administering to the subject a therapeutically effective amount of the dsRNA agent of the invention, or the pharmaceutical composition of the invention, thereby treating the subject. 
     In one embodiment, the subject is a human 
     In one embodiment, the subject having the coronavirus infection is infected with a severe acute respiratory syndrome (SARS) virus, a Middle East respiratory syndrome (MERS) virus, or a severe acute respiratory syndrome 2 (SARS-2) virus. 
     In one embodiment, the subject at risk of developing a coronavirus infection, e.g., an infection caused by severe acute respiratory syndrome (SARS) virus, a Middle East respiratory syndrome (MERS) virus, or a severe acute respiratory syndrome 2 (SARS-2) virus, is a subject in an epidemic or pandemic. 
     In one embodiment, treating comprises amelioration of at least on sign or symptom of the disease. 
     In one embodiment, the dsRNA agent is administered to the subject at a dose of about 0.01 mg/kg to about 50 mg/kg. 
     In one embodiment, the administration of the dsRNA is pulmonary system administration. 
     In one embodiment, the pulmonary system administration is via oral inhalation or intranasally. 
     In one embodiment, the method reduces the expression of an ACE2 gene in a pulmonary system tissue, e.g., a nasopharynx tissue, an oropharynx tissue, a laryngopharynx tissue, a larynx tissue, a trachea tissue, a carina tissue, a bronchi tissue, a bronchiole tissue, or an alveoli tissue. 
     In one embodiment, the dsRNA agent is administered to the subject subcutaneously. 
     In one embodiment, the method further comprises administering to the subject an additional agent or a therapy suitable for treatment or prevention of a coronavirus-associated disorder. 
     In one embodiment, the additional therapeutic agent is selected from the group consisting of an antiviral agent, an immune stimulator, a therapeutic vaccine, a viral entry inhibitor, and a combination of any of the foregoing. 
     The present invention is further illustrated by the following detailed description. 
    
    
     
       BRIEF DESCRIPTION OF THE DRAWINGS 
         FIG.  1    is a graph depicting the effect of intranasal administration of a AD-1230934 on the body weight of hamsters challenged with SARS-CoV-2. 
     
    
    
     DETAILED DESCRIPTION OF THE INVENTION 
     The present invention provides iRNA compositions which effect the RNA-induced silencing complex (RISC)-mediated cleavage of RNA transcripts of an ACE2 gene. The ACE2 gene may be within a cell, e.g., a cell within a subject, such as a human. The use of these iRNAs enables the targeted degradation of mRNAs of the corresponding gene (an ACE2 gene) in mammals. The present disclosure also provides methods of using the RNAi compositions of the disclosure for inhibiting the expression of an ACE2 gene for treating a subject having a disorder that would benefit from inhibiting or reducing the expression of an ACE2 gene, e.g., an ACE2-associated disorder, e.g., a coronavirus-associated disorder, e.g., a subject having a coronavirus infection, e.g., a subject having Severe Acute Respiratory Syndrome 2 (SARS-CoV-2; COVID-19), Severe Acute Respiratory Syndrome (SARS-CoV), or Middle East Respiratory Syndrome (MERS-CoV), or a subject at risk of a coronavirus infection, e.g., infection by Severe Acute Respiratory Syndrome 2 (SARS-CoV-2; COVID-19), Severe Acute Respiratory Syndrome (SARS-CoV), or Middle East Respiratory Syndrome (MERS-CoV), e.g., during an epidemic or pandemic 
     The iRNAs of the invention include an RNA strand (the antisense strand) having a region which is up to about 30 nucleotides or less in length, e.g., 19-30, 19-29, 19-28, 19-27, 19-26, 19-25, 19-24, 19-23, 19-22, 19-21, 19-20, 20-30, 20-29, 20-28, 20-27, 20-26, 20-25, 20-24,20-23, 20-22, 20-21, 21-30, 21-29, 21-28, 21-27, 21-26, 21-25, 21-24, 21-23, or 21-22 nucleotides in length, which region is substantially complementary to at least part of an mRNA transcript of an ACE2 gene. In certain embodiments, the RNAi agents of the disclosure include an RNA strand (the antisense strand) having a region which is about 21-23 nucleotides in length, which region is substantially complementary to at least part of an mRNA transcript of an ACE2 gene. 
     In certain embodiments, one or both of the strands of the double stranded RNAi agents of the invention is up to 66 nucleotides in length, e.g., 36-66, 26-36, 25-36, 31-60, 22-43, 27-53 nucleotides in length, with a region of at least 19 contiguous nucleotides that is substantially complementary to at least a part of an mRNA of an ACE2 gene. In some embodiments, such iRNA agents having longer length antisense strands preferably may include a second RNA strand (the sense strand) of 20-60 nucleotides in length wherein the sense and antisense strands form a duplex of 18-30 contiguous nucleotides. 
     The use of iRNAs of the invention enables the targeted degradation of the ACE2 mRNAs in mammals. Thus, methods and compositions including these iRNAs are useful for treating a subject having an ACE2-associated disorder, e.g., a coronavirus-associated disorder, e.g., a subject having a coronavirus infection, e.g., a subject having Severe Acute Respiratory Syndrome 2 (SARS-CoV-2; COVID-19), Severe Acute Respiratory Syndrome (SARS-CoV), or Middle East Respiratory Syndrome (MERS-CoV) or treating a subject at risk of an ACE2-associate disorder, e.g., a subject at risk of a coronavirus infection, e.g., infections resulting in Severe Acute Respiratory Syndrome 2 (SARS-CoV-2; COVID-19), Severe Acute Respiratory Syndrome (SARS-CoV), or Middle East Respiratory Syndrome (MERS-CoV), e.g., during an epidemic or pandemic. 
     In certain embodiments, the administration of the dsRNA to a subject results in an improvement in viral load, of lung function, or a stoppage or reduction of the rate of loss of lung function, reduction of fever, reduction of cough. 
     The following detailed description discloses how to make and use compositions containing iRNAs to inhibit the expression of an ACE2 gene s as well as compositions, uses, and methods for treating subjects that would benefit from inhibition and/or reduction of the expression of an ACE2 gene, e.g., subjects susceptible to or diagnosed with an ACE2-associated disorder. 
     I. DEFINITIONS 
     In order that the present invention may be more readily understood, certain terms are first defined. In addition, it should be noted that whenever a value or range of values of a parameter are recited, it is intended that values and ranges intermediate to the recited values are also intended to be part of this invention. 
     The articles “a” and “an” are used herein to refer to one or to more than one (i.e., to at least one) of the grammatical object of the article. By way of example, “an element” means one element or more than one element, e.g., a plurality of elements. 
     The term “including” is used herein to mean, and is used interchangeably with, the phrase “including but not limited to”. 
     The term “or” is used herein to mean, and is used interchangeably with, the term “and/or,” unless context clearly indicates otherwise. 
     The term “about” is used herein to mean within the typical ranges of tolerances in the art. For example, “about” can be understood as about 2 standard deviations from the mean. In certain embodiments, about means±10%. In certain embodiments, about means±5%. When about is present before a series of numbers or a range, it is understood that “about” can modify each of the numbers in the series or range. 
     The term “at least” prior to a number or series of numbers is understood to include the number adjacent to the term “at least”, and all subsequent numbers or integers that could logically be included, as clear from context. For example, the number of nucleotides in a nucleic acid molecule must be an integer. For example, “at least 19 nucleotides of a 21 nucleotide nucleic acid molecule” means that 19, 20, or 21 nucleotides have the indicated property. When at least is present before a series of numbers or a range, it is understood that “at least” can modify each of the numbers in the series or range. 
     As used herein, “no more than” or “less than” is understood as the value adjacent to the phrase and logical lower values or integers, as logical from context, to zero. For example, a duplex with an overhang of “no more than 2 nucleotides” has a 2, 1, or 0 nucleotide overhang. When “no more than” is present before a series of numbers or a range, it is understood that “no more than” can modify each of the numbers in the series or range. As used herein, ranges include both the upper and lower limit. 
     As used herein, methods of detection can include determination that the amount of analyte present is below the level of detection of the method. 
     In the event of a conflict between an indicated target site and the nucleotide sequence for a sense or antisense strand, the indicated sequence takes precedence. 
     In the event of a conflict between a sequence and its indicated site on a transcript or other sequence, the nucleotide sequence recited in the specification takes precedence. 
     As used herein, the term “coronavirus,” (“CoV”; subfamily Coronaviridae, family Coronaviridae, order Nidovirales), refers to a group of highly diverse, enveloped, positive-sense, single-stranded RNA viruses that cause respiratory, enteric, hepatic and neurological diseases of varying severity in a broad range of animal species, including humans. Coronaviruses are subdivided into four genera: Alphacoronavirus, Betacoronavirus (13CoV), Gammacoronavirus and Deltacoronavirus. 
     Any coronavirus that infects humans and animals is encompassed by the term “coronavirus” as used herein. Exemplary coronaviruses encompassed by the term include the coronaviruses that cause a common cold-like respiratory illness, e.g., human coronavirus 229E (HCoV-229E), human coronavirus NL63 (HCoV-NL63), human coronavirus OC43 (HCoV-OC43), and human coronavirus HKU1 (HCoV-HKU1); the coronavirus that causes avian infectious bronchitis virus (IBV); the coronavirus that causes murine hepatitis virus (MHV); the coronavirus that causes porcine transmissible gastroenteritis virus PRCoV; the coronavirus that causes porcine respiratory coronavirus and bovine coronavirus; the coronavirus that causes Severe Acute Respiratory Syndrome (SARS), the coronavirus that causes the Middle East respiratory syndrome (MERS), and the coronavirus that causes Severe Acute Respiratory Syndrome 2 (SARS-CoV-2; COVID-19). 
     The coronavirus (CoV) genome is a single-stranded, non-segmented RNA genome, which is approximately 26-32 kb. It contains 5′-methylated caps and 3′-polyadenylated tails and is arranged in the order of 5′, replicase genes, genes encoding structural proteins (spike glycoprotein (S), envelope protein (E), membrane protein (M) and nucleocapsid protein (N)), polyadenylated tail and then the 3′ end. The partially overlapping 5′-terminal open reading frame 1a/b (ORF1a/b) is within the 5′ two-thirds of the CoV genome and encodes the large replicase polyprotein 1a (pp1a) and pp1ab. These polyproteins are cleaved by papain-like cysteine protease (PLpro) and 3C-like serine protease (3CLpro) to produce non-structural proteins, including RNA-dependent RNA polymerase (RdRp) and helicase (Hel), which are important enzymes involved in the transcription and replication of CoVs. The 3′ one-third of the CoV genome encodes the structural proteins (S, E, M and N), which are essential for virus-cell-receptor binding and virion assembly, and other non-structural proteins and accessory proteins that may have immunomodulatory effects. (Peiris J S., et al., 2003 , Nat. Med.  10 (Suppl. 12): 88-97). 
     As a coronavirus is a positive-sense, single-stranded RNA virus having a 5′ methylated cap and a 3′ polyadenylated tail, once the virus enters the cell and is uncoated, the viral RNA genome attaches to the host cell&#39;s ribosome for direct translation. The host ribosome translates the initial overlapping open reading frame of the virus genome and forms a long polyprotein. The polyprotein has its own proteases which cleave the polyprotein into multiple nonstructural proteins. 
     A number of the nonstructural proteins coalesce to form a multi-protein replicase-transcriptase complex (RTC). The main replicase-transcriptase protein is the RNA-dependent RNA polymerase (RdRp). It is directly involved in the replication and transcription of RNA from an RNA strand. The other nonstructural proteins in the complex assist in the replication and transcription process. The exoribonuclease non-structural protein for instance provides extra fidelity to replication by providing a proofreading function which the RNA-dependent RNA polymerase lacks. 
     One of the main functions of the complex is to replicate the viral genome. RdRp directly mediates the synthesis of negative-sense genomic RNA from the positive-sense genomic RNA. This is followed by the replication of positive-sense genomic RNA from the negative-sense genomic RNA. The other important function of the complex is to transcribe the viral genome. RdRp directly mediates the synthesis of negative-sense subgenomic RNA molecules from the positive-sense genomic RNA. This is followed by the transcription of these negative-sense subgenomic RNA molecules to their corresponding positive-sense mRNAs 
     The replicated positive-sense genomic RNA becomes the genome of the progeny viruses. 
     As use herein, the term “severe acute respiratory syndrome coronavirus” or “SARS-CoV”, refers to a coronavirus that was first discovered in 2003, which causes severe acute respiratory syndrome (SARS). SARS-CoV represents the prototype of a new lineage of coronaviruses capable of causing outbreaks of clinically significant and frequently fatal human disease. The complete genome of SARS-CoV has been identified, as well as common variants thereof. The genome of SARS-CoV is a 29,727-nucleotide polyadenylated RNA, has 11 open reading frames, and 41% of the residues are G or C. The genomic organization is typical of coronaviruses, with the characteristic gene order (5′-replicase (rep), spike (S), envelope (E), membrane (M), nucleocapsid (N)-3′ and short untranslated regions at both termini. The SARS-CoV rep gene, which comprises about two-thirds of the genome, is predicted to encode two polyproteins that undergo co-translational proteolytic processing. There are four open reading frames (ORFs) downstream of rep that are predicted to encode the structural proteins, S, E, M and N. The hemagglutinin-esterase gene, which is present between ORF1b and S in group 2 and some group 3 coronaviruses was not found. 
     The amino acid and complete coding sequences of the SARS-CoV genomes are known may be found in for example, GenBank Accession Nos. AY502923.1; AP006559.1; AP006558.1; AY313906.1; AY345986.1; AY502931.1; AY282752.2; AY559097.1; AY559081.1; DQ182595.1; AY291451.1; AY568539.1; AY613947.1; and AY390556.1, the entire contents of each of which are incorporated herein by reference. 
     The term “SARS-CoV,” as used herein, also refers to naturally occurring RNA sequence variations of the SARS-CoV genome. 
     As use herein, the term “the Middle East respiratory syndrome coronavirus” or “MERS-CoV”, refers to a coronavirus that causes the Middle East respiratory syndrome (MERS), which was first identified in 2012. MERS-CoV is closely related to severe acute respiratory syndrome (SARS) coronavirus (SARS-CoV). Clinically similar to SARS, MERS-CoV infection leads to severe respiratory illness with renal failure. 
     The amino acid and complete coding sequences of the MERS-CoV genomes are known and may be found in for example, GenBank Accession Nos. MK462243.1; MK462244.1; MK462245.1; MK462246.1; MK462247.1; MK462248.1; MK462249.1; MK462250.1; MK462251.1; MK462252.1; MK462253.1; MK462254.1; MK462255.1; MK462256.1; MK483839.1; and MH822886.1, the entire contents of each of which are incorporated herein by reference. 
     The term “MERS-CoV,” as used herein, also refers to naturally occurring RNA sequence variations of the MERS-CoV genome. 
     As use herein, the terms “severe acute respiratory syndrome coronavirus 2,” “SARS-CoV-2,” “2019-nCoV,” refer to the novel coronavirus that caused a pneumonia outbreak first reported in Wuhan, China in December 2019 (“COVID-19”). Phylogenetic analysis of the complete viral genome (29,903 nucleotides) revealed that SARS-CoV-2 was most closely related (89.1% nucleotide similarity similarity) to SARS-CoV. 
     The amino acid and complete coding sequences of the SARS-CoV-2 genomes are known and may be found in for example, the GISAID EpiCoV™ Database (db.cngb.org/gisaid/), including Accession nos. EPI_ISL_402119; EPI_ISL_402120; EPI_ISL_402121; EPI_ISL_402123; EPI_ISL_402124; EPI_ISL_402125; EPI_ISL_402127; EPI_ISL_402128; EPI_ISL_402129; EPI_ISL_402130; EPI_ISL_402132; EPI_ISL_403928; EPI_ISL_403929; EPI_ISL_403930; EPI_ISL_403931; EPI_ISL_403932; EPI_ISL_403933; EPI_ISL_403934; EPI_ISL_403935; EPI_ISL_403936; EPI_ISL_403937; EPI_ISL_403962; EPI_ISL_404228; EPI_ISL_404253; and EPI_ISL_404895, the entire contents of which of which are incorporated herein by reference. 
     The term “SARS-CoV-2,” as used herein, also refers to naturally occurring RNA sequence variations of the SARS-CoV-2 genome. 
     Additional examples of coronavirus genomes and mRNA sequences are readily available using publicly available databases, e.g., GenBank, UniProt, and OMIM. 
     As used herein, the term “Angiotensin I Converting Enzyme 2,” used interchangeably with the term “ACE2,” refers to the well-known gene and polypeptide, also known in the art as Angiotensin-Converting Enzyme Homolog, Angiotensin-Converting Enzyme 2, ACE-Related Carboxypeptidase, Metalloprotease MPROT15, Peptidyl-Dipeptidase A and ACEH. The term “ACE2” includes human ACE2, the amino acid and nucleotide sequences of which may be found in, for example, GenBank Accession No. NM_021804.3 (GI: 1700998533; SEQ ID NO:1) and GenBank Accession No. NM_001371415.1 (GI: 1700998531; SEQ ID NO: 2); mouse ACE2, the amino acid and nucleotide sequence of which may be found in, for example, GenBank Accession No. NM_001130513.1 (GI: 194473666, SEQ ID NO: 3); and GenBank Accession No. NM_027286.4 (GI: 194473665; SEQ ID NO: 4); and rat ACE2, the amino acid and nucleotide sequence of which may be found in, for example, GenBank Accession No. NM_001012006.1 (GI: 58865587; SEQ ID NO: 5). 
     The term “ACE2” also includes  Macaca fascicularis  ACE2, the amino acid and nucleotide sequence of which may be found in, for example, GenBank Accession No. XM_005593037.2 (GI: 982321771; SEQ ID NO: 6). 
     Additional examples of ACE2 mRNA sequences are readily available using, e.g., GenBank, UniProt, OMIM, and the Macaca genome project web site. 
     Exemplary ACE2 nucleotide sequences may also be found in SEQ ID NOs: 1-12. SEQ ID NOs: 7-12 are the reverse complement sequences of SEQ ID NOs: 1-6, respectively. 
     Further information on ACE2 is provided, for example in the NCBI Gene database at www.ncbi.nlm.nih.gov/gene/59272. 
     The entire contents of each of the foregoing GenBank Accession numbers and the Gene database numbers are incorporated herein by reference as of the date of filing this application. 
     The terms “angiotensin converting enzyme 2” and “ACE2,” as used herein, also refer to naturally occurring DNA sequence variations of the ACE2 gene. Numerous sequence variations within the ACE2 gene have been identified and may be found at, for example, NCBI dbSNP and UniProt (see, e.g., www.ncbi.nlm.nih.gov/snp?LinkName=gene_snp&amp;from_uid=5927), the entire contents of which are incorporated herein by reference as of the date of filing this application. 
     As used herein, “target sequence” refers to a contiguous portion of the nucleotide sequence of an mRNA molecule formed during the transcription of an ACE2 gene, including mRNA that is a product of RNA processing of a primary transcription product. In one embodiment, the target portion of the sequence will be at least long enough to serve as a substrate for RNAi-directed cleavage at or near that portion of the nucleotide sequence of an mRNA molecule formed during the transcription of an ACE2 gene. In one embodiment, the target sequence is within the protein coding region of the ACE2 gene. In another embodiment, the target sequence is within the 3′ UTR of the ACE2 gene. 
     The target sequence may be from about 19-36 nucleotides in length, e.g., preferably about 19-30 nucleotides in length. For example, the target sequence can be about 19-30 nucleotides, 19-30, 19-29, 19-28, 19-27, 19-26, 19-25, 19-24, 19-23, 19-22, 19-21, 19-20, 20-30, 20-29, 20-28, 20-27, 20-26, 20-25, 20-24, 20-23, 20-22, 20-21, 21-30, 21-29, 21-28, 21-27, 21-26, 21-25, 21-24, 21-23, or 21-22 nucleotides in length. In some embodiments, the target sequence is about 19 to about 30 nucleotides in length. In other embodiments, the target sequence is about 19 to about 25 nucleotides in length. In still other embodiments, the target sequence is about 19 to about 23 nucleotides in length. In some embodiments, the target sequence is about 21 to about 23 nucleotides in length. Ranges and lengths intermediate to the above recited ranges and lengths are also contemplated to be part of the invention. 
     As used herein, the term “strand comprising a sequence” refers to an oligonucleotide comprising a chain of nucleotides that is described by the sequence referred to using the standard nucleotide nomenclature. 
     “G,” “C,” “A,” “T,” and “U” each generally stand for a nucleotide that contains guanine, cytosine, adenine, thymidine, and uracil as a base, respectively. However, it will be understood that the term “ribonucleotide” or “nucleotide” can also refer to a modified nucleotide, as further detailed below, or a surrogate replacement moiety (see, e.g., Table 1). The skilled person is well aware that guanine, cytosine, adenine, and uracil can be replaced by other moieties without substantially altering the base pairing properties of an oligonucleotide comprising a nucleotide bearing such replacement moiety. For example, without limitation, a nucleotide comprising inosine as its base can base pair with nucleotides containing adenine, cytosine, or uracil. Hence, nucleotides containing uracil, guanine, or adenine can be replaced in the nucleotide sequences of dsRNA featured in the invention by a nucleotide containing, for example, inosine. In another example, adenine and cytosine anywhere in the oligonucleotide can be replaced with guanine and uracil, respectively to form G-U Wobble base pairing with the target mRNA. Sequences containing such replacement moieties are suitable for the compositions and methods featured in the invention. 
     The terms “iRNA”, “RNAi agent,” “iRNA agent,” “RNA interference agent” as used interchangeably herein, refer to an agent that contains RNA as that term is defined herein, and which mediates the targeted cleavage of an RNA transcript via an RNA-induced silencing complex (RISC) pathway. RNA interference (RNAi) is a process that directs the sequence-specific degradation of mRNA. RNAi modulates, e.g., inhibits, the expression of an ACE2 gene in a cell, e.g., a cell within a subject, such as a mammalian subject. 
     In one embodiment, an RNAi agent of the disclosure includes a single stranded RNAi that interacts with a target RNA sequence, e.g., an ACE2 mRNA sequence, to direct the cleavage of the target RNA. Without wishing to be bound by theory it is believed that long double stranded RNA introduced into cells is broken down into double-stranded short interfering RNAs (siRNAs) comprising a sense strand and an antisense strand by a Type III endonuclease known as Dicer (Sharp et al. (2001)  Genes Dev.  15:485). Dicer, a ribonuclease-III-like enzyme, processes these dsRNA into 19-23 base pair short interfering RNAs with characteristic two base 3′ overhangs (Bernstein, et al., (2001)  Nature  409:363). These siRNAs are then incorporated into an RNA-induced silencing complex (RISC) where one or more helicases unwind the siRNA duplex, enabling the complementary antisense strand to guide target recognition (Nykanen, et al., (2001)  Cell  107:309). Upon binding to the appropriate target mRNA, one or more endonucleases within the RISC cleave the target to induce silencing (Elbashir, et al., (2001)  Genes Dev.  15:188). Thus, in one aspect the disclosure relates to a single stranded RNA (ssRNA) (the antisense strand of a siRNA duplex) generated within a cell and which promotes the formation of a RISC complex to effect silencing of the target gene. Accordingly, the term “siRNA” is also used herein to refer to an RNAi as described above. 
     In another embodiment, the RNAi agent may be a single-stranded RNA that is introduced into a cell or organism to inhibit a target mRNA. Single-stranded RNAi agents bind to the RISC endonuclease, Argonaute 2, which then cleaves the target mRNA. The single-stranded siRNAs are generally 15-30 nucleotides and are chemically modified. The design and testing of single-stranded RNAs are described in U.S. Pat. No. 8,101,348 and in Lima et al., (2012)  Cell  150:883-894, the entire contents of each of which are hereby incorporated herein by reference. Any of the antisense nucleotide sequences described herein may be used as a single-stranded siRNA as described herein or as chemically modified by the methods described in Lima et al., (2012)  Cell  150:883-894. 
     In another embodiment, a “RNAi agent” for use in the compositions and methods of the disclosure is a double stranded RNA and is referred to herein as a “double stranded RNAi agent,” “double stranded RNA (dsRNA) molecule,” “dsRNA agent,” or “dsRNA”. The term “dsRNA” refers to a complex of ribonucleic acid molecules, having a duplex structure comprising two anti-parallel and substantially complementary nucleic acid strands, referred to as having “sense” and “antisense” orientations with respect to a target RNA, i.e., an ACE2 mRNA sequence. In some embodiments of the disclosure, a double stranded RNA (dsRNA) triggers the degradation of a target RNA, e.g., an mRNA, through a post-transcriptional gene-silencing mechanism referred to herein as RNA interference or RNAi. 
     In general, a dsRNA molecule can include ribonucleotides, but as described in detail herein, each or both strands can also include one or more non-ribonucleotides, e.g., a deoxyribonucleotide, a modified nucleotide. In addition, as used in this specification, an “RNAi agent” may include ribonucleotides with chemical modifications; an RNAi agent may include substantial modifications at multiple nucleotides. 
     As used herein, the term “modified nucleotide” refers to a nucleotide having, independently, a modified sugar moiety, a modified internucleotide linkage, or a modified nucleobase. Thus, the term modified nucleotide encompasses substitutions, additions or removal of, e.g., a functional group or atom, to internucleoside linkages, sugar moieties, or nucleobases. The modifications suitable for use in the agents of the disclosure include all types of modifications disclosed herein or known in the art. Any such modifications, as used in a siRNA type molecule, are encompassed by “RNAi agent” for the purposes of this specification and claims. 
     In certain embodiments of the instant disclosure, inclusion of a deoxy-nucleotide—which is acknowledged as a naturally occurring form of nucleotide—if present within a RNAi agent can be considered to constitute a modified nucleotide. 
     The duplex region may be of any length that permits specific degradation of a desired target RNA through a RISC pathway, and may range from about 9 to 36 base pairs in length, e.g., about 15-30 base pairs in length, for example, about 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, or 36 base pairs in length, such as about 15-30, 15-29, 15-28, 15-27, 15-26, 15-25, 15-24, 15-23, 15-22, 15-21, 15-20, 15-19, 15-18, 15-17, 18-30, 18-29, 18-28, 18-27, 18-26, 18-25, 18-24, 18-23, 18-22, 18-21, 18-20, 19-30, 19-29, 19-28, 19-27, 19-26, 19-25, 19-24, 19-23, 19-22, 19-21, 19-20, 20-30, 20-29, 20-28, 20-27, 20-26, 20-25, 20-24,20-23, 20-22, 20-21, 21-30, 21-29, 21-28, 21-27, 21-26, 21-25, 21-24, 21-23, or 21-22 base pairs in length. Ranges and lengths intermediate to the above recited ranges and lengths are also contemplated to be part of the invention. 
     The two strands forming the duplex structure may be different portions of one larger RNA molecule, or they may be separate RNA molecules. Where the two strands are part of one larger molecule, and therefore are connected by an uninterrupted chain of nucleotides between the 3′-end of one strand and the 5′-end of the respective other strand forming the duplex structure, the connecting RNA chain is referred to as a “hairpin loop.” A hairpin loop can comprise at least one unpaired nucleotide. In some embodiments, the hairpin loop can comprise at at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, at least 20, at least 23 or more unpaired nucleotides or nucleotides not directed to the target site of the dsRNA. In some embodiments, the hairpin loop can be 10 or fewer nucleotides. In some embodiments, the hairpin loop can be 8 or fewer unpaired nucleotides. In some embodiments, the hairpin loop can be 4-10 unpaired nucleotides. In some embodiments, the hairpin loop can be 4-8 nucleotides. 
     In certain embodiment, the two strands of double-stranded oligomeric compound can be linked together. The two strands can be linked to each other at both ends, or at one end only. By linking at one end is meant that 5′-end of first strand is linked to the 3′-end of the second strand or 3′-end of first strand is linked to 5′-end of the second strand. When the two strands are linked to each other at both ends, 5′-end of first strand is linked to 3′-end of second strand and 3′-end of first strand is linked to 5′-end of second strand. The two strands can be linked together by an oligonucleotide linker including, but not limited to, (N)n; wherein N is independently a modified or unmodified nucleotide and n is 3-23. In some embodiments, n is 3-10, e.g., 3, 4, 5, 6, 7, 8, 9, or 10. In some embodiments, the oligonucleotide linker is selected from the group consisting of GNRA, (G)4, (U)4, and (dT)4, wherein N is a modified or unmodified nucleotide and R is a modified or unmodified purine nucleotide. Some of the nucleotides in the linker can be involved in base-pair interactions with other nucleotides in the linker. The two strands can also be linked together by a non-nucleosidic linker, e.g. a linker described herein. It will be appreciated by one of skill in the art that any oligonucleotide chemical modifications or variations describe herein can be used in the oligonucleotide linker. 
     Hairpin and dumbbell type oligomeric compounds will have a duplex region equal to or at least 14, 15, 15, 16, 17, 18, 19, 29, 21, 22, 23, 24, or 25 nucleotide pairs. The duplex region can be equal to or less than 200, 100, or 50, in length. In some embodiments, ranges for the duplex region are 15-30, 17 to 23, 19 to 23, and 19 to 21 nucleotides pairs in length. 
     The hairpin oligomeric compounds can have a single strand overhang or terminal unpaired region, in some embodiments at the 3′, and in some embodiments on the antisense side of the hairpin. In some embodiments, the overhangs are 1-4, more generally 2-3 nucleotides in length. The hairpin oligomeric compounds that can induce RNA interference are also referred to as “shRNA” herein. 
     Where the two substantially complementary strands of a dsRNA are comprised by separate RNA molecules, those molecules need not, but can be covalently connected. Where the two strands are connected covalently by means other than an uninterrupted chain of nucleotides between the 3′-end of one strand and the 5′-end of the respective other strand forming the duplex structure, the connecting structure is referred to as a “linker.” The RNA strands may have the same or a different number of nucleotides. The maximum number of base pairs is the number of nucleotides in the shortest strand of the dsRNA minus any overhangs that are present in the duplex. In addition to the duplex structure, an RNAi may comprise one or more nucleotide overhangs. 
     In one embodiment, an RNAi agent of the invention is a dsRNA, each strand of which is 24-30 nucleotides in length, that interacts with a target RNA sequence, e.g., an ACE2 mRNA sequence, to direct the cleavage of the target RNA. Without wishing to be bound by theory, long double stranded RNA introduced into cells is broken down into siRNA by a Type III endonuclease known as Dicer (Sharp et al. (2001)  Genes Dev.  15:485). Dicer, a ribonuclease-III-like enzyme, processes the dsRNA into 19-23 base pair short interfering RNAs with characteristic two base 3′ overhangs (Bernstein, et al., (2001)  Nature  409:363). The siRNAs are then incorporated into an RNA-induced silencing complex (RISC) where one or more helicases unwind the siRNA duplex, enabling the complementary antisense strand to guide target recognition (Nykanen, et al., (2001)  Cell  107:309). Upon binding to the appropriate target mRNA, one or more endonucleases within the RISC cleave the target to induce silencing (Elbashir, et al., (2001)  Genes Dev.  15:188). 
     In one embodiment, an RNAi agent of the invention is a dsRNA agent, each strand of which comprises 19-23 nucleotides that interacts with an ACE2 mRNA sequence to direct the cleavage of the target RNA. Without wishing to be bound by theory, long double stranded RNA introduced into cells is broken down into siRNA by a Type III endonuclease known as Dicer (Sharp et al. (2001)  Genes Dev.  15:485). Dicer, a ribonuclease-III-like enzyme, processes the dsRNA into 19-23 base pair short interfering RNAs with characteristic two base 3′ overhangs (Bernstein, et al., (2001)  Nature  409:363). The siRNAs are then incorporated into an RNA-induced silencing complex (RISC) where one or more helicases unwind the siRNA duplex, enabling the complementary antisense strand to guide target recognition (Nykanen, et al., (2001)  Cell  107:309). Upon binding to the appropriate target mRNA, one or more endonucleases within the RISC cleave the target to induce silencing (Elbashir, et al., (2001)  Genes Dev.  15:188). In one embodiment, an RNAi agent of the invention is a dsRNA of 24-30 nucleotides that interacts with an ACE2 mRNA sequence to direct the cleavage of the target RNA. 
     As used herein, the term “nucleotide overhang” refers to at least one unpaired nucleotide that protrudes from the duplex structure of a RNAi agent, e.g., a dsRNA. For example, when a 3′-end of one strand of a dsRNA extends beyond the 5′-end of the other strand, or vice versa, there is a nucleotide overhang. A dsRNA can comprise an overhang of at least one nucleotide; alternatively, the overhang can comprise at least two nucleotides, at least three nucleotides, at least four nucleotides, at least five nucleotides or more. A nucleotide overhang can comprise or consist of a nucleotide/nucleoside analog, including a deoxynucleotide/nucleoside. The overhang(s) can be on the sense strand, the antisense strand or any combination thereof. Furthermore, the nucleotide(s) of an overhang can be present on the 5′-end, 3′-end or both ends of either an antisense or sense strand of a dsRNA. 
     In one embodiment of the dsRNA, at least one strand comprises a 3′ overhang of at least 1 nucleotide. In another embodiment, at least one strand comprises a 3′ overhang of at least 2 nucleotides, e.g., 2, 3, 4, 5, 6, 7, 9, 10, 11, 12, 13, 14, or 15 nucleotides. In other embodiments, at least one strand of the RNAi agent comprises a 5′ overhang of at least 1 nucleotide. In certain embodiments, at least one strand comprises a 5′ overhang of at least 2 nucleotides, e.g., 2, 3, 4, 5, 6, 7, 9, 10, 11, 12, 13, 14, or 15 nucleotides. In still other embodiments, both the 3′ and the 5′ end of one strand of the RNAi agent comprise an overhang of at least 1 nucleotide. 
     In one embodiment, the antisense strand of a dsRNA has a 1-10 nucleotide, e.g., 0-3, 1-3, 2-4, 2-5, 4-10, 5-10, e.g., a 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 nucleotide, overhang at the 3′-end or the 5′-end. In one embodiment, the sense strand of a dsRNA has a 1-10 nucleotide, e.g., a 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 nucleotide, overhang at the 3′-end or the 5′-end. In another embodiment, one or more of the nucleotides in the overhang is replaced with a nucleoside thiophosphate. 
     In certain embodiments, the overhang on the sense strand or the antisense strand, or both, can include extended lengths longer than 10 nucleotides, e.g., 1-30 nucleotides, 2-30 nucleotides, 10-30 nucleotides, or 10-15 nucleotides in length. In certain embodiments, an extended overhang is on the sense strand of the duplex. In certain embodiments, an extended overhang is present on the 3′end of the sense strand of the duplex. In certain embodiments, an extended overhang is present on the 5′end of the sense strand of the duplex. In certain embodiments, an extended overhang is on the antisense strand of the duplex. In certain embodiments, an extended overhang is present on the 3′end of the antisense strand of the duplex. In certain embodiments, an extended overhang is present on the 5′end of the antisense strand of the duplex. In certain embodiments, one or more of the nucleotides in the overhang is replaced with a nucleoside thiophosphate. In certain embodiments, the overhang includes a self-complementary portion such that the overhang is capable of forming a hairpin structure that is stable under physiological conditions. 
     The terms “blunt” or “blunt ended” as used herein in reference to a dsRNA mean that there are no unpaired nucleotides or nucleotide analogs at a given terminal end of a dsRNA, i.e., no nucleotide overhang. One or both ends of a dsRNA can be blunt. Where both ends of a dsRNA are blunt, the dsRNA is said to be blunt ended. To be clear, a “blunt ended” dsRNA is a dsRNA that is blunt at both ends, i.e., no nucleotide overhang at either end of the molecule. Most often such a molecule will be double stranded over its entire length. 
     The term “antisense strand” or “guide strand” refers to the strand of an iRNA, e.g., a dsRNA, which includes a region that is substantially complementary to a target sequence, e.g., an ACE2 mRNA sequence. 
     As used herein, the term “region of complementarity” refers to the region on the antisense strand that is substantially complementary to a sequence, for example a target sequence, e.g., an ACE2 nucleotide sequence, as defined herein. Where the region of complementarity is not fully complementary to the target sequence, the mismatches can be in the internal or terminal regions of the molecule. Generally, the most tolerated mismatches are in the terminal regions, e.g., within 5, 4, 3, or 2 nucleotides of the 5′- or 3′-terminus of the RNAi agent. 
     In some embodiments, a double stranded RNA agent of the invention includes a nucleotide mismatch in the antisense strand. In some embodiments, the antisense strand of the double stranded RNA agent of the invention includes no more than 4 mismatches with the target mRNA, e.g., the antisense strand includes 4, 3, 2, 1, or 0 mismatches with the target mRNA. In some embodiments, the antisense strand double stranded RNA agent of the invention includes no more than 4 mismatches with the sense strand, e.g., the antisense strand includes 4, 3, 2, 1, or 0 mismatches with the sense strand. In some embodiments, a double stranded RNA agent of the invention includes a nucleotide mismatch in the sense strand. In some embodiments, the sense strand of the double stranded RNA agent of the invention includes no more than 4 mismatches with the antisense strand, e.g., the sense strand includes 4, 3, 2, 1, or 0 mismatches with the antisense strand. In some embodiments, the nucleotide mismatch is, for example, within 5, 4, 3 nucleotides from the 3′-end of the iRNA. In another embodiment, the nucleotide mismatch is, for example, in the 3′-terminal nucleotide of the iRNA agent. In some embodiments, the mismatch(s) is not in the seed region. 
     Thus, an RNAi agent as described herein can contain one or more mismatches to the target sequence. In one embodiment, a RNAi agent as described herein contains no more than 3 mismatches (i.e., 3, 2, 1, or 0 mismatches). In one embodiment, an RNAi agent as described herein contains no more than 2 mismatches. In one embodiment, an RNAi agent as described herein contains no more than 1 mismatch. In one embodiment, an RNAi agent as described herein contains 0 mismatches. In certain embodiments, if the antisense strand of the RNAi agent contains mismatches to the target sequence, the mismatch can optionally be restricted to be within the last 5 nucleotides from either the 5′- or 3′-end of the region of complementarity. For example, in such embodiments, for a 23 nucleotide RNAi agent, the strand which is complementary to a region of an ACE2 gene, generally does not contain any mismatch within the central 13 nucleotides. The methods described herein or methods known in the art can be used to determine whether an RNAi agent containing a mismatch to a target sequence is effective in inhibiting the expression of an ACE2 gene. Consideration of the efficacy of RNAi agents with mismatches in inhibiting expression of an ACE2 gene is important, especially if the particular region of complementarity in an ACE2 gene is known to vary. 
     The term “sense strand” or “passenger strand” as used herein, refers to the strand of a RNAi agent that includes a region that is substantially complementary to a region of the antisense strand as that term is defined herein. 
     As used herein, “substantially all of the nucleotides are modified” are largely but not wholly modified and can include not more than 5, 4, 3, 2, or 1 unmodified nucleotides. 
     As used herein, the term “cleavage region” refers to a region that is located immediately adjacent to the cleavage site. The cleavage site is the site on the target at which cleavage occurs. In some embodiments, the cleavage region comprises three bases on either end of, and immediately adjacent to, the cleavage site. In some embodiments, the cleavage region comprises two bases on either end of, and immediately adjacent to, the cleavage site. In some embodiments, the cleavage site specifically occurs at the site bound by nucleotides 10 and 11 of the antisense strand, and the cleavage region comprises nucleotides 11, 12 and 13. 
     As used herein, and unless otherwise indicated, the term “complementary,” when used to describe a first nucleotide sequence in relation to a second nucleotide sequence, refers to the ability of an oligonucleotide or polynucleotide comprising the first nucleotide sequence to hybridize and form a duplex structure under certain conditions with an oligonucleotide or polynucleotide comprising the second nucleotide sequence, as will be understood by the skilled person. Conditions, such as physiologically relevant conditions as can be encountered inside an organism, can apply. The skilled person will be able to determine the set of conditions most appropriate for a test of complementarity of two sequences in accordance with the ultimate application of the hybridized nucleotides. 
     Complementary sequences within a RNAi agent, e.g., within a dsRNA as described herein, include base-pairing of the oligonucleotide or polynucleotide comprising a first nucleotide sequence to an oligonucleotide or polynucleotide comprising a second nucleotide sequence over the entire length of one or both nucleotide sequences. Such sequences can be referred to as “fully complementary” with respect to each other herein. However, where a first sequence is referred to as “substantially complementary” with respect to a second sequence herein, the two sequences can be fully complementary, or they can form one or more, but generally not more than 5, 4, 3, or 2 mismatched base pairs upon hybridization for a duplex up to 30 base pairs, while retaining the ability to hybridize under the conditions most relevant to their ultimate application, e.g., inhibition of gene expression via a RISC pathway. However, where two oligonucleotides are designed to form, upon hybridization, one or more single stranded overhangs, such overhangs shall not be regarded as mismatches with regard to the determination of complementarity. For example, a dsRNA comprising one oligonucleotide 21 nucleotides in length and another oligonucleotide 23 nucleotides in length, wherein the longer oligonucleotide comprises a sequence of 21 nucleotides that is fully complementary to the shorter oligonucleotide, can yet be referred to as “fully complementary” for the purposes described herein. 
     “Complementary” sequences, as used herein, can also include, or be formed entirely from, non-Watson-Crick base pairs or base pairs formed from non-natural and modified nucleotides, in so far as the above requirements with respect to their ability to hybridize are fulfilled. Such non-Watson-Crick base pairs include, but are not limited to, G:U Wobble or Hoogstein base pairing. 
     The terms “complementary,” “fully complementary” and “substantially complementary” herein can be used with respect to the base matching between the sense strand and the antisense strand of a dsRNA, or between the antisense strand of a RNAi agent and a target sequence, as will be understood from the context of their use. 
     As used herein, a polynucleotide that is “substantially complementary to at least part of” a messenger RNA (mRNA) or target sequence refers to a polynucleotide that is substantially complementary to a contiguous portion of the mRNA of interest or target sequence (e.g., an mRNA encoding ACE2). For example, a polynucleotide is complementary to at least a part of an ACE2 RNA if the sequence is substantially complementary to a non-interrupted portion of an mRNA encoding ACE2. 
     Accordingly, in some embodiments, the antisense strand polynucleotides disclosed herein are fully complementary to the target ACE2 sequence. 
     In other embodiments, the antisense strand polynucleotides disclosed herein are substantially complementary to the target ACE2 sequence and comprise a contiguous nucleotide sequence which is at least about 80% complementary over its entire length to the equivalent region of the nucleotide sequence of SEQ ID NOs: 1-6 for ACE2, or a fragment of SEQ ID NOs: 1-6, such as about 85%, about 90%, or about 95% complementary. 
     In other embodiments, the antisense polynucleotides disclosed herein are substantially complementary to the target ACE2 sequence and comprise a contiguous nucleotide sequence which is at least about 80% complementary over its entire length to any one of the sense strand nucleotide sequences in any one of Tables 2-5, or a fragment of any one of the sense strand nucleotide sequences in any one of Tables 2-5, such as about 85%, about 90%, or about 95% complementary. 
     In one embodiment, an RNAi agent of the disclosure includes a sense strand that is substantially complementary to an antisense polynucleotide which, in turn, is the same as a target ACE2 sequence, and wherein the sense strand polynucleotide comprises a contiguous nucleotide sequence which is at least about 80% complementary over its entire length to the equivalent region of the nucleotide sequence of SEQ ID NOs: 7-12, or a fragment of any one of SEQ ID NOs: 7-12, such as about 85%, about 90%, or about 95% complementary. 
     In some embodiments, an iRNA of the invention includes a sense strand that is substantially complementary to an antisense polynucleotide which, in turn, is complementary to a target ACE2 sequence, and wherein the sense strand polynucleotide comprises a contiguous nucleotide sequence which is at least about 80% complementary over its entire length to any one of the antisense strand nucleotide sequences in any one of any one of Tables 2-5, or a fragment of any one of the antisense strand nucleotide sequences in any one of Tables 2-5, such as about 85%, about 90%, or about 95% complementary. 
     In some embodiments, the antisense polynucleotides disclosed herein are substantially complementary to a fragment of a target ACE2 sequence and comprise a contiguous nucleotide sequence which is at least 80% complementary over its entire length to a fragment of SEQ ID NO: 1 selected from the group of nucleotides 1695-1745, 1695-1735, 1695-1732, 1700-1745, 1700-1735, and 1704-1732 of SEQ ID NO: 1, such as about 85%, about 90%, about 95%, or fully complementary. 
     In certain embodiments, the sense and antisense strands are selected from any one of the chemically modified duplexes AAD-1230825, AD-1230843, AD-1230934. 
     In some embodiments, the double-stranded region of a double-stranded iRNA agent is equal to or at least, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 23, 24, 25, 26, 27, 28, 29, 30 or more nucleotide pairs in length. 
     In some embodiments, the antisense strand of a double-stranded iRNA agent is equal to or at least 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 23, 24, 25, 26, 27, 28, 29, or 30 nucleotides in length. 
     In some embodiments, the sense strand of a double-stranded iRNA agent is equal to or at least 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 23, 24, 25, 26, 27, 28, 29, or 30 nucleotides in length. 
     In one embodiment, the sense and antisense strands of the double-stranded iRNA agent are each independently 15 to 30 nucleotides in length. 
     In one embodiment, the sense and antisense strands of the double-stranded iRNA agent are each independently 19 to 25 nucleotides in length. 
     In one embodiment, the sense and antisense strands of the double-stranded iRNA agent are each independently 21 to 23 nucleotides in length. 
     In one embodiment, the sense strand of the iRNA agent is 21-nucleotides in length, and the antisense strand is 23-nucleotides in length, wherein the strands form a double-stranded region of 21 consecutive base pairs having a 2-nucleotide long single stranded overhangs at the 3′-end. 
     In one aspect of the invention, an agent for use in the methods and compositions of the invention is a single-stranded antisense nucleic acid molecule that inhibits a target mRNA via an antisense inhibition mechanism. The single-stranded antisense RNA molecule is complementary to a sequence within the target mRNA. The single-stranded antisense oligonucleotides can inhibit translation in a stoichiometric manner by base pairing to the mRNA and physically obstructing the translation machinery, see Dias, N. et al., (2002)  Mol Cancer Ther  1:347-355. The single-stranded antisense RNA molecule may be about 15 to about 30 nucleotides in length and have a sequence that is complementary to a target sequence. For example, the single-stranded antisense RNA molecule may comprise a sequence that is at least about 15, 16, 17, 18, 19, 20, or more contiguous nucleotides from any one of the antisense sequences described herein. 
     In one embodiment, at least partial suppression of the expression of an ACE2 gene, is assessed by a reduction of the amount of ACE2 mRNA which can be isolated from or detected in a first cell or group of cells in which an ACE2 gene is transcribed and which has or have been treated such that the expression of an ACE2 gene is inhibited, as compared to a second cell or group of cells substantially identical to the first cell or group of cells but which has or have not been so treated (control cells). The degree of inhibition may be expressed in terms of: 
     
       
         
           
             
               
                 
                   
                     ( 
                     
                       mRNA 
                       ⁢ 
                           
                       in 
                       ⁢ 
                           
                       control 
                       ⁢ 
                           
                       cells 
                     
                     ) 
                   
                   - 
                   
                     ( 
                     
                       mRNA 
                       ⁢ 
                           
                       in 
                       ⁢ 
                           
                       treated 
                       ⁢ 
                           
                       cells 
                     
                     ) 
                   
                 
                 
                   ( 
                   
                     mRNA 
                     ⁢ 
                         
                     in 
                     ⁢ 
                         
                     control 
                     ⁢ 
                         
                     cells 
                   
                   ) 
                 
               
               · 
               100 
             
             ⁢ 
             % 
           
         
       
     
     In one embodiment, inhibition of expression is determined by the dual luciferase method in Example 1 wherein the RNAi agent is present at 10 nM. 
     The phrase “contacting a cell with an RNAi agent,” such as a dsRNA, as used herein, includes contacting a cell by any possible means. Contacting a cell with an RNAi agent includes contacting a cell in vitro with the RNAi agent or contacting a cell in vivo with the RNAi agent. The contacting may be done directly or indirectly. Thus, for example, the RNAi agent may be put into physical contact with the cell by the individual performing the method, or alternatively, the RNAi agent may be put into a situation that will permit or cause it to subsequently come into contact with the cell. 
     Contacting a cell in vitro may be done, for example, by incubating the cell with the RNAi agent. Contacting a cell in vivo may be done, for example, via inhalation, intranasal administration, or intratracheal administration, by injecting the RNAi agent into or near the tissue where the cell is located, e.g., the pulmonary system, or by injecting the RNAi agent into another area, or to the bloodstream or the subcutaneous space, such that the agent will subsequently reach the tissue where the cell to be contacted is located. For example, the RNAi agent may contain or be coupled to a ligand, e.g., a lipophilic moiety or moieties as described below and further detailed, e.g., in PCT Publication No. WO 2019/217459, the entire contents of which is incorporated herein by reference, that directs or otherwise stabilizes the RNAi agent at a site of interest, e.g., the pulmonary system. In some embodiments, the RNAi agent may contain or be coupled to a ligand, e.g., one or more GalNAc derivatives as described below, that directs or otherwise stabilizes the RNAi agent at a site of interest, e.g., the liver. In other embodiments, the RNAi agent may contain or be coupled to a lipophilic moiety or moieties and one or more GalNAc derivatives. Combinations of in vitro and in vivo methods of contacting are also possible. For example, a cell may also be contacted in vitro with an RNAi agent and subsequently transplanted into a subject. 
     In one embodiment, contacting a cell with an RNAi agent includes “introducing” or “delivering the RNAi agent into the cell” by facilitating or effecting uptake or absorption into the cell. Absorption or uptake of a RNAi agent can occur through unaided diffusive or active cellular processes, or by auxiliary agents or devices. Introducing a RNAi agent into a cell may be in vitro or in vivo. For example, for in vivo introduction, a RNAi agent can be injected into a tissue site or administered systemically. In vitro introduction into a cell includes methods known in the art such as electroporation and lipofection. Further approaches are described herein below or are known in the art. 
     The term “lipophile” or “lipophilic moiety” broadly refers to any compound or chemical moiety having an affinity for lipids. One way to characterize the lipophilicity of the lipophilic moiety is by the octanol-water partition coefficient, logK ow , where K ow  is the ratio of a chemical&#39;s concentration in the octanol-phase to its concentration in the aqueous phase of a two-phase system at equilibrium. The octanol-water partition coefficient is a laboratory-measured property of a substance. However, it may also be predicted by using coefficients attributed to the structural components of a chemical which are calculated using first-principle or empirical methods (see, for example, Tetko et al.,  J. Chem. Inf. Comput. Sci.  41:1407-21 (2001), which is incorporated herein by reference in its entirety). It provides a thermodynamic measure of the tendency of the substance to prefer a non-aqueous or oily milieu rather than water (i.e. its hydrophilic/lipophilic balance). In principle, a chemical substance is lipophilic in character when its logK ow  exceeds 0. Typically, the lipophilic moiety possesses a logK ow  exceeding 1, exceeding 1.5, exceeding 2, exceeding 3, exceeding 4, exceeding 5, or exceeding 10. For instance, the logK ow  of 6-amino hexanol, for instance, is predicted to be approximately 0.7. Using the same method, the logK ow  of cholesteryl N-(hexan-6-ol) carbamate is predicted to be 10.7. 
     The lipophilicity of a molecule can change with respect to the functional group it carries. For instance, adding a hydroxyl group or amine group to the end of a lipophilic moiety can increase or decrease the partition coefficient (e.g., logK ow ) value of the lipophilic moiety. 
     Alternatively, the hydrophobicity of the double-stranded RNAi agent, conjugated to one or more lipophilic moieties, can be measured by its protein binding characteristics. For instance, in certain embodiments, the unbound fraction in the plasma protein binding assay of the double-stranded RNAi agent could be determined to positively correlate to the relative hydrophobicity of the double-stranded RNAi agent, which could then positively correlate to the silencing activity of the double-stranded RNAi agent. 
     In one embodiment, the plasma protein binding assay determined is an electrophoretic mobility shift assay (EMSA) using human serum albumin protein. An exemplary protocol of this binding assay is illustrated in detail in, e.g., PCT Publication No. WO 2019/217459. The hydrophobicity of the double-stranded RNAi agent, measured by fraction of unbound siRNA in the binding assay, exceeds 0.15, exceeds 0.2, exceeds 0.25, exceeds 0.3, exceeds 0.35, exceeds 0.4, exceeds 0.45, or exceeds 0.5 for an enhanced in vivo delivery of siRNA. 
     Accordingly, conjugating the lipophilic moieties to the internal position(s) of the double-stranded RNAi agent provides optimal hydrophobicity for the enhanced in vivo delivery of siRNA. 
     The term “lipid nanoparticle” or “LNP” is a vesicle comprising a lipid layer encapsulating a pharmaceutically active molecule, such as a nucleic acid molecule, e.g., a rNAi agent or a plasmid from which a RNAi agent is transcribed. LNPs are described in, for example, U.S. Pat. Nos. 6,858,225, 6,815,432, 8,158,601, and 8,058,069, the entire contents of which are hereby incorporated herein by reference. 
     As used herein, a “subject” is an animal, such as a mammal, including a primate (such as a human, a non-human primate, e.g., a monkey, and a chimpanzee), or a non-primate (such as a a cow, a pig, a horse, a goat, a rabbit, a sheep, a hamster, a guinea pig, a cat, a dog, a rat, or a mouse), or a bird that expresses the target gene, either endogenously or heterologously. In a preferred embodiment, the subject is a human, such as a human being treated or assessed for a disease, disorder, or condition that would benefit from reduction in ACE2 expression; a human at risk for a disease, disorder, or condition that would benefit from reduction in ACE2 expression; a human having a disease, disorder, or condition that would benefit from reduction in ACE2 expression; or human being treated for a disease, disorder, or condition that would benefit from reduction in ACE2 expression as described herein. In some embodiments, the subject is a female human. In other embodiments, the subject is a male human. In one embodiment, the subject is an adult subject. In another embodiment, the subject is a pediatric subject. 
     As used herein, the terms “treating” or “treatment” refer to a beneficial or desired result including, but not limited to, alleviation or amelioration of one or more signs or symptoms associated with ACE2 expression or ACE2 protein production, e.g., an ACE2-associated disease, e.g., a coronavirus-associated disease. Treatment also includes a reduction of one or more sign or symptoms associated with unwanted ACE2 expression; diminishing the extent of unwanted ACE2 activation or stabilization; amelioration or palliation of unwanted ACE2 activation or stabilization. “Treatment” can also mean prolonging survival as compared to expected survival in the absence of treatment. 
     The term “lower” in the context of the level of ACE2 in a subject or a disease marker or symptom refers to a statistically significant decrease in such level. The decrease can be, for example, at least 10%, 15%, 20%, 25%, 30%, %, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or more. In certain embodiments, a decrease is at least 20%. In certain embodiments, the decrease is at least 50% in a disease marker, e.g., protein or gene expression level. “Lower” in the context of the level of ACE2 in a subject is preferably down to a level accepted as within the range of normal for an individual without such disorder. In certain embodiments, the expression of the target is normalized, i.e., decreased towards or to a level accepted as within the range of normal for an individual without such disorder, e.g., viral load, blood oxygen level, white blood cell count, kidney function, liver function. As used here, “lower” in a subject can refer to lowering of gene expression or protein production in a cell in a subject does not require lowering of expression in all cells or tissues of a subject. For example, as used herein, lowering in a subject can include lowering of gene expression or protein production or viral replication in a subject. 
     The term “lower” can also be used in association with normalizing a symptom of a disease or condition, i.e. decreasing the difference between a level in a subject suffering from an ACE2-associated disease towards or to a level in a normal subject not suffering from an ACE2-associated disease. As used herein, if a disease is associated with an elevated value for a symptom, “normal” is considered to be the upper limit of normal. If a disease is associated with a decreased value for a symptom, “normal” is considered to be the lower limit of normal. 
     As used herein, “prevention” or “preventing,” when used in reference to a disease, disorder, or condition thereof, that would benefit from a reduction in expression of an ACE2 gene or production of an ACE2 protein, refers to a reduction in the likelihood that a subject will develop a symptom associated with such a disease, disorder, or condition, e.g., a symptom of an ACE2-associated disease, such as COVID-19. The failure to develop a disease, disorder, or condition, or the reduction in the development of a symptom associated with such a disease, disorder, or condition, e.g., pneumonia (e.g., by at least about 10% on a clinically accepted scale for that disease or disorder), or the exhibition of delayed symptoms (e.g., delayed by days, weeks, months or years) is considered effective prevention. 
     As used herein, the term “ACE2-associated disease,” is a disease or disorder that would benefit from reduction in the expression or activity of ACE2. Such ACE2-associated diseases include a coronavirus-associated disease. The term “coronavirus-associated disease,” is a disease or disorder that is caused by, or associated with a coronavirus infection, coronavirus genome expression or coronavirus protein production. The term “coronavirus-associated disease” includes a disease, disorder or condition that would benefit from a decrease in coronavirus S protein priming, viral genome expression, cellular (viral) entry, viral replication, or viral protein activity. 
     Non-limiting examples of coronavirus-associated diseases include, for example, disease or disorders caused by infection with human coronavirus 229E (HCoV-229E), human coronavirus NL63 (HCoV-NL63), human coronavirus OC43 (HCoV-OC43), human coronavirus HKU1 (HCoV-HKU1), severe acute respiratory syndrome coronavirus (SARS), the Middle East respiratory syndrome coronavirus (MERS), and severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2 or COVID-19). The symptoms for a coronavirus-associated disease depend on the type of coronavirus and how serious the infection is. Patients with a mild to moderate upper-respiratory infection may develop symptoms such as runny nose, sneezing, headache, cough, sore throat, fever, or short of breath. In more severe cases, coronavirus infection can cause pneumonia, severe acute respiratory syndrome, kidney failure and even death. Further details regarding signs and symptoms of the various diseases or conditions are provided herein and are well known in the art. 
     “Therapeutically effective amount,” as used herein, is intended to include the amount of an RNAi agent that, when administered to a subject having a coronavirus-associated disease, is sufficient to effect treatment of the disease (e.g., by diminishing, ameliorating, or maintaining the existing disease or one or more symptoms of disease). The “therapeutically effective amount” may vary depending on the RNAi agent, how the agent is administered, the disease and its severity and the history, age, weight, family history, genetic makeup, the types of preceding or concomitant treatments, if any, and other individual characteristics of the subject to be treated. 
     “Prophylactically effective amount,” as used herein, is intended to include the amount of a RNAi agent that, when administered to a subject having an ACE2-associated disorder, e.g., a coronavirus-associated disorder, such as COVID-19, is sufficient to prevent or ameliorate the disease or one or more symptoms of the disease. Ameliorating the disease includes slowing the course of the disease or reducing the severity of later-developing disease. The “prophylactically effective amount” may vary depending on the RNAi agent, how the agent is administered, the degree of risk of disease, and the history, age, weight, family history, genetic makeup, the types of preceding or concomitant treatments, if any, and other individual characteristics of the patient to be treated. 
     A “therapeutically-effective amount” or “prophylactically effective amount” also includes an amount of a RNAi agent that produces some desired local or systemic effect at a reasonable benefit/risk ratio applicable to any treatment. A RNAi agent employed in the methods of the present disclosure may be administered in a sufficient amount to produce a reasonable benefit/risk ratio applicable to such treatment. 
     The phrase “pharmaceutically acceptable” is employed herein to refer to those compounds, materials, compositions, or dosage forms which are, within the scope of sound medical judgment, suitable for use in contact with the tissues of human subjects and animal subjects without excessive toxicity, irritation, allergic response, or other problem or complication, commensurate with a reasonable benefit/risk ratio. 
     The phrase “pharmaceutically-acceptable carrier” as used herein means a pharmaceutically-acceptable material, composition or vehicle, such as a liquid or solid filler, diluent, excipient, manufacturing aid (e.g., lubricant, talc magnesium, calcium or zinc stearate, or steric acid), or solvent encapsulating material, involved in carrying or transporting the subject compound from one organ, or portion of the body, to another organ, or portion of the body. Each carrier must be “acceptable” in the sense of being compatible with the other ingredients of the formulation and not injurious to the subject being treated. Some examples of materials which can serve as pharmaceutically-acceptable carriers include: (1) sugars, such as lactose, glucose and sucrose; (2) starches, such as corn starch and potato starch; (3) cellulose, and its derivatives, such as sodium carboxymethyl cellulose, ethyl cellulose and cellulose acetate; (4) powdered tragacanth; (5) malt; (6) gelatin; (7) lubricating agents, such as magnesium state, sodium lauryl sulfate and talc; (8) excipients, such as cocoa butter and suppository waxes; (9) oils, such as peanut oil, cottonseed oil, safflower oil, sesame oil, olive oil, corn oil and soybean oil; (10) glycols, such as propylene glycol; (11) polyols, such as glycerin, sorbitol, mannitol and polyethylene glycol; (12) esters, such as ethyl oleate and ethyl laurate; (13) agar; (14) buffering agents, such as magnesium hydroxide and aluminum hydroxide; (15) alginic acid; (16) pyrogen-free water; (17) isotonic saline; (18) Ringer&#39;s solution; (19) ethyl alcohol; (20) pH buffered solutions; (21) polyesters, polycarbonates or polyanhydrides; (22) bulking agents, such as polypeptides and amino acids (23) serum component, such as serum albumin, HDL and LDL; and (22) other non-toxic compatible substances employed in pharmaceutical formulations. 
     The term “sample,” as used herein, includes a collection of similar fluids, cells, or tissues isolated from a subject, as well as fluids, cells, or tissues present within a subject. Examples of biological fluids include blood, serum and serosal fluids, plasma, bronchial fluids, sputum, cerebrospinal fluid, ocular fluids, lymph, urine, saliva, and the like. Tissue samples may include samples from tissues, organs or localized regions. For example, samples may be derived from particular organs, parts of organs, or fluids or cells within those organs. In certain embodiments, samples may be derived from a nasal swab. In certain embodiments, samples may be derived from a throat swab/ In certain embodiments, samples may be derived from the lung, or certain types of cells in the lung. In some embodiments, the samples may be derived from the bronchioles. In some embodiments, the samples may be derived from the bronchus. In some embodiments, the samples may be derived from the alveoli. In other embodiments, a “sample derived from a subject” refers to liver tissue (or subcomponents thereof) derived from the subject. In some embodiments, a “sample derived from a subject” refers to blood drawn from the subject or plasma or serum derived therefrom. In further embodiments, a “sample derived from a subject” refers to pulmonary tissue (or subcomponents thereof) derived from the subject. 
     II. RNAI AGENTS OF THE DISCLOSURE 
     Described herein are RNAi agents which inhibit the expression of an ACE2 gene. In one embodiment, the RNAi agent includes double stranded ribonucleic acid (dsRNA) molecules for inhibiting the expression of an ACE2 gene in a cell, such as a cell within a subject, e.g., a mammal, such as a human, e.g., a subject having an ACE2-associated disorder, e.g., a coronavirus-associated disorder, e.g., a subject having a coronavirus infection, e.g., a subject having Severe Acute Respiratory Syndrome 2 (SARS-CoV-2; COVID-19), Severe Acute Respiratory Syndrome (SARS-CoV), or Middle East Respiratory Syndrome (MERS-CoV). The dsRNA includes an antisense strand having a region of complementarity which is complementary to at least a part of a target RNA, e.g., an mRNA formed in the expression of an ACE2 gene. The region of complementarity is about 15-30 nucleotides or less in length. Upon contact with a cell expressing the ACE2 gene, the RNAi agent inhibits the expression of the ACE2 gene (e.g., a human gene, a primate gene, a non-primate gene) by at least 50% as assayed by, for example, a PCR or branched DNA (bDNA)-based method, or by a protein-based method, such as by immunofluorescence analysis, using, for example, western blotting or flowcytometric techniques. In preferred embodiments, inhibition of expression is by at least 50% as assayed by the Dual-Glo lucifierase assay in Example 1 where the siRNA is at a 10 nM concentration. 
     A dsRNA includes two RNA strands that are complementary and hybridize to form a duplex structure under conditions in which the dsRNA will be used. One strand of a dsRNA (the antisense strand) includes a region of complementarity that is substantially complementary, and generally fully complementary, to a target sequence. For example, the target sequence can be derived from the sequence of an mRNA formed during the expression of an ACE2 gene. The other strand (the sense strand) includes a region that is complementary to the antisense strand, such that the two strands hybridize and form a duplex structure when combined under suitable conditions. As described elsewhere herein and as known in the art, the complementary sequences of a dsRNA can also be contained as self-complementary regions of a single nucleic acid molecule, as opposed to being on separate oligonucleotides. 
     Generally, the duplex structure is 15 to 30 base pairs in length, e.g., 15-29, 15-28, 15-27, 15-26, 15-25, 15-24, 15-23, 15-22, 15-21, 15-20, 15-19, 15-18, 15-17, 18-30, 18-29, 18-28, 18-27, 18-26, 18-25, 18-24, 18-23, 18-22, 18-21, 18-20, 19-30, 19-29, 19-28, 19-27, 19-26, 19-25, 19-24, 19-23, 19-22, 19-21, 19-20, 20-30, 20-29, 20-28, 20-27, 20-26, 20-25, 20-24,20-23, 20-22, 20-21, 21-30, 21-29, 21-28, 21-27, 21-26, 21-25, 21-24, 21-23, or 21-22 base pairs in length. In certain preferred embodiments, the duplex structure is 18 to 25 base pairs in length, e.g., 18-25, 18-24, 18-23, 18-22, 18-21, 18-20, 19-25, 19-24, 19-23, 19-22, 19-21, 19-20, 20-25, 20-24,20-23, 20-22, 20-21, 21-25, 21-24, 21-23, 21-22, 22-25, 22-24, 22-23, 23-25, 23-24 or 24-25 base pairs in length, for example, 19-21 basepairs in length. Ranges and lengths intermediate to the above recited ranges and lengths are also contemplated to be part of the disclosure. 
     Similarly, the region of complementarity to the target sequence is 15 to 30 nucleotides in length, e.g., 15-29, 15-28, 15-27, 15-26, 15-25, 15-24, 15-23, 15-22, 15-21, 15-20, 15-19, 15-18, 15-17, 18-30, 18-29, 18-28, 18-27, 18-26, 18-25, 18-24, 18-23, 18-22, 18-21, 18-20, 19-30, 19-29, 19-28, 19-27, 19-26, 19-25, 19-24, 19-23, 19-22, 19-21, 19-20, 20-30, 20-29, 20-28, 20-27, 20-26, 20-25, 20-24,20-23, 20-22, 20-21, 21-30, 21-29, 21-28, 21-27, 21-26, 21-25, 21-24, 21-23, or 21-22 nucleotides in length, for example 19-23 nucleotides in length or 21-23 nucleotides in length. Ranges and lengths intermediate to the above recited ranges and lengths are also contemplated to be part of the disclosure. 
     In some embodiments, the dsRNA is 15 to 23 nucleotides in length, or 25 to 30 nucleotides in length. In general, the dsRNA is long enough to serve as a substrate for the Dicer enzyme. For example, it is well known in the art that dsRNAs longer than about 21-23 nucleotides can serve as substrates for Dicer. As the ordinarily skilled person will also recognize, the region of an RNA targeted for cleavage will most often be part of a larger RNA molecule, often an mRNA molecule. Where relevant, a “part” of an mRNA target is a contiguous sequence of an mRNA target of sufficient length to allow it to be a substrate for RNAi-directed cleavage (i.e., cleavage through a RISC pathway). 
     One of skill in the art will also recognize that the duplex region is a primary functional portion of a dsRNA, e.g., a duplex region of about 15 to 36 base pairs, e.g., 15-36, 15-35, 15-34, 15-33, 15-32, 15-31, 15-30, 15-29, 15-28, 15-27, 15-26, 15-25, 15-24, 15-23, 15-22, 15-21, 15-20, 15-19, 15-18, 15-17, 18-30, 18-29, 18-28, 18-27, 18-26, 18-25, 18-24, 18-23, 18-22, 18-21, 18-20, 19-30, 19-29, 19-28, 19-27, 19-26, 19-25, 19-24, 19-23, 19-22, 19-21, 19-20, 20-30, 20-29, 20-28, 20-27, 20-26, 20-25, 20-24,20-23, 20-22, 20-21, 21-30, 21-29, 21-28, 21-27, 21-26, 21-25, 21-24, 21-23, or 21-22 base pairs, for example, 19-21 base pairs. Thus, in one embodiment, to the extent that it becomes processed to a functional duplex, of e.g., 15-30 base pairs, that targets a desired RNA for cleavage, an RNA molecule or complex of RNA molecules having a duplex region greater than 30 base pairs is a dsRNA. Thus, an ordinarily skilled artisan will recognize that in one embodiment, a miRNA is a dsRNA. In another embodiment, a dsRNA is not a naturally occurring miRNA. In another embodiment, a RNAi agent useful to target ACE2 expression is not generated in the target cell by cleavage of a larger dsRNA. 
     A dsRNA as described herein can further include one or more single-stranded nucleotide overhangs e.g., 1, 2, 3, or 4 nucleotides. A nucleotide overhang can comprise or consist of a nucleotide/nucleoside analog, including a deoxynucleotide/nucleoside. The overhang(s) can be on the sense strand, the antisense strand or any combination thereof. Furthermore, the nucleotide(s) of an overhang can be present on the 5′-end, 3′-end or both ends of either an antisense or sense strand of a dsRNA. In certain embodiments, longer, extended overhangs are possible. 
     A dsRNA can be synthesized by standard methods known in the art as further discussed below, e.g., by use of an automated DNA synthesizer, such as are commercially available from, for example, Biosearch, Applied Biosystems, Inc. 
     iRNA compounds of the invention may be prepared using a two-step procedure. First, the individual strands of the double stranded RNA molecule are prepared separately. Then, the component strands are annealed. The individual strands of the siRNA compound can be prepared using solution-phase or solid-phase organic synthesis or both. Organic synthesis offers the advantage that the oligonucleotide strands comprising unnatural or modified nucleotides can be easily prepared. Single-stranded oligonucleotides of the invention can be prepared using solution-phase or solid-phase organic synthesis or both. 
     An siRNA can be produced, e.g., in bulk, by a variety of methods. Exemplary methods include: organic synthesis and RNA cleavage, e.g., in vitro cleavage. 
     An siRNA can be made by separately synthesizing a single stranded RNA molecule, or each respective strand of a double-stranded RNA molecule, after which the component strands can then be annealed. 
     A large bioreactor, e.g., the OligoPilot II from Pharmacia Biotec AB (Uppsala Sweden), can be used to produce a large amount of a particular RNA strand for a given siRNA. The OligoPilotII reactor can efficiently couple a nucleotide using only a 1.5 molar excess of a phosphoramidite nucleotide. To make an RNA strand, ribonucleotides amidites are used. Standard cycles of monomer addition can be used to synthesize the 21 to 23 nucleotide strand for the siRNA. Typically, the two complementary strands are produced separately and then annealed, e.g., after release from the solid support and deprotection. 
     Organic synthesis can be used to produce a discrete siRNA species. The complementary of the species to an ACE2 gene can be precisely specified. For example, the species may be complementary to a region that includes a polymorphism, e.g., a single nucleotide polymorphism. Further the location of the polymorphism can be precisely defined. In some embodiments, the polymorphism is located in an internal region, e.g., at least 4, 5, 7, or 9 nucleotides from one or both of the termini. 
     In one embodiment, RNA generated is carefully purified to remove endsiRNA is cleaved in vitro into siRNAs, for example, using a Dicer or comparable RNAse III-based activity. For example, the dsiRNA can be incubated in an in vitro extract from  Drosophila  or using purified components, e.g., a purified RNAse or RISC complex (RNA-induced silencing complex). See, e.g., Ketting et al.  Genes Dev  2001 Oct. 15; 15(20):2654-9 and Hammond  Science  2001 Aug. 10; 293(5532):1146-50. 
     dsiRNA cleavage generally produces a plurality of siRNA species, each being a particular 21 to 23 nucleotide fragment of a source dsiRNA molecule. For example, siRNAs that include sequences complementary to overlapping regions and adjacent regions of a source dsiRNA molecule may be present. 
     Regardless of the method of synthesis, the siRNA preparation can be prepared in a solution (e.g., an aqueous or organic solution) that is appropriate for formulation. For example, the siRNA preparation can be precipitated and redissolved in pure double-distilled water, and lyophilized. The dried siRNA can then be resuspended in a solution appropriate for the intended formulation process. 
     In one aspect, a dsRNA of the disclosure includes at least two nucleotide sequences, a sense sequence and an antisense sequence. The sense strand sequence for ACE2 may be selected from the group of sequences provided in any one of Tables 2-5, and the corresponding nucleotide sequence of the antisense strand of the sense strand may be selected from the group of sequences of any one of Tables 2-5. In this aspect, one of the two sequences is complementary to the other of the two sequences, with one of the sequences being substantially complementary to a sequence of an mRNA generated in the expression of an ACE2 gene. As such, in this aspect, a dsRNA will include two oligonucleotides, where one oligonucleotide is described as the sense strand (passenger strand) in any one of Tables 2-5, and the second oligonucleotide is described as the corresponding antisense strand (guide strand) of the sense strand in any one of Tables 2-5 for ACE2. 
     In certain embodiments, the sense or antisense strand is selected from the sense or antisense strand of any one of duplexes AD-1230825, AD-1230843, or AD-1230934. 
     In one embodiment, the substantially complementary sequences of the dsRNA are contained on separate oligonucleotides. In another embodiment, the substantially complementary sequences of the dsRNA are contained on a single oligonucleotide. 
     It will be understood that, although the sequences provided herein are described as modified or conjugated sequences, the RNA of the RNAi agent of the disclosure e.g., a dsRNA of the disclosure, may comprise any one of the sequences set forth in any one of Tables 2-5 that is unmodified, un-conjugated, or modified or conjugated differently than described therein. One or more lipophilic ligands or one or more GalNAc ligands can be included in any of the positions of the RNAi agents provided in the instant application. 
     The skilled person is well aware that dsRNAs having a duplex structure of about 20 to 23 base pairs, e.g., 21, base pairs have been hailed as particularly effective in inducing RNA interference (Elbashir et al., (2001)  EMBO J.,  20:6877-6888). However, others have found that shorter or longer RNA duplex structures can also be effective (Chu and Rana (2007)  RNA  14:1714-1719; Kim et al. (2005)  Nat Biotech  23:222-226). In the embodiments described above, by virtue of the nature of the oligonucleotide sequences provided herein, dsRNAs described herein can include at least one strand of a length of minimally 21 nucleotides. It can be reasonably expected that shorter duplexes minus only a few nucleotides on one or both ends can be similarly effective as compared to the dsRNAs described above. Hence, dsRNAs having a sequence of at least 15, 16, 17, 18, 19, 20, or more contiguous nucleotides derived from one of the sequences provided herein, and differing in their ability to inhibit the expression of an ACE2 gene by not more than 10, 15, 20, 25, or 30% inhibition from a dsRNA comprising the full sequence using the in vitro assay with Cos7 and a 10 nM concentration of the RNA agent and the PCR assay as provided in the examples herein, are contemplated to be within the scope of the present disclosure. 
     In addition, the RNAs described herein identify a site(s) in an ACE2 transcript that is susceptible to RISC-mediated cleavage. As such, the present disclosure further features RNAi agents that target within this site(s). As used herein, a RNAi agent is said to target within a particular site of an RNA transcript if the RNAi agent promotes cleavage of the transcript anywhere within that particular site. Such a RNAi agent will generally include at least about 15 contiguous nucleotides, preferably at least 19 nucleotides, from one of the sequences provided herein coupled to additional nucleotide sequences taken from the region contiguous to the selected sequence in an ACE2 gene. 
     An RNAi agent as described herein can contain one or more mismatches to the target sequence. In one embodiment, an RNAi agent as described herein contains no more than 3 mismatches (i.e., 3, 2, 1, or 0 mismatches). In one embodiment, an RNAi agent as described herein contains no more than 2 mismatches. In one embodiment, an RNAi agent as described herein contains no more than 1 mismatch. In one embodiment, an RNAi agent as described herein contains 0 mismatches. In certain embodiments, if the antisense strand of the RNAi agent contains mismatches to the target sequence, the mismatch can optionally be restricted to be within the last 5 nucleotides from either the 5′- or 3′-end of the region of complementarity. For example, in such embodiments, for a 23 nucleotide RNAi agent, the strand which is complementary to a region of an ACE2 gene generally does not contain any mismatch within the central 13 nucleotides. The methods described herein or methods known in the art can be used to determine whether an RNAi agent containing a mismatch to a target sequence is effective in inhibiting the expression of an ACE2 gene. Consideration of the efficacy of RNAi agents with mismatches in inhibiting expression of an ACE2 gene is important, especially if the particular region of complementarity in an ACE2 gene is known to mutate. 
     III. MODIFIED RNAI AGENTS OF THE DISCLOSURE 
     In one embodiment, the RNA of the RNAi agent of the disclosure e.g., a dsRNA, is unmodified, and does not comprise, e.g., chemical modifications or conjugations known in the art and described herein. In preferred embodiments, the RNA of an RNAi agent of the disclosure, e.g., a dsRNA, is chemically modified to enhance stability or other beneficial characteristics. In certain embodiments of the disclosure, substantially all of the nucleotides of an RNAi agent of the disclosure are modified. In other embodiments of the disclosure, all of the nucleotides of an RNAi agent of the disclosure are modified. RNAi agents of the disclosure in which “substantially all of the nucleotides are modified” are largely but not wholly modified and can include not more than 5, 4, 3, 2, or 1 unmodified nucleotides. In still other embodiments of the disclosure, RNAi agents of the disclosure can include not more than 5, 4, 3, 2 or 1 modified nucleotides. 
     The nucleic acids featured in the disclosure can be synthesized or modified by methods well established in the art, such as those described in “Current protocols in nucleic acid chemistry,” Beaucage, S. L. et al. (Edrs.), John Wiley &amp; Sons, Inc., New York, N.Y., USA, which is hereby incorporated herein by reference. Modifications include, for example, end modifications, e.g., 5′-end modifications (phosphorylation, conjugation, inverted linkages) or 3′-end modifications (conjugation, DNA nucleotides, inverted linkages, etc.); base modifications, e.g., replacement with stabilizing bases, destabilizing bases, or bases that base pair with an expanded repertoire of partners, removal of bases (abasic nucleotides), or conjugated bases; sugar modifications (e.g., at the 2′-position or 4′-position) or replacement of the sugar; or backbone modifications, including modification or replacement of the phosphodiester linkages. Specific examples of RNAi agents useful in the embodiments described herein include, but are not limited to, RNAs containing modified backbones or no natural internucleoside linkages. RNAs having modified backbones include, among others, those that do not have a phosphorus atom in the backbone. For the purposes of this specification, and as sometimes referenced in the art, modified RNAs that do not have a phosphorus atom in their internucleoside backbone can also be considered to be oligonucleosides. In some embodiments, a modified RNAi agent will have a phosphorus atom in its internucleoside backbone. 
     Modified RNA backbones include, for example, phosphorothioates, chiral phosphorothioates, phosphorodithioates, phosphotriesters, aminoalkylphosphotriesters, methyl and other alkyl phosphonates including 3′-alkylene phosphonates and chiral phosphonates, phosphinates, phosphoramidates including 3′-amino phosphoramidate and aminoalkylphosphoramidates, thionophosphoramidates, thionoalkylphosphonates, thionoalkylphosphotriesters, and boranophosphates having normal 3′-5′ linkages, 2′-5′-linked analogs of these, and those having inverted polarity wherein the adjacent pairs of nucleoside units are linked 3′-5′ to 5′-3′ or 2′-5′ to 5′-2′. Various salts, mixed salts and free acid forms are also included. In some embodiments of the invention, the dsRNA agents of the invention are in a free acid form. In other embodiments of the invention, the dsRNA agents of the invention are in a salt form. In one embodiment, the dsRNA agents of the invention are in a sodium salt form. In certain embodiments, when the dsRNA agents of the invention are in the sodium salt form, sodium ions are present in the agent as counterions for substantially all of the phosphodiester or phosphorothiotate groups present in the agent. Agents in which substantially all of the phosphodiester or phosphorothioate linkages have a sodium counterion include not more than 5, 4, 3, 2, or 1 phosphodiester or phosphorothioate linkages without a sodium counterion. In some embodiments, when the dsRNA agents of the invention are in the sodium salt form, sodium ions are present in the agent as counterions for all of the phosphodiester or phosphorothiotate groups present in the agent. 
     Representative U.S. patents that teach the preparation of the above phosphorus-containing linkages include, but are not limited to, U.S. Pat. Nos. 3,687,808; 4,469,863; 4,476,301; 5,023,243; 5,177,195; 5,188,897; 5,264,423; 5,276,019; 5,278,302; 5,286,717; 5,321,131; 5,399,676; 5,405,939; 5,453,496; 5,455,233; 5,466,677; 5,476,925; 5,519,126; 5,536,821; 5,541,316; 5,550,111; 5,563,253; 5,571,799; 5,587,361; 5,625,050; 6,028,188; 6,124,445; 6,160,109; 6,169,170; 6,172,209; 6,239,265; 6,277,603; 6,326,199; 6,346,614; 6,444,423; 6,531,590; 6,534,639; 6,608,035; 6,683,167; 6,858,715; 6,867,294; 6,878,805; 7,015,315; 7,041,816; 7,273,933; 7,321,029; and U.S. Pat. RE39464, the entire contents of each of which are hereby incorporated herein by reference. 
     Modified RNA backbones that do not include a phosphorus atom therein have backbones that are formed by short chain alkyl or cycloalkyl internucleoside linkages, mixed heteroatoms and alkyl or cycloalkyl internucleoside linkages, or one or more short chain heteroatomic or heterocyclic internucleoside linkages. These include those having morpholino linkages (formed in part from the sugar portion of a nucleoside); siloxane backbones; sulfide, sulfoxide and sulfone backbones; formacetyl and thioformacetyl backbones; methylene formacetyl and thioformacetyl backbones; alkene containing backbones; sulfamate backbones; methyleneimino and methylenehydrazino backbones; sulfonate and sulfonamide backbones; amide backbones; and others having mixed N, O, S and CH 2  component parts. 
     Representative U.S. patents that teach the preparation of the above oligonucleosides include, but are not limited to, U.S. Pat. Nos. 5,034,506; 5,166,315; 5,185,444; 5,214,134; 5,216,141; 5,235,033; 5,64,562; 5,264,564; 5,405,938; 5,434,257; 5,466,677; 5,470,967; 5,489,677; 5,541,307; 5,561,225; 5,596,086; 5,602,240; 5,608,046; 5,610,289; 5,618,704; 5,623,070; 5,663,312; 5,633,360; 5,677,437; and, 5,677,439, the entire contents of each of which are hereby incorporated herein by reference. 
     In other embodiments, suitable RNA mimetics are contemplated for use in RNAi agents, in which both the sugar and the internucleoside linkage, i.e., the backbone, of the nucleotide units are replaced with novel groups. The base units are maintained for hybridization with an appropriate nucleic acid target compound. One such oligomeric compound, an RNA mimetic that has been shown to have excellent hybridization properties, is referred to as a peptide nucleic acid (PNA). In PNA compounds, the sugar backbone of an RNA is replaced with an amide containing backbone, in particular an aminoethylglycine backbone. The nucleobases are retained and are bound directly or indirectly to aza nitrogen atoms of the amide portion of the backbone. Representative U.S. patents that teach the preparation of PNA compounds include, but are not limited to, U.S. Pat. Nos. 5,539,082; 5,714,331; and 5,719,262, the entire contents of each of which are hereby incorporated herein by reference. Additional PNA compounds suitable for use in the RNAi agents of the disclosure are described in, for example, in Nielsen et al.,  Science,  1991, 254, 1497-1500. 
     Some embodiments featured in the disclosure include RNAs with phosphorothioate backbones and oligonucleosides with heteroatom backbones, and in particular —CH 2 —NH—CH 2 —, —CH 2 —N(CH 3 )—O—CH 2 — [known as a methylene (methylimino) or MMI backbone], —CH 2 —O—N(CH 3 )—CH 2 —, —CH 2 —N(CH 3 )—N(CH 3 )—CH 2 — and —N(CH 3 )—CH 2 —CH 2 — [wherein the native phosphodiester backbone is represented as —O—P—O—CH 2 —] of the above-referenced U.S. Pat. No. 5,489,677, and the amide backbones of the above-referenced U.S. Pat. No. 5,602,240. In some embodiments, the RNAs featured herein have morpholino backbone structures of the above-referenced U.S. Pat. No. 5,034,506. 
     Modified RNAs can also contain one or more substituted sugar moieties. The RNAi agents, e.g., dsRNAs, featured herein can include one of the following at the 2′-position: OH; F; O-, S-, or N-alkyl; O-, S-, or N-alkenyl; O—, S- or N-alkynyl; or O-alkyl-O-alkyl, wherein the alkyl, alkenyl and alkynyl can be substituted or unsubstituted C 1  to C 10  alkyl or C 2  to C 10  alkenyl and alkynyl. Exemplary suitable modifications include O[(CH 2 ) n O] m CH 3 , O(CH 2 ). n OCH 3 , O(CH 2 ) n NH 2 , O(CH 2 ) n CH 3 , O(CH 2 ) n ONH 2 , and O(CH 2 ) n ON[(CH 2 ) n CH 3 )] 2 , where n and m are from 1 to about 10. In other embodiments, dsRNAs include one of the following at the 2′ position: C 1  to C 10  lower alkyl, substituted lower alkyl, alkaryl, aralkyl, O-alkaryl or O-aralkyl, SH, SCH 3 , OCN, Cl, Br, CN, CF 3 , OCF 3 , SOCH 3 , SO 2 CH 3 , ONO 2 , N 3 , NH 2 , heterocycloalkyl, heterocycloalkaryl, aminoalkylamino, polyalkylamino, substituted silyl, an RNA cleaving group, a reporter group, an intercalator, a group for improving the pharmacokinetic properties of a RNAi agent, or a group for improving the pharmacodynamic properties of a RNAi agent, and other substituents having similar properties. In some embodiments, the modification includes a 2′-methoxyethoxy (2′-O—CH 2 CH 2 OCH 3 , also known as 2′-O-(2-methoxyethyl) or 2′-MOE) (Martin et al.,  Helv. Chim. Acta,  1995, 78:486-504) i.e., an alkoxy-alkoxy group. Another exemplary modification is 2′-dimethylaminooxyethoxy, i.e., a O(CH 2 ) 2 ON(CH 3 ) 2  group, also known as 2′-DMAOE, as described in examples herein below, and 2′-dimethylaminoethoxyethoxy (also known in the art as 2′ dimethylaminoethoxyethyl or 2′-DMAEOE), i.e., 2′-O—CH 2 —O—CH 2 —N(CH 2 ) 2 . Further exemplary modifications include: 5′-Me-2′-F nucleotides, 5′-Me-2′-OMe nucleotides, 5′-Me-2′-deoxynucleotides, (both R and S isomers in these three families); 2′-alkoxyalkyl; and 2′-NMA (N-methylacetamide). 
     Other modifications include 2′-methoxy (2′-OCH 3 ), 2′-aminopropoxy (2′-OCH 2 CH 2 CH 2 NH 2 ), 2′-O-hexadecyl, and 2′-fluoro (2′-F). Similar modifications can also be made at other positions on the RNA of a RNAi agent, particularly the 3′ position of the sugar on the 3′ terminal nucleotide or in 2′-5′ linked dsRNAs and the 5′ position of 5′ terminal nucleotide. RNAi agents can also have sugar mimetics such as cyclobutyl moieties in place of the pentofuranosyl sugar. Representative U.S. patents that teach the preparation of such modified sugar structures include, but are not limited to, U.S. Pat. Nos. 4,981,957; 5,118,800; 5,319,080; 5,359,044; 5,393,878; 5,446,137; 5,466,786; 5,514,785; 5,519,134; 5,567,811; 5,576,427; 5,591,722; 5,597,909; 5,610,300; 5,627,053; 5,639,873; 5,646,265; 5,658,873; 5,670,633; and 5,700,920, certain of which are commonly owned with the instant application. The entire contents of each of the foregoing are hereby incorporated herein by reference. 
     An RNAi agent of the disclosure can also include nucleobase (often referred to in the art simply as “base”) modifications or substitutions. As used herein, “unmodified” or “natural” nucleobases include the purine bases adenine (A) and guanine (G), and the pyrimidine bases thymine (T), cytosine (C) and uracil (U). Modified nucleobases include other synthetic and natural nucleobases such as 5-methylcytosine (5-me-C), 5-hydroxymethyl cytosine, xanthine, hypoxanthine, 2-aminoadenine, 6-methyl and other alkyl derivatives of adenine and guanine, 2-propyl and other alkyl derivatives of adenine and guanine, 2-thiouracil, 2-thiothymine and 2-thiocytosine, 5-halouracil and cytosine, 5-propynyl uracil and cytosine, 6-azo uracil, cytosine and thymine, 5-uracil (pseudouracil), 4-thiouracil, 8-halo, 8-amino, 8-thiol, 8-thioalkyl, 8-hydroxyl anal other 8-substituted adenines and guanines, 5-halo, particularly 5-bromo, 5-trifluoromethyl and other 5-substituted uracils and cytosines, 7-methylguanine and 7-methyladenine, 8-azaguanine and 8-azaadenine, 7-deazaguanine and 7-daazaadenine and 3-deazaguanine and 3-deazaadenine. Further nucleobases include those disclosed in U.S. Pat. No. 3,687,808, those disclosed in Modified Nucleosides in Biochemistry, Biotechnology and Medicine, Herdewijn, P. ed. Wiley-VCH, 2008; those disclosed in The Concise Encyclopedia Of Polymer Science And Engineering, pages 858-859, Kroschwitz, J. L, ed. John Wiley &amp; Sons, 1990, these disclosed by Englisch et al., (1991)  Angewandte Chemie, International Edition,  30:613, and those disclosed by Sanghvi, Y S., Chapter 15, dsRNA Research and Applications, pages 289-302, Crooke, S. T. and Lebleu, B., Ed., CRC Press, 1993. Certain of these nucleobases are particularly useful for increasing the binding affinity of the oligomeric compounds featured in the disclosure. These include 5-substituted pyrimidines, 6-azapyrimidines and N-2, N-6 and 0-6 substituted purines, including 2-aminopropyladenine, 5-propynyluracil and 5-propynylcytosine. 5-methylcytosine substitutions have been shown to increase nucleic acid duplex stability by 0.6-1.2° C. (Sanghvi, Y. S., Crooke, S. T. and Lebleu, B., Eds., dsRNA Research and Applications, CRC Press, Boca Raton, 1993, pp. 276-278) and are exemplary base substitutions, even more particularly when combined with 2′-O-methoxyethyl sugar modifications. 
     Representative U.S. patents that teach the preparation of certain of the above noted modified nucleobases as well as other modified nucleobases include, but are not limited to, the above noted U.S. Pat. Nos. 3,687,808, 4,845,205; 5,130,30; 5,134,066; 5,175,273; 5,367,066; 5,432,272; 5,457,187; 5,459,255; 5,484,908; 5,502,177; 5,525,711; 5,552,540; 5,587,469; 5,594,121, 5,596,091; 5,614,617; 5,681,941; 5,750,692; 6,015,886; 6,147,200; 6,166,197; 6,222,025; 6,235,887; 6,380,368; 6,528,640; 6,639,062; 6,617,438; 7,045,610; 7,427,672; and 7,495,088, the entire contents of each of which are hereby incorporated herein by reference. 
     An RNAi agent of the disclosure can also be modified to include one or more locked nucleic acids (LNA). A locked nucleic acid is a nucleotide having a modified ribose moiety in which the ribose moiety comprises an extra bridge connecting the 2′ and 4′ carbons. This structure effectively “locks” the ribose in the 3′-endo structural conformation. The addition of locked nucleic acids to siRNAs has been shown to increase siRNA stability in serum, and to reduce off-target effects (Elmen, J. et al., (2005)  Nucleic Acids Research  33(1):439-447; Mook, O R. et al., (2007)  Mol Canc Ther  6(3):833-843; Grunweller, A. et al., (2003)  Nucleic Acids Research  31(12):3185-3193). 
     An RNAi agent of the disclosure can also be modified to include one or more bicyclic sugar moities. A “bicyclic sugar” is a furanosyl ring modified by the bridging of two atoms. A “bicyclic nucleoside” (“BNA”) is a nucleoside having a sugar moiety comprising a bridge connecting two carbon atoms of the sugar ring, thereby forming a bicyclic ring system. In certain embodiments, the bridge connects the 4′-carbon and the 2′-carbon of the sugar ring. Thus, in some embodiments an agent of the disclosure may include one or more locked nucleic acids (LNA). A locked nucleic acid is a nucleotide having a modified ribose moiety in which the ribose moiety comprises an extra bridge connecting the 2′ and 4′ carbons. In other words, an LNA is a nucleotide comprising a bicyclic sugar moiety comprising a 4′-CH2-O-2′ bridge. This structure effectively “locks” the ribose in the 3′-endo structural conformation. The addition of locked nucleic acids to siRNAs has been shown to increase siRNA stability in serum, and to reduce off-target effects (Elmen, J. et al., (2005)  Nucleic Acids Research  33(1):439-447; Mook, O R. et al., (2007)  Mol Canc Ther  6(3):833-843; Grunweller, A. et al., (2003)  Nucleic Acids Research  31(12):3185-3193). Examples of bicyclic nucleosides for use in the polynucleotides of the disclosure include without limitation nucleosides comprising a bridge between the 4′ and the 2′ ribosyl ring atoms. In certain embodiments, the antisense polynucleotide agents of the disclosure include one or more bicyclic nucleosides comprising a 4′ to 2′ bridge. Examples of such 4′ to 2′ bridged bicyclic nucleosides, include but are not limited to 4′-(CH 2 )—O-2′ (LNA); 4′-(CH2)-S-2′; 4′-(CH2) 2 —O-2′ (ENA); 4′-CH(CH3)-O-2′ (also referred to as “constrained ethyl” or “cEt”) and 4′-CH(CH2OCH3)-O-2′ (and analogs thereof; see, e.g., U.S. Pat. No. 7,399,845); 4′-C(CH3)(CH3)-O-2′ (and analogs thereof; see e.g., U.S. Pat. No. 8,278,283); 4′-CH2-N(OCH3)-2′ (and analogs thereof; see e.g., U.S. Pat. No. 8,278,425); 4′-CH2-O—N(CH3)-2′ (see, e.g., U.S. Patent Publication No. 2004/0171570); 4′-CH2-N(R)—O-2′, wherein R is H, C1-C12 alkyl, or a protecting group (see, e.g., U.S. Pat. No. 7,427,672); 4′-CH2-C(H)(CH3)-2′ (see, e.g., Chattopadhyaya et al.,  J. Org. Chem.,  2009, 74, 118-134); and 4′-CH2-C(═CH2)-2′ (and analogs thereof; see, e.g., U.S. Pat. No. 8,278,426). The entire contents of each of the foregoing are hereby incorporated herein by reference. 
     Additional representative US patents and US patenttent Publications that teach the preparation of locked nucleic acid nucleotides include, but are not limited to, the following: U.S. Pat. Nos. 6,268,490; 6,525,191; 6,670,461; 6,770,748; 6,794,499; 6,998,484; 7,053,207; 7,034,133; 7,084,125; 7,399,845; 7,427,672; 7,569,686; 7,741,457; 8,022,193; 8,030,467; 8,278,425; 8,278,426; 8,278,283; US 2008/0039618; and US 2009/0012281, the entire contents of each of which are hereby incorporated herein by reference. 
     Any of the foregoing bicyclic nucleosides can be prepared having one or more stereochemical sugar configurations including for example α-L-ribofuranose and β-D-ribofuranose (see WO 99/14226). 
     An RNAi agent of the disclosure can also be modified to include one or more constrained ethyl nucleotides. As used herein, a “constrained ethyl nucleotide” or “cEt” is a locked nucleic acid comprising a bicyclic sugar moiety comprising a 4′-CH(CH3)-O-2′ bridge. In one embodiment, a constrained ethyl nucleotide is in the S conformation referred to herein as “S-cEt.” 
     An RNAi agent of the disclosure may also include one or more “conformationally restricted nucleotides” (“CRN”). CRN are nucleotide analogs with a linker connecting the C2′ and C4′ carbons of ribose or the —C3′ and —C5′ carbons of ribose. CRN lock the ribose ring into a stable conformation and increase the hybridization affinity to mRNA. The linker is of sufficient length to place the oxygen in an optimal position for stability and affinity resulting in less ribose ring puckering. 
     Representative publications that teach the preparation of certain of the above noted CRN include, but are not limited to, US 2013/0190383; and WO 2013/036868, the entire contents of each of which are hereby incorporated herein by reference. 
     In some embodiments, a RNAi agent of the disclosure comprises one or more monomers that are UNA (unlocked nucleic acid) nucleotides. UNA is unlocked acyclic nucleic acid, wherein any of the bonds of the sugar has been removed, forming an unlocked “sugar” residue. In one example, UNA also encompasses monomer with bonds between C1′-C4′ have been removed (i.e. the covalent carbon-oxygen-carbon bond between the C1′ and C4′ carbons). In another example, the C2′-C3′ bond (i.e. the covalent carbon-carbon bond between the C2′ and C3′ carbons) of the sugar has been removed (see  Nuc. Acids Symp. Series,  52, 133-134 (2008) and Fluiter et al.,  Mol. Biosyst.,  2009, 10, 1039 hereby incorporated by reference). 
     Representative U.S. publications that teach the preparation of UNA include, but are not limited to, U.S. Pat. No. 8,314,227; and US Patent Publication Nos. 2013/0096289; 2013/0011922; and 2011/0313020, the entire contents of each of which are hereby incorporated herein by reference. 
     Potentially stabilizing modifications to the ends of RNA molecules can include N-(acetylaminocaproyl)-4-hydroxyprolinol (Hyp-C6-NHAc), N-(caproyl-4-hydroxyprolinol (Hyp-C6), N-(acetyl-4-hydroxyprolinol (Hyp-NHAc), thymidine-2′-O-deoxythymidine (ether), N-(aminocaproyl)-4-hydroxyprolinol (Hyp-C6-amino), 2-docosanoyl-uridine-3″-phosphate, inverted base dT(idT) and others. Disclosure of this modification can be found in WO 2011/005861. 
     Other modifications of a RNAi agent of the disclosure include a 5′ phosphate or 5′ phosphate mimic, e.g., a 5′-terminal phosphate or phosphate mimic on the antisense strand of a RNAi agent. Suitable phosphate mimics are disclosed in, for example US 2012/0157511, the entire contents of which are incorporated herein by reference. 
     A. Modified RNAi Agents Comprising Motifs of the Disclosure 
     In certain aspects of the disclosure, the double-stranded RNAi agents of the disclosure include agents with chemical modifications as disclosed, for example, in WO 2013/075035, the entire contents of which are incorporated herein by reference. As shown herein and in WO 2013/075035, a superior result may be obtained by introducing one or more motifs of three identical modifications on three consecutive nucleotides into a sense strand or antisense strand of an RNAi agent, particularly at or near the cleavage site. In some embodiments, the sense strand and antisense strand of the RNAi agent may otherwise be completely modified. The introduction of these motifs interrupts the modification pattern, if present, of the sense or antisense strand. The RNAi agent may be optionally conjugated with a lipophilic ligand, e.g., a C16 ligand, for instance on the sense strand. The RNAi agent may be optionally modified with a (S)-glycol nucleic acid (GNA) modification, for instance on one or more residues of the antisense strand. The resulting RNAi agents present superior gene silencing activity. 
     Accordingly, the disclosure provides double stranded RNAi agents capable of inhibiting the expression of a target genome or gene (i.e., an ACE2 gene) in vivo. The RNAi agent comprises a sense strand and an antisense strand. Each strand of the RNAi agent may be 15-30 nucleotides in length. For example, each strand may be 16-30 nucleotides in length, 17-30 nucleotides in length, 25-30 nucleotides in length, 27-30 nucleotides in length, 17-23 nucleotides in length, 17-21 nucleotides in length, 17-19 nucleotides in length, 19-25 nucleotides in length, 19-23 nucleotides in length, 19-21 nucleotides in length, 21-25 nucleotides in length, or 21-23 nucleotides in length. In certain embodiments, each strand is 19-23 nucleotides in length. 
     The sense strand and antisense strand typically form a duplex double stranded RNA (“dsRNA”), also referred to herein as an “RNAi agent.” The duplex region of an RNAi agent may be 15-30 nucleotide pairs in length. For example, the duplex region can be 16-30 nucleotide pairs in length, 17-30 nucleotide pairs in length, 27-30 nucleotide pairs in length, 17-23 nucleotide pairs in length, 17-21 nucleotide pairs in length, 17-19 nucleotide pairs in length, 19-25 nucleotide pairs in length, 19-23 nucleotide pairs in length, 19-21 nucleotide pairs in length, 21-25 nucleotide pairs in length, or 21-23 nucleotide pairs in length. In another example, the duplex region is selected from 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, and 27 nucleotides in length. In preferred embodiments, the duplex region is 19-21 nucleotide pairs in length. 
     In one embodiment, the RNAi agent may contain one or more overhang regions or capping groups at the 3′-end, 5′-end, or both ends of one or both strands. The overhang can be 1-6 nucleotides in length, for instance 2-6 nucleotides in length, 1-5 nucleotides in length, 2-5 nucleotides in length, 1-4 nucleotides in length, 2-4 nucleotides in length, 1-3 nucleotides in length, 2-3 nucleotides in length, or 1-2 nucleotides in length. In preferred embodiments, the nucleotide overhang region is 2 nucleotides in length. The overhangs can be the result of one strand being longer than the other, or the result of two strands of the same length being staggered. The overhang can form a mismatch with the target mRNA or it can be complementary to the gene sequences being targeted or can be another sequence. The first and second strands can also be joined, e.g., by additional bases to form a hairpin, or by other non-base linkers. 
     In one embodiment, the nucleotides in the overhang region of the RNAi agent can each independently be a modified or unmodified nucleotide including, but no limited to 2′-sugar modified, such as, 2-F, 2′-O-methyl, thymidine (T), and any combinations thereof. 
     For example, TT can be an overhang sequence for either end on either strand. The overhang can form a mismatch with the target mRNA or it can be complementary to the gene sequences being targeted or can be another sequence. 
     The 5′- or 3′-overhangs at the sense strand, antisense strand or both strands of the RNAi agent may be phosphorylated. In some embodiments, the overhang region(s) contains two nucleotides having a phosphorothioate between the two nucleotides, where the two nucleotides can be the same or different. In one embodiment, the overhang is present at the 3′-end of the sense strand, antisense strand, or both strands. In one embodiment, this 3′-overhang is present in the antisense strand. In one embodiment, this 3′-overhang is present in the sense strand. 
     The RNAi agent may contain only a single overhang, which can strengthen the interference activity of the RNAi, without affecting its overall stability. For example, the single-stranded overhang may be located at the 3′-terminal end of the sense strand or, alternatively, at the 3′-terminal end of the antisense strand. The RNAi may also have a blunt end, located at the 5′-end of the antisense strand (or the 3′-end of the sense strand) or vice versa. Generally, the antisense strand of the RNAi has a nucleotide overhang at the 3′-end, and the 5′-end is blunt. While not wishing to be bound by theory, the asymmetric blunt end at the 5′-end of the antisense strand and 3′-end overhang of the antisense strand favor the guide strand loading into RISC process. 
     In one embodiment, the RNAi agent is a double ended bluntmer of 19 nucleotides in length, wherein the sense strand contains at least one motif of three 2′-F modifications on three consecutive nucleotides at positions 7, 8, 9 from the 5′end. The antisense strand contains at least one motif of three 2′-O-methyl modifications on three consecutive nucleotides at positions 11, 12, 13 from the 5′end. 
     In another embodiment, the RNAi agent is a double ended bluntmer of 20 nucleotides in length, wherein the sense strand contains at least one motif of three 2′-F modifications on three consecutive nucleotides at positions 8, 9, 10 from the 5′end. The antisense strand contains at least one motif of three 2′-O-methyl modifications on three consecutive nucleotides at positions 11, 12, 13 from the 5′end. 
     In yet another embodiment, the RNAi agent is a double ended bluntmer of 21 nucleotides in length, wherein the sense strand contains at least one motif of three 2′-F modifications on three consecutive nucleotides at positions 9, 10, 11 from the 5′end. The antisense strand contains at least one motif of three 2′-O-methyl modifications on three consecutive nucleotides at positions 11, 12, 13 from the 5′end. 
     In one embodiment, the RNAi agent comprises a 21 nucleotide sense strand and a 23 nucleotide antisense strand, wherein the sense strand contains at least one motif of three 2′-F modifications on three consecutive nucleotides at positions 9, 10, 11 from the 5′ end; the antisense strand contains at least one motif of three 2′-O-methyl modifications on three consecutive nucleotides at positions 11, 12, 13 from the 5′end, wherein one end of the RNAi agent is blunt, while the other end comprises a 2 nucleotide overhang. Preferably, the 2 nucleotide overhang is at the 3′-end of the antisense strand. When the 2 nucleotide overhang is at the 3′-end of the antisense strand, there may be two phosphorothioate internucleotide linkages between the terminal three nucleotides, wherein two of the three nucleotides are the overhang nucleotides, and the third nucleotide is a paired nucleotide next to the overhang nucleotide. In one embodiment, the RNAi agent additionally has two phosphorothioate internucleotide linkages between the terminal three nucleotides at both the 5′-end of the sense strand and at the 5′-end of the antisense strand. In one embodiment, every nucleotide in the sense strand and the antisense strand of the RNAi agent, including the nucleotides that are part of the motifs are modified nucleotides. In one embodiment each residue is independently modified with a 2′-O-methyl or 3′-fluoro, e.g., in an alternating motif. Optionally, the RNAi agent further comprises a ligand (e.g., a lipophilic ligand, optionally a C16 ligand). 
     In one embodiment, the RNAi agent comprises a sense and an antisense strand, wherein the sense strand is 25-30 nucleotide residues in length, wherein starting from the 5′ terminal nucleotide (position 1) positions 1 to 23 of the first strand comprise at least 8 ribonucleotides; the antisense strand is 36-66 nucleotide residues in length and, starting from the 3′ terminal nucleotide, comprises at least 8 ribonucleotides in the positions paired with positions 1-23 of sense strand to form a duplex; wherein at least the 3 ‘ terminal nucleotide of antisense strand is unpaired with sense strand, and up to 6 consecutive 3’ terminal nucleotides are unpaired with sense strand, thereby forming a 3′ single stranded overhang of 1-6 nucleotides; wherein the 5′ terminus of antisense strand comprises from 10-30 consecutive nucleotides which are unpaired with sense strand, thereby forming a 10-30 nucleotide single stranded 5′ overhang; wherein at least the sense strand 5′ terminal and 3′ terminal nucleotides are base paired with nucleotides of antisense strand when sense and antisense strands are aligned for maximum complementarity, thereby forming a substantially duplexed region between sense and antisense strands; and antisense strand is sufficiently complementary to a target RNA along at least 19 ribonucleotides of antisense strand length to reduce target gene expression when the double stranded nucleic acid is introduced into a mammalian cell; and wherein the sense strand contains at least one motif of three 2′-F modifications on three consecutive nucleotides, where at least one of the motifs occurs at or near the cleavage site. The antisense strand contains at least one motif of three 2′ methyl modifications on three consecutive nucleotides at or near the cleavage site. 
     In one embodiment, the RNAi agent comprises sense and antisense strands, wherein the RNAi agent comprises a first strand having a length which is at least 25 and at most 29 nucleotides and a second strand having a length which is at most 30 nucleotides with at least one motif of three 2′-O-methyl modifications on three consecutive nucleotides at position 11, 12, 13 from the 5′ end; wherein the 3′ end of the first strand and the 5′ end of the second strand form a blunt end and the second strand is 1-4 nucleotides longer at its 3′ end than the first strand, wherein the duplex region region which is at least 25 nucleotides in length, and the second strand is sufficiently complementary to a target mRNA along at least 19 nucleotide of the second strand length to reduce target gene expression when the RNAi agent is introduced into a mammalian cell, and wherein dicer cleavage of the RNAi agent preferentially results in an siRNA comprising the 3′ end of the second strand, thereby reducing expression of the target gene in the mammal. Optionally, the RNAi agent further comprises a ligand. 
     In one embodiment, the sense strand of the RNAi agent contains at least one motif of three identical modifications on three consecutive nucleotides, where one of the motifs occurs at the cleavage site in the sense strand. 
     In one embodiment, the antisense strand of the RNAi agent can also contain at least one motif of three identical modifications on three consecutive nucleotides, where one of the motifs occurs at or near the cleavage site in the antisense strand. 
     For an RNAi agent having a duplex region of 17-23 nucleotide in length, the cleavage site of the antisense strand is typically around the 10, 11 and 12 positions from the 5′-end. Thus the motifs of three identical modifications may occur at the 9, 10, 11 positions; 10, 11, 12 positions; 11, 12, 13 positions; 12, 13, 14 positions; or 13, 14, 15 positions of the antisense strand, the count starting from the 1 st  nucleotide from the 5′-end of the antisense strand, or, the count starting from the 1 st  paired nucleotide within the duplex region from the 5′-end of the antisense strand. The cleavage site in the antisense strand may also change according to the length of the duplex region of the RNAi from the 5′-end. 
     The sense strand of the RNAi agent may contain at least one motif of three identical modifications on three consecutive nucleotides at the cleavage site of the strand; and the antisense strand may have at least one motif of three identical modifications on three consecutive nucleotides at or near the cleavage site of the strand. When the sense strand and the antisense strand form a dsRNA duplex, the sense strand and the antisense strand can be so aligned that one motif of the three nucleotides on the sense strand and one motif of the three nucleotides on the antisense strand have at least one nucleotide overlap, i.e., at least one of the three nucleotides of the motif in the sense strand forms a base pair with at least one of the three nucleotides of the motif in the antisense strand. Alternatively, at least two nucleotides may overlap, or all three nucleotides may overlap. 
     In one embodiment, the sense strand of the RNAi agent may contain more than one motif of three identical modifications on three consecutive nucleotides. The first motif may occur at or near the cleavage site of the strand and the other motifs may be a wing modification. The term “wing modification” herein refers to a motif occurring at another portion of the strand that is separated from the motif at or near the cleavage site of the same strand. The wing modification is either adjacent to the first motif or is separated by at least one or more nucleotides. When the motifs are immediately adjacent to each other then the chemistry of the motifs are distinct from each other and when the motifs are separated by one or more nucleotide than the chemistries can be the same or different. Two or more wing modifications may be present. For instance, when two wing modifications are present, each wing modification may occur at one end relative to the first motif which is at or near cleavage site or on either side of the lead motif. 
     Like the sense strand, the antisense strand of the RNAi agent may contain more than one motif of three identical modifications on three consecutive nucleotides, with at least one of the motifs occurring at or near the cleavage site of the strand. This antisense strand may also contain one or more wing modifications in an alignment similar to the wing modifications that may be present on the sense strand. 
     In one embodiment, the wing modification on the sense strand or antisense strand of the RNAi agent typically does not include the first one or two terminal nucleotides at the 3′-end, 5′-end or both ends of the strand. 
     In another embodiment, the wing modification on the sense strand or antisense strand of the RNAi agent typically does not include the first one or two paired nucleotides within the duplex region at the 3′-end, 5′-end or both ends of the strand. 
     When the sense strand and the antisense strand of the RNAi agent each contain at least one wing modification, the wing modifications may fall on the same end of the duplex region, and have an overlap of one, two or three nucleotides. 
     When the sense strand and the antisense strand of the RNAi agent each contain at least two wing modifications, the sense strand and the antisense strand can be so aligned that two modifications each from one strand fall on one end of the duplex region, having an overlap of one, two or three nucleotides; two modifications each from one strand fall on the other end of the duplex region, having an overlap of one, two or three nucleotides; two modifications one strand fall on each side of the lead motif, having an overlap of one, two, or three nucleotides in the duplex region. 
     In one embodiment, the RNAi agent comprises mismatch(es) with the target, within the duplex, or combinations thereof. The mistmatch may occur in the overhang region or the duplex region. The base pair may be ranked on the basis of their propensity to promote dissociation or melting (e.g., on the free energy of association or dissociation of a particular pairing, the simplest approach is to examine the pairs on an individual pair basis, though next neighbor or similar analysis can also be used). In terms of promoting dissociation: A:U is preferred over G:C; G:U is preferred over G:C; and I:C is preferred over G:C (I=inosine). Mismatches, e.g., non-canonical or other than canonical pairings (as described elsewhere herein) are preferred over canonical (A:T, A:U, G:C) pairings; and pairings which include a universal base are preferred over canonical pairings. 
     In one embodiment, the RNAi agent comprises at least one of the first 1, 2, 3, 4, or 5 base pairs within the duplex regions from the 5′-end of the antisense strand independently selected from the group of: A:U, G:U, I:C, and mismatched pairs, e.g., non-canonical or other than canonical pairings or pairings which include a universal base, to promote the dissociation of the antisense strand at the 5′-end of the duplex. 
     In one embodiment, the nucleotide at the 1 position within the duplex region from the 5′-end in the antisense strand is selected from the group consisting of A, dA, dU, U, and dT. Alternatively, at least one of the first 1, 2 or 3 base pair within the duplex region from the 5′-end of the antisense strand is an AU base pair. For example, the first base pair within the duplex region from the 5′-end of the antisense strand is an AU base pair. 
     In another embodiment, the nucleotide at the 3′-end of the sense strand is deoxy-thymine (dT). In another embodiment, the nucleotide at the 3′-end of the antisense strand is deoxy-thymine (dT). In one embodiment, there is a short sequence of deoxy-thymine nucleotides, for example, two dT nucleotides on the 3′-end of the sense or antisense strand. 
     In one embodiment, the sense strand sequence may be represented by formula (I): 
       5′n p -N a -(XXX) i -N b -YYY-N b -(ZZZ) j -N a -n q 3′  (I)
 
     wherein: 
     i and j are each independently 0 or 1; 
     p and q are each independently 0-6; 
     each N a  independently represents an oligonucleotide sequence comprising 0-25 modified nucleotides, each sequence comprising at least two differently modified nucleotides; 
     each N b  independently represents an oligonucleotide sequence comprising 0-10 modified nucleotides; 
     each n p  and n q  independently represent an overhang nucleotide; 
     wherein N b  and Y do not have the same modification; and 
     XXX, YYY and ZZZ each independently represent one motif of three identical modifications on three consecutive nucleotides. Preferably YYY is all 2′-F modified nucleotides. 
     In one embodiment, the N a  or N b  comprise modifications of alternating pattern. 
     In one embodiment, the YYY motif occurs at or near the cleavage site of the sense strand. For example, when the RNAi agent has a duplex region of 17-23 nucleotides in length, the YYY motif can occur at or the vicinity of the cleavage site (e.g.: can occur at positions 6, 7, 8, 7, 8, 9, 8, 9, 10, 9, 10, 11, 10, 11,12 or 11, 12, 13) of-the sense strand, the count starting from the 1 st  nucleotide, from the 5′-end; or optionally, the count starting at the 1 st  paired nucleotide within the duplex region, from the 5′-end. 
     In one embodiment, i is 1 and j is 0, or i is 0 and j is 1, or both i and j are 1. The sense strand can therefore be represented by the following formulas: 
       5′n p -N a -YYY-N b -ZZZ-N a -n q 3′  (Ib);
 
       5′n p -N a -XXX-N b -YYY-N a -n q 3′  (Ic); or
 
       5′n p -N a -XXX-N b -YYY-N b -ZZZ-N a -n q 3′  (Id).
 
     When the sense strand is represented by formula (Ib), N b  represents an oligonucleotide sequence comprising 0-10, 0-7, 0-5, 0-4, 0-2 or 0 modified nucleotides. 
     Each N a  independently can represent an oligonucleotide sequence comprising 2-20, 2-15, or 2-10 modified nucleotides. 
     When the sense strand is represented as formula (Ic), N b  represents an oligonucleotide sequence comprising 0-10, 0-7, 0-10, 0-7, 0-5, 0-4, 0-2 or 0 modified nucleotides. Each N a  can independently represent an oligonucleotide sequence comprising 2-20, 2-15, or 2-10 modified nucleotides. 
     When the sense strand is represented as formula (Id), each N b  independently represents an oligonucleotide sequence comprising 0-10, 0-7, 0-5, 0-4, 0-2 or 0 modified nucleotides. Preferably, N b  is 0, 1, 2, 3, 4, 5 or 6. Each N a  can independently represent an oligonucleotide sequence comprising 2-20, 2-15, or 2-10 modified nucleotides. 
     Each of X, Y and Z may be the same or different from each other. 
     In other embodiments, i is 0 and j is 0, and the sense strand may be represented by the formula: 
       5′n p -N a -YYY-N a -n q 3′  (Ia).
 
     When the sense strand is represented by formula (Ia), each N a  independently can represent an oligonucleotide sequence comprising 2-20, 2-15, or 2-10 modified nucleotides. 
     In one embodiment, the antisense strand sequence of the RNAi may be represented by formula (II): 
       5′n q′ -N a ′-(Z′Z′Z′) k -N b ′-Y′Y′Y′-N b ′-(X′X′X′) l -N′ a -n p ′3′  (II)
 
     wherein: 
     k and l are each independently 0 or 1; 
     p′ and q′ are each independently 0-6; 
     each N a ′ independently represents an oligonucleotide sequence comprising 0-25 modified nucleotides, each sequence comprising at least two differently modified nucleotides;
 
each N b ′ independently represents an oligonucleotide sequence comprising 0-10 modified nucleotides;
 
each n p ′ and n q ′ independently represent an overhang nucleotide;
 
wherein N b ′ and Y′ do not have the same modification; and
 
X′X′X′, Y′Y′Y′ and Z′Z′Z′ each independently represent one motif of three identical modifications on three consecutive nucleotides.
 
     In one embodiment, the N a ′ or N b ′ comprise modifications of alternating pattern. 
     The Y′Y′Y′ motif occurs at or near the cleavage site of the antisense strand. For example, when the RNAi agent has a duplex region of 17-23 nucleotide in length, the Y′Y′Y′ motif can occur at positions 9, 10, 11; 10, 11, 12; 11, 12, 13; 12, 13, 14; or 13, 14, 15 of the antisense strand, with the count starting from the 1 st  nucleotide, from the 5′-end; or optionally, the count starting at the 1 st  paired nucleotide within the duplex region, from the 5′-end. Preferably, the Y′Y′Y′ motif occurs at positions 11, 12, 13. 
     In one embodiment, Y′Y′Y′ motif is all 2′-OMe modified nucleotides. 
     In one embodiment, k is 1 and l is 0, or k is 0 and l is 1, or both k and l are 1. 
     The antisense strand can therefore be represented by the following formulas: 
       5′n q′ -N a ′-Z′Z′Z′-N b ′-Y′Y′Y′-N a ′-n p′ 3′  (IIb);
 
       5′n q′ -N a ′-Y′Y′Y′-N b ′-X′X′X′-n p′ 3′  (IIc); or
 
       5′n q′ -N b ′-Z′Z′Z′-N b ′-Y′Y′Y′-N b ′-X′X′X′-N a ′-n p′ 3′  (IId).
 
     When the antisense strand is represented by formula (IIb), N b ′ represents an oligonucleotide sequence comprising 0-10, 0-7, 0-10, 0-7, 0-5, 0-4, 0-2 or 0 modified nucleotides. Each N a ′ independently represents an oligonucleotide sequence comprising 2-20, 2-15, or 2-10 modified nucleotides. 
     When the antisense strand is represented as formula (IIc), N b ′ represents an oligonucleotide sequence comprising 0-10, 0-7, 0-10, 0-7, 0-5, 0-4, 0-2 or 0 modified nucleotides. Each N a ′ independently represents an oligonucleotide sequence comprising 2-20, 2-15, or 2-10 modified nucleotides. 
     When the antisense strand is represented as formula (IId), each N b ′ independently represents an oligonucleotide sequence comprising 0-10, 0-7, 0-10, 0-7, 0-5, 0-4, 0-2 or 0 modified nucleotides. Each N a ′ independently represents an oligonucleotide sequence comprising 2-20, 2-15, or 2-10 modified nucleotides. Preferably, N b  is 0, 1, 2, 3, 4, 5 or 6. 
     In other embodiments, k is 0 and l is 0 and the antisense strand may be represented by the formula: 
       5′n p′ -N a′ -Y′Y′Y′-N a′ -n q′ 3′  (Ia).
 
     When the antisense strand is represented as formula (Ha), each N a ′ independently represents an oligonucleotide sequence comprising 2-20, 2-15, or 2-10 modified nucleotides. 
     Each of X′, Y′ and Z′ may be the same or different from each other. 
     Each nucleotide of the sense strand and antisense strand may be independently modified with LNA, HNA, CeNA, 2′-methoxyethyl, 2′-O-methyl, 2′-O-allyl, 2′-C-allyl, 2′-hydroxyl, or 2′-fluoro. For example, each nucleotide of the sense strand and antisense strand is independently modified with 2′-O-methyl or 2′-fluoro. Each X, Y, Z, X′, Y′ and Z′, in particular, may represent a 2′-O-methyl modification or a 2′-fluoro modification. 
     In one embodiment, the sense strand of the RNAi agent may contain YYY motif occurring at 9, 10 and 11 positions of the strand when the duplex region is 21 nt, the count starting from the 1st nucleotide from the 5′-end, or optionally, the count starting at the 1 st  paired nucleotide within the duplex region, from the 5′-end; and Y represents 2′-F modification. The sense strand may additionally contain XXX motif or ZZZ motifs as wing modifications at the opposite end of the duplex region; and XXX and ZZZ each independently represents a 2′-OMe modification or 2′-F modification. 
     In one embodiment the antisense strand may contain Y′Y′Y′ motif occurring at positions 11, 12, 13 of the strand, the count starting from the 1 st  nucleotide from the 5′-end, or optionally, the count starting at the 1 st  paired nucleotide within the duplex region, from the 5′-end; and Y′ represents 2′-O-methyl modification. The antisense strand may additionally contain X′X′X′ motif or Z′Z′Z′ motifs as wing modifications at the opposite end of the duplex region; and X′X′X′ and Z′Z′Z′ each independently represents a 2′-OMe modification or 2′-F modification. 
     The sense strand represented by any one of the above formulas (Ia), (Ib), (Ic), and (Id) forms a duplex with a antisense strand being represented by any one of formulas (IIa), (IIb), (IIc), and (IId), respectively. 
     Accordingly, the RNAi agents for use in the methods of the disclosure may comprise a sense strand and an antisense strand, each strand having 14 to 30 nucleotides, the RNAi duplex represented by formula (III): 
       sense:5′n p -N a -(XXX) i -N b -YYY-N b -(ZZZ) j -N a -n q 3′
 
       antisense:3′n p -N a -(X′X′X′) k -N b ′-Y′Y′Y′-N b ′-(Z′Z′Z′) l -N a ′-n q ′5′  (III)
 
     wherein: 
     j, k, and l are each independently 0 or 1; 
     p, p′, q, and q′ are each independently 0-6; 
     each N a  and N a ′ independently represents an oligonucleotide sequence comprising 0-25 modified nucleotides, each sequence comprising at least two differently modified nucleotides; 
     each N b  and N b ′ independently represents an oligonucleotide sequence comprising 0-10 modified nucleotides; 
     wherein 
     each n p ′, n p , n q ′, and n q , each of which may or may not be present, independently represents an overhang nucleotide; and 
     XXX, YYY, ZZZ, X′X′X′, Y′Y′Y′, and Z′Z′Z′ each independently represent one motif of three identical modifications on three consecutive nucleotides. 
     In one embodiment, i is 0 and j is 0; or i is 1 and j is 0; or i is 0 and j is 1; or both i and j are 0; or both i and j are 1. In another embodiment, k is 0 and l is 0; or k is 1 and l is 0; k is 0 and l is 1; or both k and l are 0; or both k and l are 1. 
     Exemplary combinations of the sense strand and antisense strand forming a RNAi duplex include the formulas below: 
       5′n p -N a -YYY-N a -n q 3′
 
       3′n p ′-N a ′-Y′Y′Y′-N a ′n q ′5′  (IIIa)
 
       5′n p -N a -YYY-N b -ZZZ-N a -n q 3′
 
       3′n p ′-N a ′-Y′Y′Y′-N b ′-Z′Z′Z′-N a ′n q ′5′  (IIIb)
 
       5′n p -N a -XXX-N b -YYY-N a -n q 3′
 
       3′n p ′-N a ′-X′X′X′-N b ′-Y′Y′Y′-N a ′-n q ′5′  (IIIc)
 
       5′n p -N a -XXX-N b -YYY-N b -XXX-N a -n q 3′
 
       3′n p ′-N a ′-X′X′X′-N b ′-Y′Y′Y′-N b ′-Z′Z′Z′-N a -n q ′5′  (IIId)
 
     When the RNAi agent is represented by formula (IIIa), each N a  independently represents an oligonucleotide sequence comprising 2-20, 2-15, or 2-10 modified nucleotides. 
     When the RNAi agent is represented by formula (IIIb), each N b  independently represents an oligonucleotide sequence comprising 1-10, 1-7, 1-5 or 1-4 modified nucleotides. Each N a  independently represents an oligonucleotide sequence comprising 2-20, 2-15, or 2-10 modified nucleotides. 
     When the RNAi agent is represented as formula (IIIc), each N b , N b ′ independently represents an oligonucleotide sequence comprising 0-10, 0-7, 0-10, 0-7, 0-5, 0-4, 0-2 or 0 modified nucleotides. Each N a  independently represents an oligonucleotide sequence comprising 2-20, 2-15, or 2-10 modified nucleotides. 
     When the RNAi agent is represented as formula (IIId), each N b , N b ′ independently represents an oligonucleotide sequence comprising 0-10, 0-7, 0-10, 0-7, 0-5, 0-4, 0-2 or 0 modified nucleotides. 
     Each N a , N a ′ independently represents an oligonucleotide sequence comprising 2-20, 2-15, or 2-10 modified nucleotides. Each of N a , N a ′, N b  and N b ′ independently comprises modifications of alternating pattern. 
     In one embodiment, when the RNAi agent is represented by formula (IIId), the N a  modifications are 2′-O-methyl or 2′-fluoro modifications. In another embodiment, when the RNAi agent is represented by formula (IIId), the N a  modifications are 2′-O-methyl or 2′-fluoro modifications and n p ′&gt;0 and at least one n p ′ is linked to a neighboring nucleotide a via phosphorothioate linkage. In yet another embodiment, when the RNAi agent is represented by formula (IIId), the N a  modifications are 2′-O-methyl or 2′-fluoro modifications, n p ′&gt;0 and at least one n p ′ is linked to a neighboring nucleotide via phosphorothioate linkage, and the sense strand is conjugated to one or more C16 (or related) moieties attached through a bivalent or trivalent branched linker (described below). In another embodiment, when the RNAi agent is represented by formula (IIId), the N a  modifications are 2′-O-methyl or 2′-fluoro modifications, n p ′&gt;0 and at least one n p ′ is linked to a neighboring nucleotide via phosphorothioate linkage, the sense strand comprises at least one phosphorothioate linkage, and the sense strand is conjugated to one or more lipophilic, e.g., C16 (or related) moieties, optionally attached through a bivalent or trivalent branched linker. 
     In one embodiment, when the RNAi agent is represented by formula (IIIa), the N a  modifications are 2′-O-methyl or 2′-fluoro modifications, n p ′&gt;0 and at least one n p ′ is linked to a neighboring nucleotide via phosphorothioate linkage, the sense strand comprises at least one phosphorothioate linkage, and the sense strand is conjugated to one or more lipophilic, e.g., C16 (or related) moieties attached through a bivalent or trivalent branched linker. 
     In one embodiment, the RNAi agent is a multimer containing at least two duplexes represented by formula (III), (IIIa), (IIIb), (IIIc), and (IIId), wherein the duplexes are connected by a linker. The linker can be cleavable or non-cleavable. Optionally, the multimer further comprises a ligand. Each of the duplexes can target the same gene or two different genes; or each of the duplexes can target same gene at two different target sites. 
     In one embodiment, the RNAi agent is a multimer containing three, four, five, six or more duplexes represented by formula (III), (IIIa), (IIIb), (IIIc), and (IIId), wherein the duplexes are connected by a linker. The linker can be cleavable or non-cleavable. Optionally, the multimer further comprises a ligand. Each of the duplexes can target the same gene or two different genes; or each of the duplexes can target same gene at two different target sites. 
     In one embodiment, two RNAi agents represented by formula (III), (IIIa), (Mb), (IIIc), and (IIId) are linked to each other at the 5′ end, and one or both of the 3′ ends and are optionally conjugated to to a ligand. Each of the agents can target the same gene or two different genes; or each of the agents can target same gene at two different target sites. 
     Various publications describe multimeric RNAi agents that can be used in the methods of the disclosure. Such publications include WO2007/091269, WO2010/141511, WO2007/117686, WO2009/014887, and WO2011/031520; and U.S. Pat. No. 7,858,769, the entire contents of each of which are hereby incorporated herein by reference. 
     In certain embodiments, the compositions and methods of the disclosure include a vinyl phosphonate (VP) modification of an RNAi agent as described herein. In exemplary embodiments, a vinyl phosphonate of the disclosure has the following structure: 
     
       
         
         
             
             
         
       
     
     A vinyl phosphonate of the instant disclosure may be attached to either the antisense or the sense strand of a dsRNA of the disclosure. In certain preferred embodiments, a vinyl phosphonate of the instant disclosure is attached to the antisense strand of a dsRNA, optionally at the 5′ end of the antisense strand of the dsRNA. 
     Vinyl phosphate modifications are also contemplated for the compositions and methods of the instant disclosure. An exemplary vinyl phosphate structure is: 
     
       
         
         
             
             
         
       
     
     E. Thermally Destabilizing Modifications 
     In certain embodiments, a dsRNA molecule can be optimized for RNA interference by incorporating thermally destabilizing modifications in the seed region of the antisense strand (i.e., at positions 2-9 of the 5′-end of the antisense strand) to reduce or inhibit off-target gene silencing. It has been discovered that dsRNAs with an antisense strand comprising at least one thermally destabilizing modification of the duplex within the first 9 nucleotide positions, counting from the 5′ end, of the antisense strand have reduced off-target gene silencing activity. Accordingly, in some embodiments, the antisense strand comprises at least one (e.g., one, two, three, four, five or more) thermally destabilizing modification of the duplex within the first 9 nucleotide positions of the 5′ region of the antisense strand. In some embodiments, one or more thermally destabilizing modification(s) of the duplex is/are located in positions 2-9, or preferably positions 4-8, from the 5′-end of the antisense strand. In some further embodiments, the thermally destabilizing modification(s) of the duplex is/are located at position 6, 7 or 8 from the 5′-end of the antisense strand. In still some further embodiments, the thermally destabilizing modification of the duplex is located at position 7 from the 5′-end of the antisense strand. The term “thermally destabilizing modification(s)” includes modification(s) that would result with a dsRNA with a lower overall melting temperature (Tm) (preferably a Tm with one, two, three or four degrees lower than the Tm of the dsRNA without having such modification(s). In some embodiments, the thermally destabilizing modification of the duplex is located at position 2, 3, 4, 5 or 9 from the 5′-end of the antisense strand. 
     The thermally destabilizing modifications can include, but are not limited to, abasic modification; mismatch with the opposing nucleotide in the opposing strand; and sugar modification such as 2′-deoxy modification or acyclic nucleotide, e.g., unlocked nucleic acids (UNA) or glycol nucleic acid (GNA). 
     Exemplified abasic modifications include, but are not limited to the following: 
     
       
         
         
             
             
         
       
     
     Wherein R=H, Me, Et or OMe; R′=H, Me, Et or OMe; R″=H, Me, Et or OMe 
     
       
         
         
             
             
         
       
     
     wherein B is a modified or unmodified nucleobase. 
     Exemplified sugar modifications include, but are not limited to the following: 
     
       
         
         
             
             
         
       
     
     wherein B is a modified or unmodified nucleobase. 
     In some embodiments the thermally destabilizing modification of the duplex is selected from the group consisting of: 
     
       
         
         
             
             
         
       
     
     wherein B is a modified or unmodified nucleobase and the asterisk on each structure represents either R, S or racemic. 
     The term “acyclic nucleotide” refers to any nucleotide having an acyclic ribose sugar, for example, where any of bonds between the ribose carbons (e.g., C1′-C2′, C2′-C3′, C3′-C4′, C4′-C4′, or C1′-C4′) is absent or at least one of ribose carbons or oxygen (e.g., C1′, C2′, C3′, C4′, or O4′) are independently or in combination absent from the nucleotide. In some embodiments, acyclic nucleotide is 
     
       
         
         
             
             
         
       
     
     wherein B is a modified or unmodified nucleobase, R 1  and R 2  independently are H, halogen, OR 3 , or alkyl; and R 3  is H, alkyl, cycloalkyl, aryl, aralkyl, heteroaryl or sugar). The term “UNA” refers to unlocked acyclic nucleic acid, wherein any of the bonds of the sugar has been removed, forming an unlocked “sugar” residue. In one example, UNA also encompasses monomers with bonds between C1′-C4′ being removed (i.e. the covalent carbon-oxygen-carbon bond between the C1′ and C4′ carbons). In another example, the C2′-C3′ bond (i.e. the covalent carbon-carbon bond between the C2′ and C3′ carbons) of the sugar is removed (see Mikhailov et. al., Tetrahedron Letters, 26 (17): 2059 (1985); and Fluiter et al., Mol. Biosyst., 10: 1039 (2009), which are hereby incorporated by reference in their entirety). The acyclic derivative provides greater backbone flexibility without affecting the Watson-Crick pairings. The acyclic nucleotide can be linked via 2′-5′ or 3′-5′ linkage. 
     The term ‘GNA’ refers to glycol nucleic acid which is a polymer similar to DNA or RNA but differing in the composition of its “backbone” in that is composed of repeating glycerol units linked by phosphodiester bonds: 
     
       
         
         
             
             
         
       
     
     The thermally destabilizing modification of the duplex can be mismatches (i.e., noncomplementary base pairs) between the thermally destabilizing nucleotide and the opposing nucleotide in the opposite strand within the dsRNA duplex. Exemplary mismatch base pairs include G:G, G:A, G:U, G:T, A:A, A:C, C:C, C:U, C:T, U:U, T:T, U:T, or a combination thereof. Other mismatch base pairings known in the art are also amenable to the present invention. A mismatch can occur between nucleotides that are either naturally occurring nucleotides or modified nucleotides, i.e., the mismatch base pairing can occur between the nucleobases from respective nucleotides independent of the modifications on the ribose sugars of the nucleotides. In certain embodiments, the dsRNA molecule contains at least one nucleobase in the mismatch pairing that is a 2′-deoxy nucleobase; e.g., the 2′-deoxy nucleobase is in the sense strand. 
     In some embodiments, the thermally destabilizing modification of the duplex in the seed region of the antisense strand includes nucleotides with impaired W-C H-bonding to complementary base on the target mRNA, such as: 
     
       
         
         
             
             
         
       
     
     More examples of abasic nucleotide, acyclic nucleotide modifications (including UNA and GNA), and mismatch modifications have been described in detail in WO 2011/133876, which is herein incorporated by reference in its entirety. 
     The thermally destabilizing modifications may also include universal base with reduced or abolished capability to form hydrogen bonds with the opposing bases, and phosphate modifications. 
     In some embodiments, the thermally destabilizing modification of the duplex includes nucleotides with non-canonical bases such as, but not limited to, nucleobase modifications with impaired or completely abolished capability to form hydrogen bonds with bases in the opposite strand. These nucleobase modifications have been evaluated for destabilization of the central region of the dsRNA duplex as described in WO 2010/0011895, which is herein incorporated by reference in its entirety. Exemplary nucleobase modifications are: 
     
       
         
         
             
             
         
       
     
     In some embodiments, the thermally destabilizing modification of the duplex in the seed region of the antisense strand includes one or more α-nucleotide complementary to the base on the target mRNA, such as: 
     
       
         
         
             
             
         
       
     
     wherein R is H, OH, OCH3, F, NH 2 , NHMe, NMe 2  or O-alkyl. 
     Exemplary phosphate modifications known to decrease the thermal stability of dsRNA duplexes compared to natural phosphodiester linkages are: 
     
       
         
         
             
             
         
       
     
     The alkyl for the R group can be a C 1 -C 6 alkyl. Specific alkyls for the R group include, but are not limited to methyl, ethyl, propyl, isopropyl, butyl, pentyl and hexyl. 
     As the skilled artisan will recognize, in view of the functional role of nucleobases is defining specificity of a RNAi agent of the disclosure, while nucleobase modifications can be performed in the various manners as described herein, e.g., to introduce destabilizing modifications into a RNAi agent of the disclosure, e.g., for purpose of enhancing on-target effect relative to off-target effect, the range of modifications available and, in general, present upon RNAi agents of the disclosure tends to be much greater for non-nucleobase modifications, e.g., modifications to sugar groups or phosphate backbones of polyribonucleotides. Such modifications are described in greater detail in other sections of the instant disclosure and are expressly contemplated for RNAi agents of the disclosure, either possessing native nucleobases or modified nucleobases as described above or elsewhere herein. 
     In addition to the antisense strand comprising a thermally destabilizing modification, the dsRNA can also comprise one or more stabilizing modifications. For example, the dsRNA can comprise at least two (e.g., two, three, four, five, six, seven, eight, nine, ten or more) stabilizing modifications. Without limitations, the stabilizing modifications all can be present in one strand. In some embodiments, both the sense and the antisense strands comprise at least two stabilizing modifications. The stabilizing modification can occur on any nucleotide of the sense strand or antisense strand. For instance, the stabilizing modification can occur on every nucleotide on the sense strand or antisense strand; each stabilizing modification can occur in an alternating pattern on the sense strand or antisense strand; or the sense strand or antisense strand comprises both stabilizing modification in an alternating pattern. The alternating pattern of the stabilizing modifications on the sense strand may be the same or different from the antisense strand, and the alternating pattern of the stabilizing modifications on the sense strand can have a shift relative to the alternating pattern of the stabilizing modifications on the antisense strand. 
     In some embodiments, the antisense strand comprises at least two (e.g., two, three, four, five, six, seven, eight, nine, ten or more) stabilizing modifications. Without limitations, a stabilizing modification in the antisense strand can be present at any positions. In some embodiments, the antisense comprises stabilizing modifications at positions 2, 6, 8, 9, 14, and 16 from the 5′-end. In some other embodiments, the antisense comprises stabilizing modifications at positions 2, 6, 14, and 16 from the 5′-end. In still some other embodiments, the antisense comprises stabilizing modifications at positions 2, 14, and 16 from the 5′-end. 
     In some embodiments, the antisense strand comprises at least one stabilizing modification adjacent to the destabilizing modification. For example, the stabilizing modification can be the nucleotide at the 5′-end or the 3′-end of the destabilizing modification, i.e., at position −1 or +1 from the position of the destabilizing modification. In some embodiments, the antisense strand comprises a stabilizing modification at each of the 5′-end and the 3′-end of the destabilizing modification, i.e., positions −1 and +1 from the position of the destabilizing modification. 
     In some embodiments, the antisense strand comprises at least two stabilizing modifications at the 3′-end of the destabilizing modification, i.e., at positions +1 and +2 from the position of the destabilizing modification. 
     In some embodiments, the sense strand comprises at least two (e.g., two, three, four, five, six, seven, eight, nine, ten or more) stabilizing modifications. Without limitations, a stabilizing modification in the sense strand can be present at any positions. In some embodiments, the sense strand comprises stabilizing modifications at positions 7, 10, and 11 from the 5′-end. In some other embodiments, the sense strand comprises stabilizing modifications at positions 7, 9, 10, and 11 from the 5′-end. In some embodiments, the sense strand comprises stabilizing modifications at positions opposite or complimentary to positions 11, 12, and 15 of the antisense strand, counting from the 5′-end of the antisense strand. In some other embodiments, the sense strand comprises stabilizing modifications at positions opposite or complimentary to positions 11, 12, 13, and 15 of the antisense strand, counting from the 5′-end of the antisense strand. In some embodiments, the sense strand comprises a block of two, three, or four stabilizing modifications. 
     In some embodiments, the sense strand does not comprise a stabilizing modification in position opposite or complimentary to the thermally destabilizing modification of the duplex in the antisense strand. 
     Exemplary thermally stabilizing modifications include, but are not limited to, 2′-fluoro modifications. Other thermally stabilizing modifications include, but are not limited to, LNA. 
     In some embodiments, the dsRNA of the disclosure comprises at least four (e.g., four, five, six, seven, eight, nine, ten, or more) 2′-fluoro nucleotides. Without limitations, the 2′-fluoro nucleotides all can be present in one strand. In some embodiments, both the sense and the antisense strands comprise at least two 2′-fluoro nucleotides. The 2′-fluoro modification can occur on any nucleotide of the sense strand or antisense strand. For instance, the 2′-fluoro modification can occur on every nucleotide on the sense strand or antisense strand; each 2′-fluoro modification can occur in an alternating pattern on the sense strand or antisense strand; or the sense strand or antisense strand comprises both 2′-fluoro modifications in an alternating pattern. The alternating pattern of the 2′-fluoro modifications on the sense strand may be the same or different from the antisense strand, and the alternating pattern of the 2′-fluoro modifications on the sense strand can have a shift relative to the alternating pattern of the 2′-fluoro modifications on the antisense strand. 
     In some embodiments, the antisense strand comprises at least two (e.g., two, three, four, five, six, seven, eight, nine, ten, or more) 2′-fluoro nucleotides. Without limitations, a 2′-fluoro modification in the antisense strand can be present at any positions. In some embodiments, the antisense comprises 2′-fluoro nucleotides at positions 2, 6, 8, 9, 14, and 16 from the 5′-end. In some other embodiments, the antisense comprises 2′-fluoro nucleotides at positions 2, 6, 14, and 16 from the 5′-end. In still some other embodiments, the antisense comprises 2′-fluoro nucleotides at positions 2, 14, and 16 from the 5′-end. 
     In some embodiments, the antisense strand comprises at least one 2′-fluoro nucleotide adjacent to the destabilizing modification. For example, the 2′-fluoro nucleotide can be the nucleotide at the 5′-end or the 3′-end of the destabilizing modification, i.e., at position −1 or +1 from the position of the destabilizing modification. In some embodiments, the antisense strand comprises a 2′-fluoro nucleotide at each of the 5′-end and the 3′-end of the destabilizing modification, i.e., positions −1 and +1 from the position of the destabilizing modification. 
     In some embodiments, the antisense strand comprises at least two 2′-fluoro nucleotides at the 3′-end of the destabilizing modification, i.e., at positions +1 and +2 from the position of the destabilizing modification. 
     In some embodiments, the sense strand comprises at least two (e.g., two, three, four, five, six, seven, eight, nine, ten, or more) 2′-fluoro nucleotides. Without limitations, a 2′-fluoro modification in the sense strand can be present at any positions. In some embodiments, the antisense comprises 2′-fluoro nucleotides at positions 7, 10, and 11 from the 5′-end. In some other embodiments, the sense strand comprises 2′-fluoro nucleotides at positions 7, 9, 10, and 11 from the 5′-end. In some embodiments, the sense strand comprises 2′-fluoro nucleotides at positions opposite or complimentary to positions 11, 12, and 15 of the antisense strand, counting from the 5′-end of the antisense strand. In some other embodiments, the sense strand comprises 2′-fluoro nucleotides at positions opposite or complimentary to positions 11, 12, 13, and 15 of the antisense strand, counting from the 5′-end of the antisense strand. In some embodiments, the sense strand comprises a block of two, three or four 2′-fluoro nucleotides. 
     In some embodiments, the sense strand does not comprise a 2′-fluoro nucleotide in position opposite or complimentary to the thermally destabilizing modification of the duplex in the antisense strand. 
     In some embodiments, the dsRNA molecule of the disclosure comprises a 21 nucleotides (nt) sense strand and a 23 nucleotides (nt) antisense, wherein the antisense strand contains at least one thermally destabilizing nucleotide, where the at least one thermally destabilizing nucleotide occurs in the seed region of the antisense strand (i.e., at position 2-9 of the 5′-end of the antisense strand), wherein one end of the dsRNA is blunt, while the other end is comprises a 2 nt overhang, and wherein the dsRNA optionally further has at least one (e.g., one, two, three, four, five, six or all seven) of the following characteristics: (i) the antisense comprises 2, 3, 4, 5, or 6 2′-fluoro modifications; (ii) the antisense comprises 1, 2, 3, 4, or 5 phosphorothioate internucleotide linkages; (iii) the sense strand is conjugated with a ligand; (iv) the sense strand comprises 2, 3, 4, or 5 2′-fluoro modifications; (v) the sense strand comprises 1, 2, 3, 4, or 5 phosphorothioate internucleotide linkages; (vi) the dsRNA comprises at least four 2′-fluoro modifications; and (vii) the dsRNA comprises a blunt end at 5′-end of the antisense strand. Preferably, the 2 nt overhang is at the 3′-end of the antisense. 
     In some embodiments, the dsRNA molecule of the disclosure comprising a sense and antisense strands, wherein: the sense strand is 25-30 nucleotide residues in length, wherein starting from the 5′ terminal nucleotide (position 1), positions 1 to 23 of said sense strand comprise at least 8 ribonucleotides; antisense strand is 36-66 nucleotide residues in length and, starting from the 3′ terminal nucleotide, at least 8 ribonucleotides in the positions paired with positions 1-23 of sense strand to form a duplex; wherein at least the 3 ‘ terminal nucleotide of antisense strand is unpaired with sense strand, and up to 6 consecutive 3’ terminal nucleotides are unpaired with sense strand, thereby forming a 3′ single stranded overhang of 1-6 nucleotides; wherein the 5′ terminus of antisense strand comprises from 10-30 consecutive nucleotides which are unpaired with sense strand, thereby forming a 10-30 nucleotide single stranded 5′ overhang; wherein at least the sense strand 5′ terminal and 3′ terminal nucleotides are base paired with nucleotides of antisense strand when sense and antisense strands are aligned for maximum complementarity, thereby forming a substantially duplexed region between sense and antisense strands; and antisense strand is sufficiently complementary to a target RNA along at least 19 ribonucleotides of antisense strand length to reduce target gene expression when said double stranded nucleic acid is introduced into a mammalian cell; and wherein the antisense strand contains at least one thermally destabilizing nucleotide, where at least one thermally destabilizing nucleotide is in the seed region of the antisense strand (i.e. at position 2-9 of the 5′-end of the antisense strand). For example, the thermally destabilizing nucleotide occurs between positions opposite or complimentary to positions 14-17 of the 5′-end of the sense strand, and wherein the dsRNA optionally further has at least one (e.g., one, two, three, four, five, six or all seven) of the following characteristics: (i) the antisense comprises 2, 3, 4, 5, or 6 2′-fluoro modifications; (ii) the antisense comprises 1, 2, 3, 4, or 5 phosphorothioate internucleotide linkages; (iii) the sense strand is conjugated with a ligand; (iv) the sense strand comprises 2, 3, 4, or 5 2′-fluoro modifications; (v) the sense strand comprises 1, 2, 3, 4, or 5 phosphorothioate internucleotide linkages; and (vi) the dsRNA comprises at least four 2′-fluoro modifications; and (vii) the dsRNA comprises a duplex region of 12-30 nucleotide pairs in length. 
     In some embodiments, the dsRNA molecule of the disclosure comprises a sense and antisense strands, wherein said dsRNA molecule comprises a sense strand having a length which is at least 25 and at most 29 nucleotides and an antisense strand having a length which is at most 30 nucleotides with the sense strand comprises a modified nucleotide that is susceptible to enzymatic degradation at position 11 from the 5′end, wherein the 3′ end of said sense strand and the 5′ end of said antisense strand form a blunt end and said antisense strand is 1˜4 nucleotides longer at its 3′ end than the sense strand, wherein the duplex region which is at least 25 nucleotides in length, and said antisense strand is sufficiently complementary to a target mRNA along at least 19 nt of said antisense strand length to reduce target gene expression when said dsRNA molecule is introduced into a mammalian cell, and wherein dicer cleavage of said dsRNA preferentially results in an siRNA comprising said 3′ end of said antisense strand, thereby reducing expression of the target gene in the mammal, wherein the antisense strand contains at least one thermally destabilizing nucleotide, where the at least one thermally destabilizing nucleotide is in the seed region of the antisense strand (i.e. at position 2-9 of the 5′-end of the antisense strand), and wherein the dsRNA optionally further has at least one (e.g., one, two, three, four, five, six or all seven) of the following characteristics: (i) the antisense comprises 2, 3, 4, 5, or 6 2′-fluoro modifications; (ii) the antisense comprises 1, 2, 3, 4, or 5 phosphorothioate internucleotide linkages; (iii) the sense strand is conjugated with a ligand; (iv) the sense strand comprises 2, 3, 4, or 5 2′-fluoro modifications; (v) the sense strand comprises 1, 2, 3, 4, or 5 phosphorothioate internucleotide linkages; and (vi) the dsRNA comprises at least four 2′-fluoro modifications; and (vii) the dsRNA has a duplex region of 12-29 nucleotide pairs in length. 
     In some embodiments, every nucleotide in the sense strand and antisense strand of the dsRNA molecule may be modified. Each nucleotide may be modified with the same or different modification which can include one or more alteration of one or both of the non-linking phosphate oxygens or of one or more of the linking phosphate oxygens; alteration of a constituent of the ribose sugar, e.g., of the 2′ hydroxyl on the ribose sugar; wholesale replacement of the phosphate moiety with “dephospho” linkers; modification or replacement of a naturally occurring base; and replacement or modification of the ribose-phosphate backbone. 
     As nucleic acids are polymers of subunits, many of the modifications occur at a position which is repeated within a nucleic acid, e.g., a modification of a base, or a phosphate moiety, or a non-linking O of a phosphate moiety. In some cases, the modification will occur at all of the subject positions in the nucleic acid but in many cases it will not. By way of example, a modification may only occur at a 3′ or 5′ terminal position, may only occur in a terminal region, e.g., at a position on a terminal nucleotide or in the last 2, 3, 4, 5, or 10 nucleotides of a strand. A modification may occur in a double strand region, a single strand region, or in both. A modification may occur only in the double strand region of an RNA or may only occur in a single strand region of an RNA. e.g., a phosphorothioate modification at a non-linking O position may only occur at one or both termini, may only occur in a terminal region, e.g., at a position on a terminal nucleotide or in the last 2, 3, 4, 5, or 10 nucleotides of a strand, or may occur in double strand and single strand regions, particularly at termini. The 5′ end or ends can be phosphorylated. 
     It may be possible, e.g., to enhance stability, to include particular bases in overhangs, or to include modified nucleotides or nucleotide surrogates, in single strand overhangs, e.g., in a 5′ or 3′ overhang, or in both. E.g., it can be desirable to include purine nucleotides in overhangs. In some embodiments all or some of the bases in a 3′ or 5′ overhang may be modified, e.g., with a modification described herein. Modifications can include, e.g., the use of modifications at the 2′ position of the ribose sugar with modifications that are known in the art, e.g., the use of deoxyribonucleotides, 2′-deoxy-2′-fluoro (2′-F) or 2′-O-methyl modified instead of the ribosugar of the nucleobase, and modifications in the phosphate group, e.g., phosphorothioate modifications. Overhangs need not be homologous with the target sequence. 
     In some embodiments, each residue of the sense strand and antisense strand is independently modified with LNA, HNA, CeNA, 2′-methoxyethyl, 2′-O-methyl, 2′-O-allyl, 2′-C-allyl, 2′-deoxy, or 2′-fluoro. The strands can contain more than one modification. In some embodiments, each residue of the sense strand and antisense strand is independently modified with 2′-O-methyl or 2′-fluoro. It is to be understood that these modifications are in addition to the at least one thermally destabilizing modification of the duplex present in the antisense strand. 
     At least two different modifications are typically present on the sense strand and antisense strand. Those two modifications may be the 2′-deoxy, 2′-O-methyl or 2′-fluoro modifications, acyclic nucleotides or others. In some embodiments, the sense strand and antisense strand each comprises two differently modified nucleotides selected from 2′-O-methyl or 2′-deoxy. In some embodiments, each residue of the sense strand and antisense strand is independently modified with 2′-O-methyl nucleotide, 2′-deoxy nucleotide, 2′-deoxy-2′-fluoro nucleotide, 2′-O—N-methylacetamido (2′-O-NMA) nucleotide, a 2′-O-dimethylaminoethoxyethyl (2′-O-DMAEOE) nucleotide, 2′ aminopropyl (2′-O-AP) nucleotide, or 2′-ara-F nucleotide. Again, it is to be understood that these modifications are in addition to the at least one thermally destabilizing modification of the duplex present in the antisense strand. 
     In some embodiments, the dsRNA molecule of the disclosure comprises modifications of an alternating pattern, particular in the B1, B2, B3, B1′, B2′, B3′, B4′ regions. The term “alternating motif” or “alternative pattern” as used herein refers to a motif having one or more modifications, each modification occurring on alternating nucleotides of one strand. The alternating nucleotide may refer to one per every other nucleotide or one per every three nucleotides, or a similar pattern. For example, if A, B and C each represent one type of modification to the nucleotide, the alternating motif can be “ABABABABABAB . . . ,” “AABBAABBAABB . . . ,” “AABAABAABAAB . . . ,” “AAABAAABAAAB . . . ,” “AAABBBAAABBB . . . ,” or “ABCABCABCABC . . . ,” etc. 
     The type of modifications contained in the alternating motif may be the same or different. For example, if A, B, C, D each represent one type of modification on the nucleotide, the alternating pattern, i.e., modifications on every other nucleotide, may be the same, but each of the sense strand or antisense strand can be selected from several possibilities of modifications within the alternating motif such as “ABABAB . . . ”, “ACACAC . . . ” “BDBDBD . . . ” or “CDCDCD . . . ,” etc. 
     In some embodiments, the dsRNA molecule of the disclosure comprises the modification pattern for the alternating motif on the sense strand relative to the modification pattern for the alternating motif on the antisense strand is shifted. The shift may be such that the modified group of nucleotides of the sense strand corresponds to a differently modified group of nucleotides of the antisense strand and vice versa. For example, the sense strand when paired with the antisense strand in the dsRNA duplex, the alternating motif in the sense strand may start with “ABABAB” from 5′-3′ of the strand and the alternating motif in the antisense strand may start with “BABABA” from 3′-5′ of the strand within the duplex region. As another example, the alternating motif in the sense strand may start with “AABBAABB” from 5′-3′ of the strand and the alternating motif in the antisense strand may start with “BBAABBAA” from 3′-5′ of the strand within the duplex region, so that there is a complete or partial shift of the modification patterns between the sense strand and the antisense strand. 
     The dsRNA molecule of the disclosure may further comprise at least one phosphorothioate or methylphosphonate internucleotide linkage. The phosphorothioate or methylphosphonate internucleotide linkage modification may occur on any nucleotide of the sense strand or antisense strand or both in any position of the strand. For instance, the internucleotide linkage modification may occur on every nucleotide on the sense strand or antisense strand; each internucleotide linkage modification may occur in an alternating pattern on the sense strand or antisense strand; or the sense strand or antisense strand comprises both internucleotide linkage modifications in an alternating pattern. The alternating pattern of the internucleotide linkage modification on the sense strand may be the same or different from the antisense strand, and the alternating pattern of the internucleotide linkage modification on the sense strand may have a shift relative to the alternating pattern of the internucleotide linkage modification on the antisense strand. 
     In some embodiments, the dsRNA molecule comprises the phosphorothioate or methylphosphonate internucleotide linkage modification in the overhang region. For example, the overhang region comprises two nucleotides having a phosphorothioate or methylphosphonate internucleotide linkage between the two nucleotides. Internucleotide linkage modifications also may be made to link the overhang nucleotides with the terminal paired nucleotides within duplex region. For example, at least 2, 3, 4, or all the overhang nucleotides may be linked through phosphorothioate or methylphosphonate internucleotide linkage, and optionally, there may be additional phosphorothioate or methylphosphonate internucleotide linkages linking the overhang nucleotide with a paired nucleotide that is next to the overhang nucleotide. For instance, there may be at least two phosphorothioate internucleotide linkages between the terminal three nucleotides, in which two of the three nucleotides are overhang nucleotides, and the third is a paired nucleotide next to the overhang nucleotide. Preferably, these terminal three nucleotides may be at the 3′-end of the antisense strand. 
     In some embodiments, the sense strand of the dsRNA molecule comprises 1-10 blocks of two to ten phosphorothioate or methylphosphonate internucleotide linkages separated by 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, or 16 phosphate internucleotide linkages, wherein one of the phosphorothioate or methylphosphonate internucleotide linkages is placed at any position in the oligonucleotide sequence and the said sense strand is paired with an antisense strand comprising any combination of phosphorothioate, methylphosphonate and phosphate internucleotide linkages or an antisense strand comprising either phosphorothioate or methylphosphonate or phosphate linkage. 
     In some embodiments, the antisense strand of the dsRNA molecule comprises two blocks of two phosphorothioate or methylphosphonate internucleotide linkages separated by 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, or 18 phosphate internucleotide linkages, wherein one of the phosphorothioate or methylphosphonate internucleotide linkages is placed at any position in the oligonucleotide sequence and the said antisense strand is paired with a sense strand comprising any combination of phosphorothioate, methylphosphonate and phosphate internucleotide linkages or an antisense strand comprising either phosphorothioate or methylphosphonate or phosphate linkage. 
     In some embodiments, the antisense strand of the dsRNA molecule comprises two blocks of three phosphorothioate or methylphosphonate internucleotide linkages separated by 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, or 16 phosphate internucleotide linkages, wherein one of the phosphorothioate or methylphosphonate internucleotide linkages is placed at any position in the oligonucleotide sequence and the said antisense strand is paired with a sense strand comprising any combination of phosphorothioate, methylphosphonate and phosphate internucleotide linkages or an antisense strand comprising either phosphorothioate or methylphosphonate or phosphate linkage. 
     In some embodiments, the antisense strand of the dsRNA molecule comprises two blocks of four phosphorothioate or methylphosphonate internucleotide linkages separated by 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, or 14 phosphate internucleotide linkages, wherein one of the phosphorothioate or methylphosphonate internucleotide linkages is placed at any position in the oligonucleotide sequence and the said antisense strand is paired with a sense strand comprising any combination of phosphorothioate, methylphosphonate and phosphate internucleotide linkages or an antisense strand comprising either phosphorothioate or methylphosphonate or phosphate linkage. 
     In some embodiments, the antisense strand of the dsRNA molecule comprises two blocks of five phosphorothioate or methylphosphonate internucleotide linkages separated by 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12 phosphate internucleotide linkages, wherein one of the phosphorothioate or methylphosphonate internucleotide linkages is placed at any position in the oligonucleotide sequence and the said antisense strand is paired with a sense strand comprising any combination of phosphorothioate, methylphosphonate and phosphate internucleotide linkages or an antisense strand comprising either phosphorothioate or methylphosphonate or phosphate linkage. 
     In some embodiments, the antisense strand of the dsRNA molecule comprises two blocks of six phosphorothioate or methylphosphonate internucleotide linkages separated by 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 phosphate internucleotide linkages, wherein one of the phosphorothioate or methylphosphonate internucleotide linkages is placed at any position in the oligonucleotide sequence and the said antisense strand is paired with a sense strand comprising any combination of phosphorothioate, methylphosphonate and phosphate internucleotide linkages or an antisense strand comprising either phosphorothioate or methylphosphonate or phosphate linkage. 
     In some embodiments, the antisense strand of the dsRNA molecule comprises two blocks of seven phosphorothioate or methylphosphonate internucleotide linkages separated by 1, 2, 3, 4, 5, 6, 7, or 8 phosphate internucleotide linkages, wherein one of the phosphorothioate or methylphosphonate internucleotide linkages is placed at any position in the oligonucleotide sequence and the said antisense strand is paired with a sense strand comprising any combination of phosphorothioate, methylphosphonate and phosphate internucleotide linkages or an antisense strand comprising either phosphorothioate or methylphosphonate or phosphate linkage. 
     In some embodiments, the antisense strand of the dsRNA molecule comprises two blocks of eight phosphorothioate or methylphosphonate internucleotide linkages separated by 1, 2, 3, 4, 5, or 6 phosphate internucleotide linkages, wherein one of the phosphorothioate or methylphosphonate internucleotide linkages is placed at any position in the oligonucleotide sequence and the said antisense strand is paired with a sense strand comprising any combination of phosphorothioate, methylphosphonate and phosphate internucleotide linkages or an antisense strand comprising either phosphorothioate or methylphosphonate or phosphate linkage. 
     In some embodiments, the antisense strand of the dsRNA molecule comprises two blocks of nine phosphorothioate or methylphosphonate internucleotide linkages separated by 1, 2, 3, or 4 phosphate internucleotide linkages, wherein one of the phosphorothioate or methylphosphonate internucleotide linkages is placed at any position in the oligonucleotide sequence and the said antisense strand is paired with a sense strand comprising any combination of phosphorothioate, methylphosphonate and phosphate internucleotide linkages or an antisense strand comprising either phosphorothioate or methylphosphonate or phosphate linkage. 
     In some embodiments, the dsRNA molecule of the disclosure further comprises one or more phosphorothioate or methylphosphonate internucleotide linkage modification within 1-10 of the termini position(s) of the sense or antisense strand. For example, at least 2, 3, 4, 5, 6, 7, 8, 9, or 10 nucleotides may be linked through phosphorothioate or methylphosphonate internucleotide linkage at one end or both ends of the sense or antisense strand. 
     In some embodiments, the dsRNA molecule of the disclosure further comprises one or more phosphorothioate or methylphosphonate internucleotide linkage modification within 1-10 of the internal region of the duplex of each of the sense or antisense strand. For example, at least 2, 3, 4, 5, 6, 7, 8, 9, or 10 nucleotides may be linked through phosphorothioate methylphosphonate internucleotide linkage at position 8-16 of the duplex region counting from the 5′-end of the sense strand; the dsRNA molecule can optionally further comprise one or more phosphorothioate or methylphosphonate internucleotide linkage modification within 1-10 of the termini position(s). 
     In some embodiments, the dsRNA molecule of the disclosure further comprises one to five phosphorothioate or methylphosphonate internucleotide linkage modification(s) within position 1-5 and one to five phosphorothioate or methylphosphonate internucleotide linkage modification(s) within position 18-23 of the sense strand (counting from the 5′-end), and one to five phosphorothioate or methylphosphonate internucleotide linkage modification at positions 1 and 2 and one to five within positions 18-23 of the antisense strand (counting from the 5′-end). 
     In some embodiments, the dsRNA molecule of the disclosure further comprises one phosphorothioate internucleotide linkage modification within position 1-5 and one phosphorothioate or methylphosphonate internucleotide linkage modification within position 18-23 of the sense strand (counting from the 5′-end), and one phosphorothioate internucleotide linkage modification at positions 1 and 2 and two phosphorothioate or methylphosphonate internucleotide linkage modifications within positions 18-23 of the antisense strand (counting from the 5′-end). 
     In some embodiments, the dsRNA molecule of the disclosure further comprises two phosphorothioate internucleotide linkage modifications within position 1-5 and one phosphorothioate internucleotide linkage modification within position 18-23 of the sense strand (counting from the 5′-end), and one phosphorothioate internucleotide linkage modification at positions 1 and 2 and two phosphorothioate internucleotide linkage modifications within positions 18-23 of the antisense strand (counting from the 5′-end). 
     In some embodiments, the dsRNA molecule of the disclosure further comprises two phosphorothioate internucleotide linkage modifications within position 1-5 and two phosphorothioate internucleotide linkage modifications within position 18-23 of the sense strand (counting from the 5′-end), and one phosphorothioate internucleotide linkage modification at positions 1 and 2 and two phosphorothioate internucleotide linkage modifications within positions 18-23 of the antisense strand (counting from the 5′-end). 
     In some embodiments, the dsRNA molecule of the disclosure further comprises two phosphorothioate internucleotide linkage modifications within position 1-5 and two phosphorothioate internucleotide linkage modifications within position 18-23 of the sense strand (counting from the 5′-end), and one phosphorothioate internucleotide linkage modification at positions 1 and 2 and one phosphorothioate internucleotide linkage modification within positions 18-23 of the antisense strand (counting from the 5′-end). 
     In some embodiments, the dsRNA molecule of the disclosure further comprises one phosphorothioate internucleotide linkage modification within position 1-5 and one phosphorothioate internucleotide linkage modification within position 18-23 of the sense strand (counting from the 5′-end), and two phosphorothioate internucleotide linkage modifications at positions 1 and 2 and two phosphorothioate internucleotide linkage modifications within positions 18-23 of the antisense strand (counting from the 5′-end). 
     In some embodiments, the dsRNA molecule of the disclosure further comprises one phosphorothioate internucleotide linkage modification within position 1-5 and one within position 18-23 of the sense strand (counting from the 5′-end), and two phosphorothioate internucleotide linkage modification at positions 1 and 2 and one phosphorothioate internucleotide linkage modification within positions 18-23 of the antisense strand (counting from the 5′-end). 
     In some embodiments, the dsRNA molecule of the disclosure further comprises one phosphorothioate internucleotide linkage modification within position 1-5 (counting from the 5′-end) of the sense strand, and two phosphorothioate internucleotide linkage modifications at positions 1 and 2 and one phosphorothioate internucleotide linkage modification within positions 18-23 of the antisense strand (counting from the 5′-end). 
     In some embodiments, the dsRNA molecule of the disclosure further comprises two phosphorothioate internucleotide linkage modifications within position 1-5 (counting from the 5′-end) of the sense strand, and one phosphorothioate internucleotide linkage modification at positions 1 and 2 and two phosphorothioate internucleotide linkage modifications within positions 18-23 of the antisense strand (counting from the 5′-end). 
     In some embodiments, the dsRNA molecule of the disclosure further comprises two phosphorothioate internucleotide linkage modifications within position 1-5 and one within position 18-23 of the sense strand (counting from the 5′-end), and two phosphorothioate internucleotide linkage modifications at positions 1 and 2 and one phosphorothioate internucleotide linkage modification within positions 18-23 of the antisense strand (counting from the 5′-end). 
     In some embodiments, the dsRNA molecule of the disclosure further comprises two phosphorothioate internucleotide linkage modifications within position 1-5 and one phosphorothioate internucleotide linkage modification within position 18-23 of the sense strand (counting from the 5′-end), and two phosphorothioate internucleotide linkage modifications at positions 1 and 2 and two phosphorothioate internucleotide linkage modifications within positions 18-23 of the antisense strand (counting from the 5′-end). 
     In some embodiments, the dsRNA molecule of the disclosure further comprises two phosphorothioate internucleotide linkage modifications within position 1-5 and one phosphorothioate internucleotide linkage modification within position 18-23 of the sense strand (counting from the 5′-end), and one phosphorothioate internucleotide linkage modification at positions 1 and 2 and two phosphorothioate internucleotide linkage modifications within positions 18-23 of the antisense strand (counting from the 5′-end). 
     In some embodiments, the dsRNA molecule of the disclosure further comprises two phosphorothioate internucleotide linkage modifications at position 1 and 2, and two phosphorothioate internucleotide linkage modifications at position 20 and 21 of the sense strand (counting from the 5′-end), and one phosphorothioate internucleotide linkage modification at positions 1 and one at position 21 of the antisense strand (counting from the 5′-end). 
     In some embodiments, the dsRNA molecule of the disclosure further comprises one phosphorothioate internucleotide linkage modification at position 1, and one phosphorothioate internucleotide linkage modification at position 21 of the sense strand (counting from the 5′-end), and two phosphorothioate internucleotide linkage modifications at positions 1 and 2 and two phosphorothioate internucleotide linkage modifications at positions 20 and 21 the antisense strand (counting from the 5′-end). 
     In some embodiments, the dsRNA molecule of the disclosure further comprises two phosphorothioate internucleotide linkage modifications at position 1 and 2, and two phosphorothioate internucleotide linkage modifications at position 21 and 22 of the sense strand (counting from the 5′-end), and one phosphorothioate internucleotide linkage modification at positions 1 and one phosphorothioate internucleotide linkage modification at position 21 of the antisense strand (counting from the 5′-end). 
     In some embodiments, the dsRNA molecule of the disclosure further comprises one phosphorothioate internucleotide linkage modification at position 1, and one phosphorothioate internucleotide linkage modification at position 21 of the sense strand (counting from the 5′-end), and two phosphorothioate internucleotide linkage modifications at positions 1 and 2 and two phosphorothioate internucleotide linkage modifications at positions 21 and 22 the antisense strand (counting from the 5′-end). 
     In some embodiments, the dsRNA molecule of the disclosure further comprises two phosphorothioate internucleotide linkage modifications at position 1 and 2, and two phosphorothioate internucleotide linkage modifications at position 22 and 23 of the sense strand (counting from the 5′-end), and one phosphorothioate internucleotide linkage modification at positions 1 and one phosphorothioate internucleotide linkage modification at position 21 of the antisense strand (counting from the 5′-end). 
     In some embodiments, the dsRNA molecule of the disclosure further comprises one phosphorothioate internucleotide linkage modification at position 1, and one phosphorothioate internucleotide linkage modification at position 21 of the sense strand (counting from the 5′-end), and two phosphorothioate internucleotide linkage modifications at positions 1 and 2 and two phosphorothioate internucleotide linkage modifications at positions 23 and 23 the antisense strand (counting from the 5′-end). 
     In some embodiments, compound of the disclosure comprises a pattern of backbone chiral centers. In some embodiments, a common pattern of backbone chiral centers comprises at least 5 internucleotidic linkages in the Sp configuration. In some embodiments, a common pattern of backbone chiral centers comprises at least 6 internucleotidic linkages in the Sp configuration. In some embodiments, a common pattern of backbone chiral centers comprises at least 7 internucleotidic linkages in the Sp configuration. In some embodiments, a common pattern of backbone chiral centers comprises at least 8 internucleotidic linkages in the Sp configuration. In some embodiments, a common pattern of backbone chiral centers comprises at least 9 internucleotidic linkages in the Sp configuration. In some embodiments, a common pattern of backbone chiral centers comprises at least 10 internucleotidic linkages in the Sp configuration. In some embodiments, a common pattern of backbone chiral centers comprises at least 11 internucleotidic linkages in the Sp configuration. In some embodiments, a common pattern of backbone chiral centers comprises at least 12 internucleotidic linkages in the Sp configuration. In some embodiments, a common pattern of backbone chiral centers comprises at least 13 internucleotidic linkages in the Sp configuration. In some embodiments, a common pattern of backbone chiral centers comprises at least 14 internucleotidic linkages in the Sp configuration. In some embodiments, a common pattern of backbone chiral centers comprises at least 15 internucleotidic linkages in the Sp configuration. In some embodiments, a common pattern of backbone chiral centers comprises at least 16 internucleotidic linkages in the Sp configuration. In some embodiments, a common pattern of backbone chiral centers comprises at least 17 internucleotidic linkages in the Sp configuration. In some embodiments, a common pattern of backbone chiral centers comprises at least 18 internucleotidic linkages in the Sp configuration. In some embodiments, a common pattern of backbone chiral centers comprises at least 19 internucleotidic linkages in the Sp configuration. In some embodiments, a common pattern of backbone chiral centers comprises no more than 8 internucleotidic linkages in the Rp configuration. In some embodiments, a common pattern of backbone chiral centers comprises no more than 7 internucleotidic linkages in the Rp configuration. In some embodiments, a common pattern of backbone chiral centers comprises no more than 6 internucleotidic linkages in the Rp configuration. In some embodiments, a common pattern of backbone chiral centers comprises no more than 5 internucleotidic linkages in the Rp configuration. In some embodiments, a common pattern of backbone chiral centers comprises no more than 4 internucleotidic linkages in the Rp configuration. In some embodiments, a common pattern of backbone chiral centers comprises no more than 3 internucleotidic linkages in the Rp configuration. In some embodiments, a common pattern of backbone chiral centers comprises no more than 2 internucleotidic linkages in the Rp configuration. In some embodiments, a common pattern of backbone chiral centers comprises no more than 1 internucleotidic linkages in the Rp configuration. In some embodiments, a common pattern of backbone chiral centers comprises no more than 8 internucleotidic linkages which are not chiral (as a non-limiting example, a phosphodiester). In some embodiments, a common pattern of backbone chiral centers comprises no more than 7 internucleotidic linkages which are not chiral. In some embodiments, a common pattern of backbone chiral centers comprises no more than 6 internucleotidic linkages which are not chiral. In some embodiments, a common pattern of backbone chiral centers comprises no more than 5 internucleotidic linkages which are not chiral. In some embodiments, a common pattern of backbone chiral centers comprises no more than 4 internucleotidic linkages which are not chiral. In some embodiments, a common pattern of backbone chiral centers comprises no more than 3 internucleotidic linkages which are not chiral. In some embodiments, a common pattern of backbone chiral centers comprises no more than 2 internucleotidic linkages which are not chiral. In some embodiments, a common pattern of backbone chiral centers comprises no more than 1 internucleotidic linkages which are not chiral. In some embodiments, a common pattern of backbone chiral centers comprises at least 10 internucleotidic linkages in the Sp configuration, and no more than 8 internucleotidic linkages which are not chiral. In some embodiments, a common pattern of backbone chiral centers comprises at least 11 internucleotidic linkages in the Sp configuration, and no more than 7 internucleotidic linkages which are not chiral. In some embodiments, a common pattern of backbone chiral centers comprises at least 12 internucleotidic linkages in the Sp configuration, and no more than 6 internucleotidic linkages which are not chiral. In some embodiments, a common pattern of backbone chiral centers comprises at least 13 internucleotidic linkages in the Sp configuration, and no more than 6 internucleotidic linkages which are not chiral. In some embodiments, a common pattern of backbone chiral centers comprises at least 14 internucleotidic linkages in the Sp configuration, and no more than 5 internucleotidic linkages which are not chiral. In some embodiments, a common pattern of backbone chiral centers comprises at least 15 internucleotidic linkages in the Sp configuration, and no more than 4 internucleotidic linkages which are not chiral. In some embodiments, the internucleotidic linkages in the Sp configuration are optionally contiguous or not contiguous. In some embodiments, the internucleotidic linkages in the Rp configuration are optionally contiguous or not contiguous. In some embodiments, the internucleotidic linkages which are not chiral are optionally contiguous or not contiguous. 
     In some embodiments, compound of the disclosure comprises a block is a stereochemistry block. In some embodiments, a block is an Rp block in that each internucleotidic linkage of the block is Rp. In some embodiments, a 5′-block is an Rp block. In some embodiments, a 3′-block is an Rp block. In some embodiments, a block is an Sp block in that each internucleotidic linkage of the block is Sp. In some embodiments, a 5′-block is an Sp block. In some embodiments, a 3′-block is an Sp block. In some embodiments, provided oligonucleotides comprise both Rp and Sp blocks. In some embodiments, provided oligonucleotides comprise one or more Rp but no Sp blocks. In some embodiments, provided oligonucleotides comprise one or more Sp but no Rp blocks. In some embodiments, provided oligonucleotides comprise one or more PO blocks wherein each internucleotidic linkage in a natural phosphate linkage. 
     In some embodiments, compound of the disclosure comprises a 5′-block is an Sp block wherein each sugar moiety comprises a 2′-F modification. In some embodiments, a 5′-block is an Sp block wherein each of internucleotidic linkage is a modified internucleotidic linkage and each sugar moiety comprises a 2′-F modification. In some embodiments, a 5′-block is an Sp block wherein each of internucleotidic linkage is a phosphorothioate linkage and each sugar moiety comprises a 2′-F modification. In some embodiments, a 5′-block comprises 4 or more nucleoside units. In some embodiments, a 5′-block comprises 5 or more nucleoside units. In some embodiments, a 5′-block comprises 6 or more nucleoside units. In some embodiments, a 5′-block comprises 7 or more nucleoside units. In some embodiments, a 3′-block is an Sp block wherein each sugar moiety comprises a 2′-F modification. In some embodiments, a 3′-block is an Sp block wherein each of internucleotidic linkage is a modified internucleotidic linkage and each sugar moiety comprises a 2′-F modification. In some embodiments, a 3′-block is an Sp block wherein each of internucleotidic linkage is a phosphorothioate linkage and each sugar moiety comprises a 2′-F modification. In some embodiments, a 3′-block comprises 4 or more nucleoside units. In some embodiments, a 3′-block comprises 5 or more nucleoside units. In some embodiments, a 3′-block comprises 6 or more nucleoside units. In some embodiments, a 3′-block comprises 7 or more nucleoside units. 
     In some embodiments, compound of the disclosure comprises a type of nucleoside in a region or an oligonucleotide is followed by a specific type of internucleotidic linkage, e.g., natural phosphate linkage, modified internucleotidic linkage, Rp chiral internucleotidic linkage, Sp chiral internucleotidic linkage, etc. In some embodiments, A is followed by Sp. In some embodiments, A is followed by Rp. In some embodiments, A is followed by natural phosphate linkage (PO). In some embodiments, U is followed by Sp. In some embodiments, U is followed by Rp. In some embodiments, U is followed by natural phosphate linkage (PO). In some embodiments, C is followed by Sp. In some embodiments, C is followed by Rp. In some embodiments, C is followed by natural phosphate linkage (PO). In some embodiments, G is followed by Sp. In some embodiments, G is followed by Rp. In some embodiments, G is followed by natural phosphate linkage (PO). In some embodiments, C and U are followed by Sp. In some embodiments, C and U are followed by Rp. In some embodiments, C and U are followed by natural phosphate linkage (PO). In some embodiments, A and G are followed by Sp. In some embodiments, A and G are followed by Rp. 
     In some embodiments, the antisense strand comprises phosphorothioate internucleotide linkages between nucleotide positions 21 and 22, and between nucleotide positions 22 and 23, wherein the antisense strand contains at least one thermally destabilizing modification of the duplex located in the seed region of the antisense strand (i.e., at position 2-9 of the 5′-end of the antisense strand), and wherein the dsRNA optionally further has at least one (e.g., one, two, three, four, five, six, seven or all eight) of the following characteristics: (i) the antisense comprises 2, 3, 4, 5, or 6 2′-fluoro modifications; (ii) the antisense comprises 3, 4, or 5 phosphorothioate internucleotide linkages; (iii) the sense strand is conjugated with a ligand; (iv) the sense strand comprises 2, 3, 4 or 5 2′-fluoro modifications; (v) the sense strand comprises 1, 2, 3, 4, or 5 phosphorothioate internucleotide linkages; (vi) the dsRNA comprises at least four 2′-fluoro modifications; (vii) the dsRNA comprises a duplex region of 12-40 nucleotide pairs in length; and (viii) the dsRNA has a blunt end at 5′-end of the antisense strand. 
     In some embodiments, the antisense strand comprises phosphorothioate internucleotide linkages between nucleotide positions 1 and 2, between nucleotide positions 2 and 3, between nucleotide positions 21 and 22, and between nucleotide positions 22 and 23, wherein the antisense strand contains at least one thermally destabilizing modification of the duplex located in the seed region of the antisense strand (i.e., at position 2-9 of the 5′-end of the antisense strand), and wherein the dsRNA optionally further has at least one (e.g., one, two, three, four, five, six, seven or all eight) of the following characteristics: (i) the antisense comprises 2, 3, 4, 5, or 6 2′-fluoro modifications; (ii) the sense strand is conjugated with a ligand; (iii) the sense strand comprises 2, 3, 4 or 5 2′-fluoro modifications; (iv) the sense strand comprises 1, 2, 3, 4, or 5 phosphorothioate internucleotide linkages; (v) the dsRNA comprises at least four 2′-fluoro modifications; (vi) the dsRNA comprises a duplex region of 12-40 nucleotide pairs in length; (vii) the dsRNA comprises a duplex region of 12-40 nucleotide pairs in length; and (viii) the dsRNA has a blunt end at 5′-end of the antisense strand. 
     In some embodiments, the sense strand comprises phosphorothioate internucleotide linkages between nucleotide positions 1 and 2, and between nucleotide positions 2 and 3, wherein the antisense strand contains at least one thermally destabilizing modification of the duplex located in the seed region of the antisense strand (i.e., at position 2-9 of the 5′-end of the antisense strand), and wherein the dsRNA optionally further has at least one (e.g., one, two, three, four, five, six, seven or all eight) of the following characteristics: (i) the antisense comprises 2, 3, 4, 5, or 6 2′-fluoro modifications; (ii) the antisense comprises 1, 2, 3, 4, or 5 phosphorothioate internucleotide linkages; (iii) the sense strand is conjugated with a ligand; (iv) the sense strand comprises 2, 3, 4 or 5 2′-fluoro modifications; (v) the sense strand comprises 3, 4 or 5 phosphorothioate internucleotide linkages; (vi) the dsRNA comprises at least four 2′-fluoro modifications; (vii) the dsRNA comprises a duplex region of 12-40 nucleotide pairs in length; and (viii) the dsRNA has a blunt end at 5′-end of the antisense strand. 
     In some embodiments, the sense strand comprises phosphorothioate internucleotide linkages between nucleotide positions 1 and 2, and between nucleotide positions 2 and 3, the antisense strand comprises phosphorothioate internucleotide linkages between nucleotide positions 1 and 2, between nucleotide positions 2 and 3, between nucleotide positions 21 and 22, and between nucleotide positions 22 and 23, wherein the antisense strand contains at least one thermally destabilizing modification of the duplex located in the seed region of the antisense strand (i.e., at position 2-9 of the 5′-end of the antisense strand), and wherein the dsRNA optionally further has at least one (e.g., one, two, three, four, five, six or all seven) of the following characteristics: (i) the antisense comprises 2, 3, 4, 5 or 6 2′-fluoro modifications; (ii) the sense strand is conjugated with a ligand; (iii) the sense strand comprises 2, 3, 4 or 5 2′-fluoro modifications; (iv) the sense strand comprises 3, 4 or 5 phosphorothioate internucleotide linkages; (v) the dsRNA comprises at least four 2′-fluoro modifications; (vi) the dsRNA comprises a duplex region of 12-40 nucleotide pairs in length; and (vii) the dsRNA has a blunt end at 5′-end of the antisense strand. 
     In some embodiments, the dsRNA molecule of the disclosure comprises mismatch(es) with the target, within the duplex, or combinations thereof. The mismatch can occur in the overhang region or the duplex region. The base pair can be ranked on the basis of their propensity to promote dissociation or melting (e.g., on the free energy of association or dissociation of a particular pairing, the simplest approach is to examine the pairs on an individual pair basis, though next neighbor or similar analysis can also be used). In terms of promoting dissociation: A:U is preferred over G:C; G:U is preferred over G:C; and I:C is preferred over G:C (I=inosine). Mismatches, e.g., non-canonical or other than canonical pairings (as described elsewhere herein) are preferred over canonical (A:T, A:U, G:C) pairings; and pairings which include a universal base are preferred over canonical pairings. 
     In some embodiments, the dsRNA molecule of the disclosure comprises at least one of the first 1, 2, 3, 4, or 5 base pairs within the duplex regions from the 5′-end of the antisense strand can be chosen independently from the group of: A:U, G:U, I:C, and mismatched pairs, e.g., non-canonical or other than canonical pairings or pairings which include a universal base, to promote the dissociation of the antisense strand at the 5′-end of the duplex. 
     In some embodiments, the nucleotide at the 1 position within the duplex region from the 5′-end in the antisense strand is selected from the group consisting of A, dA, dU, U, and dT. Alternatively, at least one of the first 1, 2 or 3 base pair within the duplex region from the 5′-end of the antisense strand is an AU base pair. For example, the first base pair within the duplex region from the 5′-end of the antisense strand is an AU base pair. 
     It was found that introducing 4′-modified or 5′-modified nucleotide to the 3′-end of a phosphodiester (PO), phosphorothioate (PS), or phosphorodithioate (PS2) linkage of a dinucleotide at any position of single stranded or double stranded oligonucleotide can exert steric effect to the internucleotide linkage and, hence, protecting or stabilizing it against nucleases. 
     In some embodiments, 5′-modified nucleoside is introduced at the 3′-end of a dinucleotide at any position of single stranded or double stranded siRNA. For instance, a 5′-alkylated nucleoside may be introduced at the 3′-end of a dinucleotide at any position of single stranded or double stranded siRNA. The alkyl group at the 5′ position of the ribose sugar can be racemic or chirally pure R or S isomer. An exemplary 5′-alkylated nucleoside is 5′-methyl nucleoside. The 5′-methyl can be either racemic or chirally pure R or S isomer. 
     In some embodiments, 4′-modified nucleoside is introduced at the 3′-end of a dinucleotide at any position of single stranded or double stranded siRNA. For instance, a 4′-alkylated nucleoside may be introduced at the 3′-end of a dinucleotide at any position of single stranded or double stranded siRNA. The alkyl group at the 4′ position of the ribose sugar can be racemic or chirally pure R or S isomer. An exemplary 4′-alkylated nucleoside is 4′-methyl nucleoside. The 4′-methyl can be either racemic or chirally pure R or S isomer. Alternatively, a 4′-O-alkylated nucleoside may be introduced at the 3′-end of a dinucleotide at any position of single stranded or double stranded siRNA. The 4′-O-alkyl of the ribose sugar can be racemic or chirally pure R or S isomer. An exemplary 4′-O-alkylated nucleoside is 4′-O-methyl nucleoside. The 4′-O-methyl can be either racemic or chirally pure R or S isomer. 
     In some embodiments, 5′-alkylated nucleoside is introduced at any position on the sense strand or antisense strand of a dsRNA, and such modification maintains or improves potency of the dsRNA. The 5′-alkyl can be either racemic or chirally pure R or S isomer. An exemplary 5′-alkylated nucleoside is 5′-methyl nucleoside. The 5′-methyl can be either racemic or chirally pure R or S isomer. 
     In some embodiments, 4′-alkylated nucleoside is introduced at any position on the sense strand or antisense strand of a dsRNA, and such modification maintains or improves potency of the dsRNA. The 4′-alkyl can be either racemic or chirally pure R or S isomer. An exemplary 4′-alkylated nucleoside is 4′-methyl nucleoside. The 4′-methyl can be either racemic or chirally pure R or S isomer. 
     In some embodiments, 4′-O-alkylated nucleoside is introduced at any position on the sense strand or antisense strand of a dsRNA, and such modification maintains or improves potency of the dsRNA. The 5′-alkyl can be either racemic or chirally pure R or S isomer. An exemplary 4′-O-alkylated nucleoside is 4′-O-methyl nucleoside. The 4′-O-methyl can be either racemic or chirally pure R or S isomer. 
     In some embodiments, the dsRNA molecule of the disclosure can comprise 2′-5′ linkages (with 2′-H, 2′-OH and 2′-OMe and with P═O or P═S). For example, the 2′-5′ linkages modifications can be used to promote nuclease resistance or to inhibit binding of the sense to the antisense strand, or can be used at the 5′ end of the sense strand to avoid sense strand activation by RISC. 
     In another embodiment, the dsRNA molecule of the disclosure can comprise L sugars (e.g., L ribose, L-arabinose with 2′-H, 2′-OH and 2′-OMe). For example, these L sugars modifications can be used to promote nuclease resistance or to inhibit binding of the sense to the antisense strand, or can be used at the 5′ end of the sense strand to avoid sense strand activation by RISC. 
     Various publications describe multimeric siRNA which can all be used with the dsRNA of the disclosure. Such publications include WO2007/091269, U.S. Pat. No. 7,858,769, WO2010/141511, WO2007/117686, WO2009/014887, and WO2011/031520 which are hereby incorporated by their entirely. 
     As described in more detail below, the RNAi agent that contains conjugations of one or more carbohydrate moieties to an RNAi agent can optimize one or more properties of the RNAi agent. In many cases, the carbohydrate moiety will be attached to a modified subunit of the RNAi agent. For example, the ribose sugar of one or more ribonucleotide subunits of a dsRNA agent can be replaced with another moiety, e.g., a non-carbohydrate (preferably cyclic) carrier to which is attached a carbohydrate ligand. A ribonucleotide subunit in which the ribose sugar of the subunit has been so replaced is referred to herein as a ribose replacement modification subunit (RRMS). A cyclic carrier may be a carbocyclic ring system, i.e., all ring atoms are carbon atoms, or a heterocyclic ring system, i.e., one or more ring atoms may be a heteroatom, e.g., nitrogen, oxygen, sulfur. The cyclic carrier may be a monocyclic ring system, or may contain two or more rings, e.g. fused rings. The cyclic carrier may be a fully saturated ring system, or it may contain one or more double bonds. 
     The ligand may be attached to the polynucleotide via a carrier. The carriers include (i) at least one “backbone attachment point,” preferably two “backbone attachment points” and (ii) at least one “tethering attachment point.” A “backbone attachment point” as used herein refers to a functional group, e.g. a hydroxyl group, or generally, a bond available for, and that is suitable for incorporation of the carrier into the backbone, e.g., the phosphate, or modified phosphate, e.g., sulfur containing, backbone, of a ribonucleic acid. A “tethering attachment point” (TAP) in some embodiments refers to a constituent ring atom of the cyclic carrier, e.g., a carbon atom or a heteroatom (distinct from an atom which provides a backbone attachment point), that connects a selected moiety. The moiety can be, e.g., a carbohydrate, e.g. monosaccharide, disaccharide, trisaccharide, tetrasaccharide, oligosaccharide and polysaccharide. Optionally, the selected moiety is connected by an intervening tether to the cyclic carrier. Thus, the cyclic carrier will often include a functional group, e.g., an amino group, or generally, provide a bond, that is suitable for incorporation or tethering of another chemical entity, e.g., a ligand to the constituent ring. 
     The RNAi agents may be conjugated to a ligand via a carrier, wherein the carrier can be cyclic group or acyclic group; preferably, the cyclic group is selected from pyrrolidinyl, pyrazolinyl, pyrazolidinyl, imidazolinyl, imidazolidinyl, piperidinyl, piperazinyl, [1,3]dioxolane, oxazolidinyl, isoxazolidinyl, morpholinyl, thiazolidinyl, isothiazolidinyl, quinoxalinyl, pyridazinonyl, tetrahydrofuryl and and decalin; preferably, the acyclic group is selected from serinol backbone or diethanolamine backbone. 
     In certain specific embodiments, the RNAi agent for use in the methods of the disclosure is an agent selected from the group of agents listed in any one of Tables 2-5. These agents may further comprise a ligand, such as one or more lipophilic moieties, one or more GalNAc derivatives, or both of one of more lipophilic moieties and one or more GalNAc derivatives. 
     IV. IRNAS CONJUGATED TO LIGANDS 
     Another modification of the RNA of an iRNA of the invention involves chemically linking to the iRNA one or more ligands, moieties or conjugates that enhance the activity, cellular distribution or cellular uptake of the iRNA, e.g., into a cell. Such moieties include but are not limited to lipid moieties such as a cholesterol moiety (Letsinger et al.,  Proc. Natl. Acid. Sci. USA,  1989, 86: 6553-6556), cholic acid (Manoharan et al.,  Biorg. Med. Chem. Let.,  1994, 4:1053-1060), a thioether, e.g., beryl-S-tritylthiol (Manoharan et al.,  Ann. N.Y. Acad. Sci.,  1992, 660:306-309; Manoharan et al.,  Biorg. Med. Chem. Let.,  1993, 3:2765-2770), a thiocholesterol (Oberhauser et al.,  Nucl. Acids Res.,  1992, 20:533-538), an aliphatic chain, e.g., dodecandiol or undecyl residues (Saison-Behmoaras et al.,  EMBO J,  1991, 10:1111-1118; Kabanov et al.,  FEBS Lett.,  1990, 259:327-330; Svinarchuk et al.,  Biochimie,  1993, 75:49-54), a phospholipid, e.g., di-hexadecyl-rac-glycerol or triethyl-ammonium 1,2-di-O-hexadecyl-rac-glycero-3-phosphonate (Manoharan et al.,  Tetrahedron Lett.,  1995, 36:3651-3654; Shea et al.,  Nucl. Acids Res.,  1990, 18:3777-3783), a polyamine or a polyethylene glycol chain (Manoharan et al.,  Nucleosides  &amp;  Nucleotides,  1995, 14:969-973), or adamantane acetic acid (Manoharan et al.,  Tetrahedron Lett.,  1995, 36:3651-3654), a palmityl moiety (Mishra et al.,  Biochim. Biophys. Acta,  1995, 1264:229-237), or an octadecylamine or hexylamino-carbonyloxycholesterol moiety (Crooke et al.,  J. Pharmacol. Exp. Ther.,  1996, 277:923-937). 
     In certain embodiments, a ligand alters the distribution, targeting or lifetime of an iRNA agent into which it is incorporated. In some embodiments, a ligand provides an enhanced affinity for a selected target, e.g., molecule, cell or cell type, compartment, e.g., a cellular or organ compartment, tissue, organ or region of the body, as, e.g., compared to a species absent such a ligand. Typical ligands will not take part in duplex pairing in a duplexed nucleic acid. 
     Ligands can include a naturally occurring substance, such as a protein (e.g., human serum albumin (HSA), low-density lipoprotein (LDL), or globulin); carbohydrate (e.g., a dextran, pullulan, chitin, chitosan, inulin, cyclodextrin or hyaluronic acid); or a lipid. The ligand may also be a recombinant or synthetic molecule, such as a synthetic polymer, e.g., a synthetic polyamino acid. Examples of polyamino acids include polyamino acid is a polylysine (PLL), poly L-aspartic acid, poly L-glutamic acid, styrene-maleic acid anhydride copolymer, poly(L-lactide-co-glycolied) copolymer, divinyl ether-maleic anhydride copolymer, N-(2-hydroxypropyl)methacrylamide copolymer (HMPA), polyethylene glycol (PEG), polyvinyl alcohol (PVA), polyurethane, poly(2-ethylacryllic acid), N-isopropylacrylamide polymers, or polyphosphazine. Example of polyamines include: polyethylenimine, polylysine (PLL), spermine, spermidine, polyamine, pseudopeptide-polyamine, peptidomimetic polyamine, dendrimer polyamine, arginine, amidine, protamine, cationic lipid, cationic porphyrin, quaternary salt of a polyamine, or an α helical peptide. 
     Ligands can also include targeting groups, e.g., a cell or tissue targeting agent, e.g., a lectin, glycoprotein, lipid or protein, e.g., an antibody, that binds to a specified cell type such as a kidney cell. A targeting group can be a thyrotropin, melanotropin, lectin, glycoprotein, surfactant protein A, Mucin carbohydrate, multivalent lactose, multivalent galactose, N-acetyl-galactosamine, N-acetyl-glucosamine multivalent mannose, multivalent fucose, glycosylated polyaminoacids, multivalent galactose, transferrin, bisphosphonate, polyglutamate, polyaspartate, a lipid, cholesterol, a steroid, bile acid, folate, vitamin B12, biotin, or an RGD peptide or RGD peptide mimetic. In certain embodiments, the ligand is a multivalent galactose, e.g., an N-acetyl-galactosamine 
     Other examples of ligands include dyes, intercalating agents (e.g. acridines), cross-linkers (e.g. psoralene, mitomycin C), porphyrins (TPPC4, texaphyrin, Sapphyrin), polycyclic aromatic hydrocarbons (e.g., phenazine, dihydrophenazine), artificial endonucleases (e.g. EDTA), lipophilic molecules, e.g., cholesterol, cholic acid, adamantane acetic acid, 1-pyrene butyric acid, dihydrotestosterone, 1,3-Bis-O(hexadecyl)glycerol, geranyloxyhexyl group, hexadecylglycerol, borneol, menthol, 1,3-propanediol, heptadecyl group, palmitic acid, myristic acid, O3-(oleoyl)lithocholic acid, O3-(oleoyl)cholenic acid, dimethoxytrityl, or phenoxazine) and peptide conjugates (e.g., antennapedia peptide, Tat peptide), alkylating agents, phosphate, amino, mercapto, PEG (e.g., PEG-40K), MPEG, [MPEG]2, polyamino, alkyl, substituted alkyl, radiolabeled markers, enzymes, haptens (e.g. biotin), transport/absorption facilitators (e.g., aspirin, vitamin E, folic acid), synthetic ribonucleases (e.g., imidazole, bisimidazole, histamine, imidazole clusters, acridine-imidazole conjugates, Eu3+ complexes of tetraazamacrocycles), dinitrophenyl, HRP, or AP. 
     Ligands can be proteins, e.g., glycoproteins, or peptides, e.g., molecules having a specific affinity for a co-ligand, or antibodies e.g., an antibody, that binds to a specified cell type such as a cancer cell, endothelial cell, or bone cell. Ligands may also include hormones and hormone receptors. They can also include non-peptidic species, such as lipids, lectins, carbohydrates, vitamins, cofactors, multivalent lactose, multivalent galactose, N-acetyl-galactosamine, N-acetyl-glucosamine multivalent mannose, or multivalent fucose. The ligand can be, for example, a lipopolysaccharide, an activator of p38 MAP kinase, or an activator of NF-κB. 
     The ligand can be a substance, e.g., a drug, which can increase the uptake of the iRNA agent into the cell, for example, by disrupting the cell&#39;s cytoskeleton, e.g., by disrupting the cell&#39;s microtubules, microfilaments, or intermediate filaments. The drug can be, for example, taxon, vincristine, vinblastine, cytochalasin, nocodazole, japlakinolide, latrunculin A, phalloidin, swinholide A, indanocine, or myoservin. 
     In some embodiments, a ligand attached to an iRNA as described herein acts as a pharmacokinetic modulator (PK modulator). PK modulators include lipophiles, bile acids, steroids, phospholipid analogues, peptides, protein binding agents, PEG, vitamins etc. Exemplary PK modulators include, but are not limited to, cholesterol, fatty acids, cholic acid, lithocholic acid, dialkylglycerides, diacylglyceride, phospholipids, sphingolipids, naproxen, ibuprofen, vitamin E, biotin etc. Oligonucleotides that comprise a number of phosphorothioate linkages are also known to bind to serum protein, thus short oligonucleotides, e.g., oligonucleotides of about 5 bases, 10 bases, 15 bases or 20 bases, comprising multiple of phosphorothioate linkages in the backbone are also amenable to the present invention as ligands (e.g. as PK modulating ligands). In addition, aptamers that bind serum components (e.g. serum proteins) are also suitable for use as PK modulating ligands in the embodiments described herein. 
     Ligand-conjugated iRNAs of the invention may be synthesized by the use of an oligonucleotide that bears a pendant reactive functionality, such as that derived from the attachment of a linking molecule onto the oligonucleotide (described below). This reactive oligonucleotide may be reacted directly with commercially-available ligands, ligands that are synthesized bearing any of a variety of protecting groups, or ligands that have a linking moiety attached thereto. 
     The oligonucleotides used in the conjugates of the present invention may be conveniently and routinely made through the well-known technique of solid-phase synthesis. Equipment for such synthesis is sold by several vendors including, for example, Applied Biosystems® (Foster City, Calif.). Any other means for such synthesis known in the art may additionally or alternatively be employed. It is also known to use similar techniques to prepare other oligonucleotides, such as the phosphorothioates and alkylated derivatives. 
     In the ligand-conjugated oligonucleotides and ligand-molecule bearing sequence-specific linked nucleosides of the present invention, the oligonucleotides and oligonucleosides may be assembled on a suitable DNA synthesizer utilizing standard nucleotide or nucleoside precursors, or nucleotide or nucleoside conjugate precursors that already bear the linking moiety, ligand-nucleotide or nucleoside-conjugate precursors that already bear the ligand molecule, or non-nucleoside ligand-bearing building blocks. 
     When using nucleotide-conjugate precursors that already bear a linking moiety, the synthesis of the sequence-specific linked nucleosides is typically completed, and the ligand molecule is then reacted with the linking moiety to form the ligand-conjugated oligonucleotide. In some embodiments, the oligonucleotides or linked nucleosides of the present invention are synthesized by an automated synthesizer using phosphoramidites derived from ligand-nucleoside conjugates in addition to the standard phosphoramidites and non-standard phosphoramidites that are commercially available and routinely used in oligonucleotide synthesis. 
     A. Lipid Conjugates 
     In certain embodiments, the ligand or conjugate is a lipid or lipid-based molecule. Such a lipid or lipid-based molecule can typically bind a serum protein, such as human serum albumin (HSA). An HSA binding ligand allows for distribution of the conjugate to a target tissue, e.g., a non-kidney target tissue of the body. For example, the target tissue can be the liver, including parenchymal cells of the liver. Other molecules that can bind HSA can also be used as ligands. For example, naproxen or aspirin can be used. A lipid or lipid-based ligand can (a) increase resistance to degradation of the conjugate, (b) increase targeting or transport into a target cell or cell membrane, or (c) can be used to adjust binding to a serum protein, e.g., HSA. 
     A lipid-based ligand can be used to modulate, e.g., control (e.g., inhibit) the binding of the conjugate to a target tissue. For example, a lipid or lipid-based ligand that binds to HSA more strongly will be less likely to be targeted to the kidney and therefore less likely to be cleared from the body. A lipid or lipid-based ligand that binds to HSA less strongly can be used to target the conjugate to the kidney. 
     In certain embodiments, the lipid-based ligand binds HSA. For example, the ligand can bind HSA with a sufficient affinity such that distribution of the conjugate to a non-kidney tissue is enhanced. However, the affinity is typically not so strong that the HSA-ligand binding cannot be reversed. 
     In certain embodiments, the lipid-based ligand binds HSA weakly or not at all, such that distribution of the conjugate to the kidney is enhanced. Other moieties that target to kidney cells can also be used in place of or in addition to the lipid-based ligand. 
     In another aspect, the ligand is a moiety, e.g., a vitamin, which is taken up by a target cell, e.g., a proliferating cell. These are particularly useful for treating disorders characterized by unwanted cell proliferation, e.g., of the malignant or non-malignant type, e.g., cancer cells. Exemplary vitamins include vitamin A, E, and K. Other exemplary vitamins include are B vitamin, e.g., folic acid, B12, riboflavin, biotin, pyridoxal or other vitamins or nutrients taken up by cancer cells. Also included are HSA and low density lipoprotein (LDL). 
     B. Cell Permeation Agents 
     In another aspect, the ligand is a cell-permeation agent, such as a helical cell-permeation agent. In certain embodiments, the agent is amphipathic. An exemplary agent is a peptide such as tat or antennopedia. If the agent is a peptide, it can be modified, including a peptidylmimetic, invertomers, non-peptide or pseudo-peptide linkages, and use of D-amino acids. The helical agent is typically an α-helical agent and can have a lipophilic and a lipophobic phase. 
     The ligand can be a peptide or peptidomimetic. A peptidomimetic (also referred to herein as an oligopeptidomimetic) is a molecule capable of folding into a defined three-dimensional structure similar to a natural peptide. The attachment of peptide and peptidomimetics to iRNA agents can affect pharmacokinetic distribution of the iRNA, such as by enhancing cellular recognition and absorption. The peptide or peptidomimetic moiety can be about 5-50 amino acids long, e.g., about 5, 10, 15, 20, 25, 30, 35, 40, 45, or 50 amino acids long. 
     A peptide or peptidomimetic can be, for example, a cell permeation peptide, cationic peptide, amphipathic peptide, or hydrophobic peptide (e.g., consisting primarily of Tyr, Trp, or Phe). The peptide moiety can be a dendrimer peptide, constrained peptide or crosslinked peptide. In another alternative, the peptide moiety can include a hydrophobic membrane translocation sequence (MTS). An exemplary hydrophobic MTS-containing peptide is RFGF having the amino acid sequence AAVALLPAVLLALLAP (SEQ ID NO:13). An RFGF analogue (e.g., amino acid sequence AALLPVLLAAP (SEQ ID NO:14)) containing a hydrophobic MTS can also be a targeting moiety. The peptide moiety can be a “delivery” peptide, which can carry large polar molecules including peptides, oligonucleotides, and protein across cell membranes. For example, sequences from the HIV Tat protein (GRKKRRQRRRPPQ (SEQ ID NO: 15)) and the  Drosophila  Antennapedia protein (RQIKIWFQNRRMKWKK (SEQ ID NO:16)) have been found to be capable of functioning as delivery peptides. A peptide or peptidomimetic can be encoded by a random sequence of DNA, such as a peptide identified from a phage-display library, or one-bead-one-compound (OBOC) combinatorial library (Lam et al.,  Nature,  354:82-84, 1991). Typically, the peptide or peptidomimetic tethered to a dsRNA agent via an incorporated monomer unit is a cell targeting peptide such as an arginine-glycine-aspartic acid (RGD)-peptide, or RGD mimic A peptide moiety can range in length from about 5 amino acids to about 40 amino acids. The peptide moieties can have a structural modification, such as to increase stability or direct conformational properties. Any of the structural modifications described below can be utilized. 
     An RGD peptide for use in the compositions and methods of the invention may be linear or cyclic, and may be modified, e.g., glycosylated or methylated, to facilitate targeting to a specific tissue(s). RGD-containing peptides and peptidiomimemtics may include D-amino acids, as well as synthetic RGD mimics. In addition to RGD, one can use other moieties that target the integrin ligand. Preferred conjugates of this ligand target PECAM-1 or VEGF. 
     An RGD peptide moiety can be used to target a particular cell type, e.g., a tumor cell, such as an endothelial tumor cell or a breast cancer tumor cell (Zitzmann et al.,  Cancer Res.,  62:5139-43, 2002). An RGD peptide can facilitate targeting of an dsRNA agent to tumors of a variety of other tissues, including the lung, kidney, spleen, or liver (Aoki et al.,  Cancer Gene Therapy  8:783-787, 2001). Typically, the RGD peptide will facilitate targeting of an iRNA agent to the kidney. The RGD peptide can be linear or cyclic, and can be modified, e.g., glycosylated or methylated to facilitate targeting to specific tissues. For example, a glycosylated RGD peptide can deliver an iRNA agent to a tumor cell expressing α v β 3  (Haubner et al.,  Jour. Nucl. Med.,  42:326-336, 2001). 
     A “cell permeation peptide” is capable of permeating a cell, e.g., a microbial cell, such as a bacterial or fungal cell, or a mammalian cell, such as a human cell. A microbial cell-permeating peptide can be, for example, an α-helical linear peptide (e.g., LL-37 or Ceropin P1), a disulfide bond-containing peptide (e.g., α-defensin, β-defensin or bactenecin), or a peptide containing only one or two dominating amino acids (e.g., PR-39 or indolicidin). A cell permeation peptide can also include a nuclear localization signal (NLS). For example, a cell permeation peptide can be a bipartite amphipathic peptide, such as MPG, which is derived from the fusion peptide domain of HIV-1 gp41 and the NLS of SV40 large T antigen (Simeoni et al.,  Nucl. Acids Res.  31:2717-2724, 2003). 
     C. Carbohydrate Conjugates 
     In some embodiments of the compositions and methods of the invention, an iRNA further comprises a carbohydrate. The carbohydrate conjugated iRNA are advantageous for the in vivo delivery of nucleic acids, as well as compositions suitable for in vivo therapeutic use, as described herein. As used herein, “carbohydrate” refers to a compound which is either a carbohydrate per se made up of one or more monosaccharide units having at least 6 carbon atoms (which can be linear, branched or cyclic) with an oxygen, nitrogen or sulfur atom bonded to each carbon atom; or a compound having as a part thereof a carbohydrate moiety made up of one or more monosaccharide units each having at least six carbon atoms (which can be linear, branched or cyclic), with an oxygen, nitrogen or sulfur atom bonded to each carbon atom. Representative carbohydrates include the sugars (mono-, di-, tri- and oligosaccharides containing from about 4, 5, 6, 7, 8, or 9 monosaccharide units), and polysaccharides such as starches, glycogen, cellulose and polysaccharide gums. Specific monosaccharides include C5 and above (e.g., C5, C6, C7, or C8) sugars; di- and tri-saccharides include sugars having two or three monosaccharide units (e.g., C5, C6, C7, or C8). 
     In certain embodiments, a carbohydrate conjugate comprises a monosaccharide. 
     In certain embodiments, the monosaccharide is an N-acetylgalactosamine (GalNAc). GalNAc conjugates, which comprise one or more N-acetylgalactosamine (GalNAc) derivatives, are described, for example, in U.S. Pat. No. 8,106,022, the entire content of which is hereby incorporated herein by reference. In some embodiments, the GalNAc conjugate serves as a ligand that targets the iRNA to particular cells. In some embodiments, the GalNAc conjugate targets the iRNA to liver cells, e.g., by serving as a ligand for the asialoglycoprotein receptor of liver cells (e.g., hepatocytes). 
     In some embodiments, the carbohydrate conjugate comprises one or more GalNAc derivatives. The GalNAc derivatives may be attached via a linker, e.g., a bivalent or trivalent branched linker. In some embodiments the GalNAc conjugate is conjugated to the 3′ end of the sense strand. In some embodiments, the GalNAc conjugate is conjugated to the iRNA agent (e.g., to the 3′ end of the sense strand) via a linker, e.g., a linker as described herein. In some embodiments the GalNAc conjugate is conjugated to the 5′ end of the sense strand. In some embodiments, the GalNAc conjugate is conjugated to the iRNA agent (e.g., to the 5′ end of the sense strand) via a linker, e.g., a linker as described herein. 
     In certain embodiments of the invention, the GalNAc or GalNAc derivative is attached to an iRNA agent of the invention via a monovalent linker. In some embodiments, the GalNAc or GalNAc derivative is attached to an iRNA agent of the invention via a bivalent linker. In yet other embodiments of the invention, the GalNAc or GalNAc derivative is attached to an iRNA agent of the invention via a trivalent linker. In other embodiments of the invention, the GalNAc or GalNAc derivative is attached to an iRNA agent of the invention via a tetravalent linker. 
     In certain embodiments, the double stranded RNAi agents of the invention comprise one GalNAc or GalNAc derivative attached to the iRNA agent. In certain embodiments, the double stranded RNAi agents of the invention comprise a plurality (e.g., 2, 3, 4, 5, or 6) GalNAc or GalNAc derivatives, each independently attached to a plurality of nucleotides of the double stranded RNAi agent through a plurality of monovalent linkers. 
     In some embodiments, for example, when the two strands of an iRNA agent of the invention are part of one larger molecule connected by an uninterrupted chain of nucleotides between the 3′-end of one strand and the 5′-end of the respective other strand forming a hairpin loop comprising, a plurality of unpaired nucleotides, each unpaired nucleotide within the hairpin loop may independently comprise a GalNAc or GalNAc derivative attached via a monovalent linker. The hairpin loop may also be formed by an extended overhang in one strand of the duplex. 
     In some embodiments, for example, when the two strands of an iRNA agent of the invention are part of one larger molecule connected by an uninterrupted chain of nucleotides between the 3′-end of one strand and the 5′-end of the respective other strand forming a hairpin loop comprising, a plurality of unpaired nucleotides, each unpaired nucleotide within the hairpin loop may independently comprise a GalNAc or GalNAc derivative attached via a monovalent linker. The hairpin loop may also be formed by an extended overhang in one strand of the duplex. 
     In some embodiments, the GalNAc conjugate is 
     
       
         
         
             
             
         
       
     
     In some embodiments, the RNAi agent is attached to the carbohydrate conjugate via a linker as shown in the following schematic, wherein X is O or S 
     
       
         
         
             
             
         
       
     
     In some embodiments, the RNAi agent is conjugated to L96 as defined in Table 1 and shown below: 
     
       
         
         
             
             
         
       
     
     In certain embodiments, a carbohydrate conjugate for use in the compositions and methods of the invention is selected from the group consisting of: 
     
       
         
         
             
             
         
       
       
         
         
             
             
         
       
       
         
         
             
             
         
       
       
         
         
             
             
         
       
       
         
         
             
             
         
       
       
         
         
             
             
         
       
       
         
         
             
             
         
       
       
         
         
             
             
         
       
       
         
         
             
             
         
       
     
     In certain embodiments, a carbohydrate conjugate for use in the compositions and methods of the invention is a monosaccharide. In certain embodiments, the monosaccharide is an N-acetylgalactosamine, such as 
     
       
         
         
             
             
         
       
     
     Another representative carbohydrate conjugate for use in the embodiments described herein includes, but is not limited to, 
     
       
         
         
             
             
         
       
     
     when one of X or Y is an oligonucleotide, the other is a hydrogen. 
     In some embodiments, a suitable ligand is a ligand disclosed in WO 2019/055633, the entire contents of which are incorporated herein by reference. In one embodiment the ligand comprises the structure below: 
     
       
         
         
             
             
         
       
     
     In certain embodiments, the RNAi agents of the disclosure may include GalNAc ligands, even if such GalNAc ligands are currently projected to be of limited value for the preferred pulmonary system delivery route(s) of the instant disclosure. 
     In certain embodiments of the invention, the GalNAc or GalNAc derivative is attached to an iRNA agent of the invention via a monovalent linker. In some embodiments, the GalNAc or GalNAc derivative is attached to an iRNA agent of the invention via a bivalent linker. In yet other embodiments of the invention, the GalNAc or GalNAc derivative is attached to an iRNA agent of the invention via a trivalent linker. In other embodiments of the invention, the GalNAc or GalNAc derivative is attached to an iRNA agent of the invention via a tetravalent linker. 
     In certain embodiments, the double stranded RNAi agents of the invention comprise one GalNAc or GalNAc derivative attached to the iRNA agent, e.g., the 5′ end of the sense strand of a dsRNA agent, or the 5′ end of one or both sense strands of a dual targeting RNAi agent as described herein. In certain embodiments, the double stranded RNAi agents of the invention comprise a plurality (e.g., 2, 3, 4, 5, or 6) GalNAc or GalNAc derivatives, each independently attached to a plurality of nucleotides of the double stranded RNAi agent through a plurality of monovalent linkers. 
     In some embodiments, for example, when the two strands of an iRNA agent of the invention are part of one larger molecule connected by an uninterrupted chain of nucleotides between the 3′-end of one strand and the 5′-end of the respective other strand forming a hairpin loop comprising, a plurality of unpaired nucleotides, each unpaired nucleotide within the hairpin loop may independently comprise a GalNAc or GalNAc derivative attached via a monovalent linker. 
     In some embodiments, the carbohydrate conjugate further comprises one or more additional ligands as described above, such as, but not limited to, a PK modulator or a cell permeation peptide. Additional carbohydrate conjugates and linkers suitable for use in the present invention include those described in WO 2014/179620 and WO 2014/179627, the entire contents of each of which are incorporated herein by reference. 
     D. Linkers 
     In some embodiments, the conjugate or ligand described herein can be attached to an iRNA oligonucleotide with various linkers that can be cleavable or non-cleavable. 
     The term “linker” or “linking group” means an organic moiety that connects two parts of a compound, e.g., covalently attaches two parts of a compound. Linkers typically comprise a direct bond or an atom such as oxygen or sulfur, a unit such as NR8, C(O), C(O)NH, SO, SO 2 , SO 2 NH or a chain of atoms, such as, but not limited to, substituted or unsubstituted alkyl, substituted or unsubstituted alkenyl, substituted or unsubstituted alkynyl, arylalkyl, arylalkenyl, arylalkynyl, heteroarylalkyl, heteroarylalkenyl, heteroarylalkynyl, heterocyclylalkyl, heterocyclylalkenyl, heterocyclylalkynyl, aryl, heteroaryl, heterocyclyl, cycloalkyl, cycloalkenyl, alkylarylalkyl, alkylarylalkenyl, alkylarylalkynyl, alkenylarylalkyl, alkenylarylalkenyl, alkenylarylalkynyl, alkynylarylalkyl, alkynylarylalkenyl, alkynylarylalkynyl, alkylheteroarylalkyl, alkylheteroarylalkenyl, alkylheteroarylalkynyl, alkenylheteroarylalkyl, alkenylheteroarylalkenyl, alkenylheteroarylalkynyl, alkynylheteroarylalkyl, alkynylheteroarylalkenyl, alkynylheteroarylalkynyl, alkylheterocyclylalkyl, alkylheterocyclylalkenyl, alkylhererocyclylalkynyl, alkenylheterocyclylalkyl, alkenylheterocyclylalkenyl, alkenylheterocyclylalkynyl, alkynylheterocyclylalkyl, alkynylheterocyclylalkenyl, alkynylheterocyclylalkynyl, alkylaryl, alkenylaryl, alkynylaryl, alkylheteroaryl, alkenylheteroaryl, alkynylhereroaryl, which one or more methylenes can be interrupted or terminated by O, S, S(O), SO 2 , N(R8), C(O), substituted or unsubstituted aryl, substituted or unsubstituted heteroaryl, substituted or unsubstituted heterocyclic; where R8 is hydrogen, acyl, aliphatic or substituted aliphatic. In certain embodiments, the linker is between about 1-24 atoms, 2-24, 3-24, 4-24, 5-24, 6-24, 6-18, 7-18, 8-18 atoms, 7-17, 8-17, 6-16, 7-16, or 8-16 atoms. 
     A cleavable linking group is one which is sufficiently stable outside the cell, but which upon entry into a target cell is cleaved to release the two parts the linker is holding together. In a preferred embodiment, the cleavable linking group is cleaved at least about 10 times, 20, times, 30 times, 40 times, 50 times, 60 times, 70 times, 80 times, 90 times or more, or at least about 100 times faster in a target cell or under a first reference condition (which can, e.g., be selected to mimic or represent intracellular conditions) than in the blood of a subject, or under a second reference condition (which can, e.g., be selected to mimic or represent conditions found in the blood or serum). 
     Cleavable linking groups are susceptible to cleavage agents, e.g., pH, redox potential or the presence of degradative molecules. Generally, cleavage agents are more prevalent or found at higher levels or activities inside cells than in serum or blood. Examples of such degradative agents include: redox agents which are selected for particular substrates or which have no substrate specificity, including, e.g., oxidative or reductive enzymes or reductive agents such as mercaptans, present in cells, that can degrade a redox cleavable linking group by reduction; esterases; endosomes or agents that can create an acidic environment, e.g., those that result in a pH of five or lower; enzymes that can hydrolyze or degrade an acid cleavable linking group by acting as a general acid, peptidases (which can be substrate specific), and phosphatases. 
     A cleavable linkage group, such as a disulfide bond can be susceptible to pH. The pH of human serum is 7.4, while the average intracellular pH is slightly lower, ranging from about 7.1-7.3. Endosomes have a more acidic pH, in the range of 5.5-6.0, and lysosomes have an even more acidic pH at around 5.0. Some linkers will have a cleavable linking group that is cleaved at a preferred pH, thereby releasing a cationic lipid from the ligand inside the cell, or into the desired compartment of the cell. 
     A linker can include a cleavable linking group that is cleavable by a particular enzyme. The type of cleavable linking group incorporated into a linker can depend on the cell to be targeted. For example, a liver-targeting ligand can be linked to a cationic lipid through a linker that includes an ester group. Liver cells are rich in esterases, and therefore the linker will be cleaved more efficiently in liver cells than in cell types that are not esterase-rich. Other cell-types rich in esterases include cells of the lung, renal cortex, and testis. 
     Linkers that contain peptide bonds can be used when targeting cell types rich in peptidases, such as liver cells and synoviocytes. 
     In general, the suitability of a candidate cleavable linking group can be evaluated by testing the ability of a degradative agent (or condition) to cleave the candidate linking group. It will also be desirable to also test the candidate cleavable linking group for the ability to resist cleavage in the blood or when in contact with other non-target tissue. Thus, one can determine the relative susceptibility to cleavage between a first and a second condition, where the first is selected to be indicative of cleavage in a target cell and the second is selected to be indicative of cleavage in other tissues or biological fluids, e.g., blood or serum. The evaluations can be carried out in cell free systems, in cells, in cell culture, in organ or tissue culture, or in whole animals. It can be useful to make initial evaluations in cell-free or culture conditions and to confirm by further evaluations in whole animals. In preferred embodiments, useful candidate compounds are cleaved at least about 2, 4, 10, 20, 30, 40, 50, 60, 70, 80, 90, or about 100 times faster in the cell (or under in vitro conditions selected to mimic intracellular conditions) as compared to blood or serum (or under in vitro conditions selected to mimic extracellular conditions). 
     i. Redox Cleavable Linking Groups 
     In certain embodiments, a cleavable linking group is a redox cleavable linking group that is cleaved upon reduction or oxidation. An example of reductively cleavable linking group is a disulphide linking group (—S—S—). To determine if a candidate cleavable linking group is a suitable “reductively cleavable linking group,” or for example is suitable for use with a particular iRNA moiety and particular targeting agent one can look to methods described herein. For example, a candidate can be evaluated by incubation with dithiothreitol (DTT), or other reducing agent using reagents know in the art, which mimic the rate of cleavage which would be observed in a cell, e.g., a target cell. The candidates can also be evaluated under conditions which are selected to mimic blood or serum conditions. In one, candidate compounds are cleaved by at most about 10% in the blood. In other embodiments, useful candidate compounds are degraded at least about 2, 4, 10, 20, 30, 40, 50, 60, 70, 80, 90, or about 100 times faster in the cell (or under in vitro conditions selected to mimic intracellular conditions) as compared to blood (or under in vitro conditions selected to mimic extracellular conditions). The rate of cleavage of candidate compounds can be determined using standard enzyme kinetics assays under conditions chosen to mimic intracellular media and compared to conditions chosen to mimic extracellular media. 
     ii. Phosphate-Based Cleavable Linking Groups 
     In certain embodiments, a cleavable linker comprises a phosphate-based cleavable linking group. A phosphate-based cleavable linking group is cleaved by agents that degrade or hydrolyze the phosphate group. An example of an agent that cleaves phosphate groups in cells are enzymes such as phosphatases in cells. Examples of phosphate-based linking groups are —O—P(O)(ORk)-O—, —O—P(S)(ORk)-O—, —O—P(S)(SRk)-O—, —S—P(O)(ORk)-O—, —O—P(O)(ORk)-S—, —S—P(O)(ORk)-S—, —O—P(S)(ORk)-S—, —S—P(S)(ORk)-O—, —O—P(O)(Rk)-O—, —O—P(S)(Rk)-O—, —S—P(O)(Rk)-O—, —S—P(S)(Rk)-S—P(O)(Rk)-S—, —O—P(S)(Rk)-S. Preferred embodiments are —O—P(O)(OH)—O—, —O—P(S)(OH)—O—, P(S)(SH)—O—, —S—P(O)(OH)—O—, —O—P(O)(OH)—S—, —S—P(O)(OH)—S—, —O—P(S)(OH)—S—, —S—P(S)(OH)—O—P(O)(H)—O—, —O—P(S)(H)—O—, —S—P(O)(H)—O, —S—P(S)(H)—O—, —S—P(O)(H)—S—, —O—P(S)(H)—S—. A preferred embodiment is —O—P(O)(OH)—O—. These candidates can be evaluated using methods analogous to those described above. 
     iii. Acid Cleavable Linking Groups 
     In certain embodiments, a cleavable linker comprises an acid cleavable linking group. An acid cleavable linking group is a linking group that is cleaved under acidic conditions. In preferred embodiments acid cleavable linking groups are cleaved in an acidic environment with a pH of about 6.5 or lower (e.g., about 6.0, 5.75, 5.5, 5.25, 5.0, or lower), or by agents such as enzymes that can act as a general acid. In a cell, specific low pH organelles, such as endosomes and lysosomes can provide a cleaving environment for acid cleavable linking groups. Examples of acid cleavable linking groups include but are not limited to hydrazones, esters, and esters of amino acids. Acid cleavable groups can have the general formula —C═NN—, C(O)O, or —OC(O). A preferred embodiment is when the carbon attached to the oxygen of the ester (the alkoxy group) is an aryl group, substituted alkyl group, or tertiary alkyl group such as dimethyl pentyl or t-butyl. These candidates can be evaluated using methods analogous to those described above. 
     iv. Ester-Based Cleavable Linking Groups 
     In certain embodiments, a cleavable linker comprises an ester-based cleavable linking group. An ester-based cleavable linking group is cleaved by enzymes such as esterases and amidases in cells. Examples of ester-based cleavable linking groups include but are not limited to esters of alkylene, alkenylene and alkynylene groups. Ester cleavable linking groups have the general formula —C(O)O—, or —OC(O)—. These candidates can be evaluated using methods analogous to those described above. 
     v. Peptide-Based Cleavable Linking Groups 
     In yet another embodiment, a cleavable linker comprises a peptide-based cleavable linking group. A peptide-based cleavable linking group is cleaved by enzymes such as peptidases and proteases in cells. Peptide-based cleavable linking groups are peptide bonds formed between amino acids to yield oligopeptides (e.g., dipeptides, tripeptides etc.) and polypeptides. Peptide-based cleavable groups do not include the amide group (—C(O)NH—). The amide group can be formed between any alkylene, alkenylene or alkynelene. A peptide bond is a special type of amide bond formed between amino acids to yield peptides and proteins. The peptide based cleavage group is generally limited to the peptide bond (i.e., the amide bond) formed between amino acids yielding peptides and proteins and does not include the entire amide functional group. Peptide-based cleavable linking groups have the general formula —NHCHRAC(O)NHCHRBC(O)—, where RA and RB are the R groups of the two adjacent amino acids. These candidates can be evaluated using methods analogous to those described above. 
     In some embodiments, an iRNA of the invention is conjugated to a carbohydrate through a linker. Non-limiting examples of iRNA carbohydrate conjugates with linkers of the compositions and methods of the invention include, but are not limited to, 
     
       
         
         
             
             
         
       
       
         
         
             
             
         
       
       
         
         
             
             
         
       
     
     when one of X or Y is an oligonucleotide, the other is a hydrogen. 
     In certain embodiments of the compositions and methods of the invention, a ligand is one or more “GalNAc” (N-acetylgalactosamine) derivatives attached through a bivalent or trivalent branched linker. 
     In certain embodiments, a dsRNA of the invention is conjugated to a bivalent or trivalent branched linker selected from the group of structures shown in any of formula (XLV)-(XLVI): 
     
       
         
         
             
             
         
       
     
     wherein: 
     q2A, q2B, q3A, q3B, q4A, q4B, q5A, q5B and q5C represent independently for each occurrence 0-20 and wherein the repeating unit can be the same or different; 
     P 2A , P 2B , P 3A , P 3B , P 4A , P 4B , P 5A , P 5B , P 5C , T 2A , T 2B , T 3A , T 3B , T 4A , T 4B , T 4A , T 5B , T 5C  are each independently for each occurrence absent, CO, NH, O, S, OC(O), NHC(O), CH 2 , CH 2 NH or CH 2 O; 
     Q 2A , Q 2B , Q 3A , Q 3B , Q 4A , Q 4B , Q 5A , Q 5B , Q 5C  are independently for each occurrence absent, alkylene, substituted alkylene wherein one or more methylenes can be interrupted or terminated by one or more of O, S, S(O), SO 2 , N(R N ), C(R′)═C(R″), C≡C or C(O); 
     R 2A , R 2B , R 3A , R 3B , R 4A , R 4B , R 5A , R 5B , R 5C  are each independently for each occurrence absent, NH, O, S, CH 2 , C(O)O, C(O)NH, NHCH(R a )C(O), —C(O)—CH(R a )—NH—, CO, CH═N—O, 
     
       
         
         
             
             
         
       
     
     or heterocyclyl; 
     L 2A , L 2B , L 3A , L 3B , L 4A , L 4B , L 5A , L 5B , L 5C  represent the ligand; i.e. each independently for each occurrence a monosaccharide (such as GalNAc), disaccharide, trisaccharide, tetrasaccharide, oligosaccharide, or polysaccharide; and R a  is H or amino acid side chain. Trivalent conjugating GalNAc derivatives are particularly useful for use with RNAi agents for inhibiting the expression of a target gene, such as those of formula (XLIX): 
     
       
         
         
             
             
         
       
     
     wherein L 5A , L 5B  and L 5C  represent a monosaccharide, such as GalNAc derivative. 
     Examples of suitable bivalent and trivalent branched linker groups conjugating GalNAc derivatives include, but are not limited to, the structures recited above as formulas II, VII, XI, X, and XIII. 
     Representative U.S. patents that teach the preparation of RNA conjugates include, but are not limited to, U.S. Pat. Nos. 4,828,979; 4,948,882; 5,218,105; 5,525,465; 5,541,313; 5,545,730; 5,552,538; 5,578,717, 5,580,731; 5,591,584; 5,109,124; 5,118,802; 5,138,045; 5,414,077; 5,486,603; 5,512,439; 5,578,718; 5,608,046; 4,587,044; 4,605,735; 4,667,025; 4,762,779; 4,789,737; 4,824,941; 4,835,263; 4,876,335; 4,904,582; 4,958,013; 5,082,830; 5,112,963; 5,214,136; 5,082,830; 5,112,963; 5,214,136; 5,245,022; 5,254,469; 5,258,506; 5,262,536; 5,272,250; 5,292,873; 5,317,098; 5,371,241, 5,391,723; 5,416,203, 5,451,463; 5,510,475; 5,512,667; 5,514,785; 5,565,552; 5,567,810; 5,574,142; 5,585,481; 5,587,371; 5,595,726; 5,597,696; 5,599,923; 5,599,928; 5,688,941; 6,294,664; 6,320,017; 6,576,752; 6,783,931; 6,900,297; 7,037,646; and 8,106,022, the entire contents of each of which are hereby incorporated herein by reference. 
     It is not necessary for all positions in a given compound to be uniformly modified, and in fact more than one of the aforementioned modifications can be incorporated in a single compound or even at a single nucleoside within an iRNA. The present invention also includes iRNA compounds that are chimeric compounds. 
     “Chimeric” iRNA compounds or “chimeras,” in the context of this invention, are iRNA compounds, preferably dsRNA agents, that contain two or more chemically distinct regions, each made up of at least one monomer unit, i.e., a nucleotide in the case of a dsRNA compound. These iRNAs typically contain at least one region wherein the RNA is modified so as to confer upon the iRNA increased resistance to nuclease degradation, increased cellular uptake, or increased binding affinity for the target nucleic acid. An additional region of the iRNA can serve as a substrate for enzymes capable of cleaving RNA:DNA or RNA:RNA hybrids. By way of example, RNase H is a cellular endonuclease which cleaves the RNA strand of an RNA:DNA duplex. Activation of RNase H, therefore, results in cleavage of the RNA target, thereby greatly enhancing the efficiency of iRNA inhibition of gene expression. Consequently, comparable results can often be obtained with shorter iRNAs when chimeric dsRNAs are used, compared to phosphorothioate deoxy dsRNAs hybridizing to the same target region. Cleavage of the RNA target can be routinely detected by gel electrophoresis and, if necessary, associated nucleic acid hybridization techniques known in the art. 
     In certain instances, the RNA of an iRNA can be modified by a non-ligand group. A number of non-ligand molecules have been conjugated to iRNAs in order to enhance the activity, cellular distribution or cellular uptake of the iRNA, and procedures for performing such conjugations are available in the scientific literature. Such non-ligand moieties have included lipid moieties, such as cholesterol (Kubo, T. et al.,  Biochem. Biophys. Res. Comm.,  2007, 365(1):54-61; Letsinger et al.,  Proc. Natl. Acad. Sci. USA,  1989, 86:6553), cholic acid (Manoharan et al.,  Bioorg. Med. Chem. Lett.,  1994, 4:1053), a thioether, e.g., hexyl-S-tritylthiol (Manoharan et al.,  Ann. N.Y. Acad. Sci.,  1992, 660:306; Manoharan et al.,  Bioorg. Med. Chem. Let.,  1993, 3:2765), a thiocholesterol (Oberhauser et al.,  Nucl. Acids Res.,  1992, 20:533), an aliphatic chain, e.g., dodecandiol or undecyl residues (Saison-Behmoaras et al.,  EMBO J.,  1991, 10:111; Kabanov et al.,  FEBS Lett.,  1990, 259:327; Svinarchuk et al.,  Biochimie,  1993, 75:49), a phospholipid, e.g., di-hexadecyl-rac-glycerol or triethylammonium 1,2-di-O-hexadecyl-rac-glycero-3-H-phosphonate (Manoharan et al.,  Tetrahedron Lett.,  1995, 36:3651; Shea et al.,  Nucl. Acids Res.,  1990, 18:3777), a polyamine or a polyethylene glycol chain (Manoharan et al.,  Nucleosides  &amp;  Nucleotides,  1995, 14:969), or adamantane acetic acid (Manoharan et al.,  Tetrahedron Lett.,  1995, 36:3651), a palmityl moiety (Mishra et al.,  Biochim. Biophys. Acta,  1995, 1264:229), or an octadecylamine or hexylamino-carbonyl-oxycholesterol moiety (Crooke et al.,  J. Pharmacol. Exp. Ther.,  1996, 277:923). Representative United States patents that teach the preparation of such RNA conjugates have been listed above. Typical conjugation protocols involve the synthesis of RNAs bearing an aminolinker at one or more positions of the sequence. The amino group is then reacted with the molecule being conjugated using appropriate coupling or activating reagents. The conjugation reaction can be performed either with the RNA still bound to the solid support or following cleavage of the RNA, in solution phase. Purification of the RNA conjugate by HPLC typically affords the pure conjugate. 
     V. DELIVERY OF AN RNAI AGENT OF THE DISCLOSURE 
     The delivery of a RNAi agent of the disclosure to a cell e.g., a cell within a subject, such as a human subject (e.g., a subject in need thereof, such as a subject having an ACE2-associated disorder, e.g., a coronavirus-associated disorder, e.g., a subject having or at risk of developing of at risk of having a coronavirus infection, e.g., a subject having or at risk of developing or at risk of having Severe Acute Respiratory Syndrome 2 (SARS-CoV-2; COVID-19), Severe Acute Respiratory Syndrome (SARS-CoV), or Middle East Respiratory Syndrome (MERS-CoV)), can be achieved in a number of different ways. For example, delivery may be performed by contacting a cell with an RNAi agent of the disclosure either in vitro or in vivo. In vivo delivery may also be performed directly by administering a composition comprising an RNAi agent, e.g., a dsRNA, to a subject. Alternatively, in vivo delivery may be performed indirectly by administering one or more vectors that encode and direct the expression of the RNAi agent. These alternatives are discussed further below. 
     In general, any method of delivering a nucleic acid molecule (in vitro or in vivo) can be adapted for use with a RNAi agent of the disclosure (see e.g., Akhtar S. and Julian R L., (1992)  Trends Cell. Biol.  2(5):139-144 and WO94/02595, which are incorporated herein by reference in their entireties). For in vivo delivery, factors to consider in order to deliver an RNAi agent include, for example, biological stability of the delivered agent, prevention of non-specific effects, and accumulation of the delivered agent in the target tissue. The non-specific effects of an RNAi agent can be minimized by local administration, for example, by direct injection or implantation into a tissue or topically administering the preparation. Local administration to a treatment site maximizes local concentration of the agent, limits the exposure of the agent to systemic tissues that can otherwise be harmed by the agent or that can degrade the agent, and permits a lower total dose of the RNAi agent to be administered. Several studies have shown successful knockdown of gene products when an RNAi agent is administered locally. For example, pulmonary delivery, e.g., inhalation, of a dsRNA, e.g., SOD1, has been shown to effectively knockdown gene and protein expression in lung tissue and that there is excellent uptake of the dsRNA by the bronchioles and alveoli of the lung. Intraocular delivery of a VEGF dsRNA by intravitreal injection in cynomolgus monkeys (Tolentino, M J et al., (2004)  Retina  24:132-138) and subretinal injections in mice (Reich, S J. et al. (2003)  Mol. Vis.  9:210-216) were also both shown to prevent neovascularization in an experimental model of age-related macular degeneration. In addition, direct intratumoral injection of a dsRNA in mice reduces tumor volume (Pille, J. et al. (2005)  Mol. Ther.  11:267-274) and can prolong survival of tumor-bearing mice (Kim, W J. et al., (2006)  Mol. Ther.  14:343-350; Li, S. et al., (2007)  Mol. Ther.  15:515-523). RNA interference has also shown success with local delivery to the CNS by direct injection (Dorn, G. et al., (2004)  Nucleic Acids  32:e49; Tan, P H. et al. (2005)  Gene Ther.  12:59-66; Makimura, H. et al (2002)  BMC Neurosci.  3:18; Shishkina, G T., et al. (2004)  Neuroscience  129:521-528; Thakker, E R., et al. (2004)  Proc. Natl. Acad. Sci. U.S.A.  101:17270-17275; Akaneya, Y., et al. (2005)  J. Neurophysiol.  93:594-602) and to the lungs by intranasal administration (Howard, K A. et al., (2006)  Mol. Ther.  14:476-484; Zhang, X. et al., (2004)  J. Biol. Chem.  279:10677-10684; Bitko, V. et al., (2005)  Nat. Med.  11:50-55). For administering a RNAi agent systemically for the treatment of a disease, the RNA can be modified or alternatively delivered using a drug delivery system; both methods act to prevent the rapid degradation of the dsRNA by endo- and exo-nucleases in vivo. Modification of the RNA or the pharmaceutical carrier can also permit targeting of the RNAi agent to the target tissue and avoid undesirable off-target effects (e.g., without wishing to be bound by theory, use of GNAs as described herein has been identified to destabilize the seed region of a dsRNA, resulting in enhanced preference of such dsRNAs for on-target effectiveness, relative to off-target effects, as such off-target effects are significantly weakened by such seed region destabilization). RNAi agents can be modified by chemical conjugation to lipophilic groups such as cholesterol to enhance cellular uptake and prevent degradation. For example, a RNAi agent directed against ApoB conjugated to a lipophilic cholesterol moiety was injected systemically into mice and resulted in knockdown of apoB mRNA in both the liver and jejunum (Soutschek, J. et al., (2004)  Nature  432:173-178). Conjugation of an RNAi agent to an aptamer has been shown to inhibit tumor growth and mediate tumor regression in a mouse model of prostate cancer (McNamara, J O. et al., (2006)  Nat. Biotechnol.  24:1005-1015). In an alternative embodiment, the RNAi agent can be delivered using drug delivery systems such as a nanoparticle, a dendrimer, a polymer, liposomes, or a cationic delivery system. Positively charged cationic delivery systems facilitate binding of molecule RNAi agent (negatively charged) and also enhance interactions at the negatively charged cell membrane to permit efficient uptake of an RNAi agent by the cell. Cationic lipids, dendrimers, or polymers can either be bound to an RNAi agent, or induced to form a vesicle or micelle (see e.g., Kim S H. et al., (2008)  Journal of Controlled Release  129(2):107-116) that encases an RNAi agent. The formation of vesicles or micelles further prevents degradation of the RNAi agent when administered systemically. Methods for making and administering cationic-RNAi agent complexes are well within the abilities of one skilled in the art (see e.g., Sorensen, D R., et al. (2003)  J. Mol. Biol  327:761-766; Verma, U N. et al., (2003)  Clin. Cancer Res.  9:1291-1300; Arnold, A S et al. (2007)  J. Hypertens.  25:197-205, which are incorporated herein by reference in their entirety). Some non-limiting examples of drug delivery systems useful for systemic delivery of RNAi agents include DOTAP (Sorensen, D R., et al (2003), supra; Verma, U N. et al., (2003), supra), Oligofectamine, “solid nucleic acid lipid particles” (Zimmermann, T S. et al., (2006)  Nature  441:111-114), cardiolipin (Chien, P Y. et al., (2005)  Cancer Gene Ther.  12:321-328; Pal, A. et al., (2005)  Int J. Oncol.  26:1087-1091), polyethyleneimine (Bonnet M E. et al., (2008)  Pharm. Res . Aug. 16 Epub ahead of print; Aigner, A. (2006)  J. Biomed. Biotechnol.  71659), Arg-Gly-Asp (RGD) peptides (Liu, S. (2006)  Mol. Pharm.  3:472-487), and polyamidoamines (Tomalia, D A. et al., (2007)  Biochem. Soc. Trans.  35:61-67; Yoo, H. et al., (1999)  Pharm. Res.  16:1799-1804). In some embodiments, a RNAi agent forms a complex with cyclodextrin for systemic administration. Methods for administration and pharmaceutical compositions of RNAi agents and cyclodextrins can be found in U.S. Pat. No. 7,427,605, which is herein incorporated by reference in its entirety. 
     Certain aspects of the instant disclosure relate to a method of reducing the expression of an ACE2 gene in a cell, comprising contacting said cell with the double-stranded RNAi agent of the disclosure. In one embodiment, the cell is a hepatic cell, optionally a hepatocyte. In one embodiment, the cell is an extrahepatic cell, optionally a pulmonary cell. 
     Another aspect of the disclosure relates to a method of reducing the expression and/or activity of an ACE2 gene in a subject, comprising administering to the subject the double-stranded RNAi agent of the disclosure. 
     Another aspect of the disclosure relates to a method of treating a subject having an ACE2-associated disorder or at risk of having or at risk of developing an ACE2-associated disorder, comprising administering to the subject a therapeutically effective amount of the double-stranded RNAi agent of the disclosure, thereby treating the subject. In some embodiments, the ACE2-associated disorder comprises a coronavirus-associated disorder. Non-limiting examples of coronavirus-associated diseases include, for example, Severe Acute Respiratory Syndrome 2 (SARS-CoV-2; COVID-19), Severe Acute Respiratory Syndrome (SARS-CoV), or Middle East Respiratory Syndrome (MERS-CoV). 
     In one embodiment, the double-stranded RNAi agent is administered subcutaneously. 
     In one embodiment, the double-stranded RNAi agent is administered by pulmonary system administration, e.g., intranasal administration, or oral inhalative administration. 
     In one embodiment, the double-stranded RNAi agent is administered intranasally. 
     By pulmonary system administration, e.g., intranasal administration or oral inhalative administration, of the double-stranded RNAi agent, the method can reduce the expression of an ACE2 target gene in a pulmonary system tissue, e.g., a nasopharynx tissue, an oropharynx tissue, a laryngopharynx tissue, a larynx tissue, a trachea tissue, a carina tissue, a bronchi tissue, a bronchiole tissue, or an alveoli tissue. 
     For ease of exposition the formulations, compositions and methods in this section are discussed largely with regard to modified siRNA compounds. It may be understood, however, that these formulations, compositions and methods can be practiced with other siRNA compounds, e.g., unmodified siRNA compounds, and such practice is within the disclosure. A composition that includes a RNAi agent can be delivered to a subject by a variety of routes. Exemplary routes include pulmonary system, intravenous, intraventricular, topical, rectal, anal, vaginal, and ocular. 
     The RNAi agents of the disclosure can be incorporated into pharmaceutical compositions suitable for administration. Such compositions typically include one or more species of RNAi agent and a pharmaceutically acceptable carrier. As used herein the language “pharmaceutically acceptable carrier” is intended to include any and all solvents, dispersion media, coatings, antibacterial and antifungal agents, isotonic and absorption delaying agents, and the like, compatible with pharmaceutical administration. The use of such media and agents for pharmaceutically active substances is well known in the art. Except insofar as any conventional media or agent is incompatible with the active compound, use thereof in the compositions is contemplated. Supplementary active compounds can also be incorporated into the compositions. 
     The pharmaceutical compositions of the present disclosure may be administered in a number of ways depending upon whether local or systemic treatment is desired and upon the area to be treated. Administration may be pulmonary, e.g., by inhalation or insufflation of powders or aerosols, including by nebulizer; intratracheal, intranasal, topical (including ophthalmic, vaginal, rectal, intranasal, transdermal), oral, or parenteral. Parenteral administration includes intravenous drip, subcutaneous, intraperitoneal, or intramuscular injection, or intrathecal or intraventricular administration. 
     The route and site of administration may be chosen to enhance targeting. For example, to target muscle cells, intramuscular injection into the muscles of interest would be a logical choice. Lung cells might be targeted by administering the RNAi agent in powder or aerosol form. The vascular endothelial cells could be targeted by coating a balloon catheter with the RNAi agent and mechanically introducing the RNA. 
     Compositions for pulmonary system delivery may include aqueous solutions, e.g., for intranasal or oral inhalative administration, suitable carriers composed of, e.g., lipids (liposomes, niosomes, microemulsions, lipidic micelles, solid lipid nanoparticles) or polymers (polymer micelles, dendrimers, polymeric nanoparticles, nonogels, nanocapsules), adjuvant, e.g., for oral inhalative administration. Aqueous compositions may be sterile and may optionally contain buffers, diluents, absorbtion enhancers and other suitable additives. 
     Formulations for topical administration may include transdermal patches, ointments, lotions, creams, gels, drops, suppositories, sprays, liquids, and powders. Conventional pharmaceutical carriers, aqueous, powder or oily bases, thickeners and the like may be necessary or desirable. Coated condoms, gloves, and the like may also be useful. 
     Compositions for oral administration include powders or granules, suspensions or solutions in water, syrups, elixirs or non-aqueous media, tablets, capsules, lozenges, or troches. In the case of tablets, carriers that can be used include lactose, sodium citrate and salts of phosphoric acid. Various disintegrants such as starch, and lubricating agents such as magnesium stearate, sodium lauryl sulfate and talc, are commonly used in tablets. For oral administration in capsule form, useful diluents are lactose and high molecular weight polyethylene glycols. When aqueous suspensions are required for oral use, the nucleic acid compositions can be combined with emulsifying and suspending agents. If desired, certain sweetening or flavoring agents can be added. 
     Compositions for intrathecal or intraventricular administration may include sterile aqueous solutions which may also contain buffers, diluents, and other suitable additives. 
     Formulations for parenteral administration may include sterile aqueous solutions which may also contain buffers, diluents, and other suitable additives. Intraventricular injection may be facilitated by an intraventricular catheter, for example, attached to a reservoir. For intravenous use, the total concentration of solutes may be controlled to render the preparation isotonic. 
     In one embodiment, the administration of the siRNA compound, e.g., a double-stranded siRNA compound, or ssiRNA compound, composition is parenteral, e.g., intravenous (e.g., as a bolus or as a diffusible infusion), intradermal, intraperitoneal, intramuscular, intrathecal, intraventricular, intracranial, subcutaneous, transmucosal, buccal, sublingual, endoscopic, rectal, oral, vaginal, topical, pulmonary system, intranasal, urethral, or ocular. Administration can be provided by the subject or by another person, e.g., a health care provider. The medication can be provided in measured doses or in a dispenser which delivers a metered dose. Selected modes of delivery are discussed in more detail below. 
     Pulmonary System Administration 
     In one embodiment, the double-stranded RNAi agent is administered by pulmonary system administration. The pulmonary system includes the upper pulmonary system and the lower pulmonary system. The upper pulmonary system includes the nose and the pharynx. The pharynx includes the nasopharynx, oropharynx, and laryngopharynx. The lower pulmonary system includes the larynx, trachea, carina, bronchi, bronchioles, and alveoli. 
     Pulmonary system administration may be intranasal administration or oral inhalative administration. Such administration permits both systemic and local delivery of the double stranded RNAi agents of the invention. 
     Intranasal administration may include instilling or insufflating a double stranded RNAi agent into the nasal cavity with syringes or droppers by applying a few drops at a time or via atomization. 
     Suitable dosage forms for intranasal administration include drops, powders, nebulized mists, and sprays. 
     Oral inhalative administration may include use of device, e.g., a passive breath driven or active power driven single/-multiple dose dry powder inhaler (DPI), to deliver a double stranded RNAi agent to the pulmonary system. Suitable dosage forms for oral inhalative administration include powders and solutions. Suitable devices for oral inhalative administration include nebulizers, metered-dose inhalers, and dry powder inhalers. Dry powder inhalers are of the most popular devices used to deliver drugs, especially proteins to the lungs. Exemplary commercially available dry powder inhalers include Spinhaler (Fisons Pharmaceuticals, Rochester, N.Y.) and Rotahaler (GSK, RTP, NC). Several types of nebulizers are available, namely jet nebulizers, ultrasonic nebulizers, vibrating mesh nebulizers. Jet nebulizers are driven by compressed air. Ultrasonic nebulizers use a piezoelectric transducer in order to create droplets from an open liquid reservoir. Vibrating mesh nebulizers use perforated membranes actuated by an annular piezo element to vibrate in resonant bending mode. The holes in the membrane have a large cross-section size on the liquid supply side and a narrow cross-section size on the side from where the droplets emerge. Depending on the therapeutic application, the hole sizes and number of holes can be adjusted. Selection of a suitable device depends on parameters, such as nature of the drug and its formulation, the site of action, and pathophysiology of the lung. Aqueous suspensions and solutions are nebulized effectively. Aerosols based on mechanically generated vibration mesh technologies also have been used successfully to deliver proteins to lungs. 
     The amount of RNAi agent for pulmonary system administration may vary from one target gene to another target gene and the appropriate amount that has to be applied may have to be determined individually for each target gene. Typically, this amount ranges from 10 μg to 2 mg, preferably 50 μg to 1500 μg, more preferably 100 μg to 1000 μg. 
     Vector Encoded RNAi Agents of the Disclosure 
     RNAi agents targeting the ACE2 gene can be expressed from transcription units inserted into DNA or RNA vectors (see, e.g., Couture, A, et al.,  TIG . (1996), 12:5-10; WO 00/22113, WO 00/22114, and U.S. Pat. No. 6,054,299). Expression is preferably sustained (months or longer), depending upon the specific construct used and the target tissue or cell type. These transgenes can be introduced as a linear construct, a circular plasmid, or a viral vector, which can be an integrating or non-integrating vector. The transgene can also be constructed to permit it to be inherited as an extrachromosomal plasmid (Gassmann, et al., (1995)  Proc. Natl. Acad. Sci. USA  92:1292). 
     The individual strand or strands of a RNAi agent can be transcribed from a promoter on an expression vector. Where two separate strands are to be expressed to generate, for example, a dsRNA, two separate expression vectors can be co-introduced (e.g., by transfection or infection) into a target cell. Alternatively, each individual strand of a dsRNA can be transcribed by promoters both of which are located on the same expression plasmid. In one embodiment, a dsRNA is expressed as inverted repeat polynucleotides joined by a linker polynucleotide sequence such that the dsRNA has a stem and loop structure. 
     RNAi agent expression vectors are generally DNA plasmids or viral vectors. Expression vectors compatible with eukaryotic cells, preferably those compatible with vertebrate cells, can be used to produce recombinant constructs for the expression of a RNAi agent as described herein. Delivery of RNAi agent expressing vectors can be systemic, such as by intravenous or intramuscular administration, by administration to target cells ex-planted from the patient followed by reintroduction into the patient, or by any other means that allows for introduction into a desired target cell. 
     Viral vector systems which can be utilized with the methods and compositions described herein include, but are not limited to, (a) adenovirus vectors; (b) retrovirus vectors, including but not limited to lentiviral vectors, moloney murine leukemia virus, etc.; (c) adeno-associated virus vectors; (d) herpes simplex virus vectors; (e) SV 40 vectors; (f) polyoma virus vectors; (g) papilloma virus vectors; (h) picornavirus vectors; (i) pox virus vectors such as an orthopox, e.g., vaccinia virus vectors or avipox, e.g. canary pox or fowl pox; and (j) a helper-dependent or gutless adenovirus. Replication-defective viruses can also be advantageous. Different vectors will or will not become incorporated into the cells&#39; genome. The constructs can include viral sequences for transfection, if desired. Alternatively, the construct can be incorporated into vectors capable of episomal replication, e.g. EPV and EBV vectors. Constructs for the recombinant expression of a RNAi agent will generally require regulatory elements, e.g., promoters, enhancers, etc., to ensure the expression of the RNAi agent in target cells. Other aspects to consider for vectors and constructs are known in the art. 
     VI. PHARMACEUTICAL COMPOSITIONS OF THE INVENTION 
     The present disclosure also includes pharmaceutical compositions and formulations which include the RNAi agents of the disclosure. In one embodiment, provided herein are pharmaceutical compositions containing an RNAi agent, as described herein, and a pharmaceutically acceptable carrier. The pharmaceutical compositions containing the RNAi agent are useful for treating a subject who would benefit from inhibiting or reducing the expression of an ACE2 gene, e.g., a subject having an ACE2-associated disorder, e.g, a coronavirus-associated disorder, e.g., a subject having or at risk of having or at risk of developing a coronavirus infection, e.g., a subject having Severe Acute Respiratory Syndrome 2 (SARS-CoV-2; COVID-19), Severe Acute Respiratory Syndrome (SARS-CoV), or Middle East Respiratory Syndrome (MERS-CoV). Such pharmaceutical compositions are formulated based on the mode of delivery. One example is compositions that are formulated for direct delivery into the the pulmonary system by intranasal administration or oral inhalative administration, e.g., by inhalation or insufflation of powders or aerosols, including by nebulizer; intratracheal or intranasal delivery. Another example is compositions that are formulated for systemic administration via parenteral delivery, e.g., by intravenous (IV), intramuscular (IM), or for subcutaneous (subQ) delivery. 
     In some embodiments, the pharmaceutical compositions of the invention are pyrogen free or non-pyrogenic. 
     The pharmaceutical compositions of the disclosure may be administered in dosages sufficient to inhibit expression of an ACE2 gene. In general, a suitable dose of an RNAi agent of the disclosure will be a flat dose in the range of about 0.001 to about 200.0 mg about once per month to about once per year, typically about once per quarter (i.e., about once every three months) to about once per year, generally a flat dose in the range of about 1 to 50 mg about once per month to about once per year, typically about once per quarter to about once per year. 
     After an initial treatment regimen (e.g., loading dose), the treatments can be administered on a less frequent basis. 
     The skilled artisan will appreciate that certain factors can influence the dosage and timing required to effectively treat a subject, including but not limited to the severity of the disease or disorder, previous treatments, the general health or age of the subject, and other diseases present. Moreover, treatment of a subject with a therapeutically effective amount of a composition can include a single treatment or a series of treatments. 
     Advances in mouse genetics have generated a number of mouse models for the study of various ACE2-associated diseases that would benefit from reduction in the expression of ACE2. Such models can be used for in vivo testing of RNAi agents, as well as for determining a therapeutically effective dose. Suitable mouse models are known in the art and include, for example, the mouse models described elsewhere herein. 
     The pharmaceutical compositions of the present disclosure can be administered in a number of ways depending upon whether local or systemic treatment is desired and upon the area to be treated. Administration can be topical (e.g., by a transdermal patch), pulmonary system administration by intranasal administration or oral inhalative administration, e.g., by inhalation or insufflation of powders or aerosols, including by nebulizer; intratracheal, epidermal and transdermal, oral or parenteral. Parenteral administration includes intravenous, intraarterial, subcutaneous, intraperitoneal or intramuscular injection or infusion; subdermal, e.g., via an implanted device; or intracranial, e.g., by intraparenchymal, intrathecal or intraventricular, administration. 
     The RNAi agents can be delivered in a manner to target a particular tissue, such as the liver, the lung (e.g., bronchioles, alveoli, or bronchus of the lung), or both the liver and lung. 
     Pharmaceutical compositions and formulations for topical administration can include transdermal patches, ointments, lotions, creams, gels, drops, suppositories, sprays, liquids and powders. Conventional pharmaceutical carriers, aqueous, powder or oily bases, thickeners and the like can be necessary or desirable. Coated condoms, gloves and the like can also be useful. Suitable topical formulations include those in which the RNAi agents featured in the disclosure are in admixture with a topical delivery agent such as lipids, liposomes, fatty acids, fatty acid esters, steroids, chelating agents and surfactants. Suitable lipids and liposomes include neutral (e.g., dioleoylphosphatidyl DOPE ethanolamine, dimyristoylphosphatidyl choline DMPC, distearolyphosphatidyl choline) negative (e.g., dimyristoylphosphatidyl glycerol DMPG) and cationic (e.g., dioleoyltetramethylaminopropyl DOTAP and dioleoylphosphatidyl ethanolamine DOTMA). RNAi agents featured in the disclosure can be encapsulated within liposomes or can form complexes thereto, in particular to cationic liposomes. Alternatively, RNAi agents can be complexed to lipids, in particular to cationic lipids. Suitable fatty acids and esters include but are not limited to arachidonic acid, oleic acid, eicosanoic acid, lauric acid, caprylic acid, capric acid, myristic acid, palmitic acid, stearic acid, linoleic acid, linolenic acid, dicaprate, tricaprate, monoolein, dilaurin, glyceryl 1-monocaprate, 1-dodecylazacycloheptan-2-one, an acylcarnitine, an acylcholine, or a C 1-20  alkyl ester (e.g., isopropylmyristate IPM), monoglyceride, diglyceride or pharmaceutically acceptable salt thereof. Topical formulations are described in detail in U.S. Pat. No. 6,747,014, which is incorporated herein by reference. 
     A. RNAi Agent Formulations Comprising Membranous Molecular Assemblies 
     A RNAi agent for use in the compositions and methods of the disclosure can be formulated for delivery in a membranous molecular assembly, e.g., a liposome or a micelle. As used herein, the term “liposome” refers to a vesicle composed of amphiphilic lipids arranged in at least one bilayer, e.g., one bilayer or a plurality of bilayers. Liposomes include unilamellar and multilamellar vesicles that have a membrane formed from a lipophilic material and an aqueous interior. The aqueous portion contains the RNAi agent composition. The lipophilic material isolates the aqueous interior from an aqueous exterior, which typically does not include the RNAi agent composition, although in some examples, it may. Liposomes are useful for the transfer and delivery of active ingredients to the site of action. Because the liposomal membrane is structurally similar to biological membranes, when liposomes are applied to a tissue, the liposomal bilayer fuses with bilayer of the cellular membranes. As the merging of the liposome and cell progresses, the internal aqueous contents that include the RNAi agent are delivered into the cell where the RNAi agent can specifically bind to a target RNA and can mediate RNAi. In some cases the liposomes are also specifically targeted, e.g., to direct the RNAi agent to particular cell types. 
     A liposome containing an RNAi agent can be prepared by a variety of methods. In one example, the lipid component of a liposome is dissolved in a detergent so that micelles are formed with the lipid component. For example, the lipid component can be an amphipathic cationic lipid or lipid conjugate. The detergent can have a high critical micelle concentration and may be nonionic. Exemplary detergents include cholate, CHAPS, octylglucoside, deoxycholate, and lauroyl sarcosine. The RNAi agent preparation is then added to the micelles that include the lipid component. The cationic groups on the lipid interact with the RNAi agent and condense around the RNAi agent to form a liposome. After condensation, the detergent is removed, e.g., by dialysis, to yield a liposomal preparation of RNAi agent. 
     If necessary a carrier compound that assists in condensation can be added during the condensation reaction, e.g., by controlled addition. For example, the carrier compound can be a polymer other than a nucleic acid (e.g., spermine or spermidine). pH can also adjusted to favor condensation. 
     Methods for producing stable polynucleotide delivery vehicles, which incorporate a polynucleotide/cationic lipid complex as structural components of the delivery vehicle, are further described in, e.g., WO 96/37194, the entire contents of which are incorporated herein by reference. Liposome formation can also include one or more aspects of exemplary methods described in Felgner, P. L. et al., (1987)  Proc. Natl. Acad. Sci. USA  8:7413-7417; U.S. Pat. Nos. 4,897,355; 5,171,678; Bangham et al., (1965)  M. Mol. Biol.  23:238; Olson et al., (1979)  Biochim. Biophys. Acta  557:9; Szoka et al., (1978)  Proc. Natl. Acad. Sci.  75: 4194; Mayhew et al., (1984)  Biochim. Biophys. Acta  775:169; Kim et al., (1983)  Biochim. Biophys. Acta  728:339; and Fukunaga et al., (1984)  Endocrinol.  115:757. Commonly used techniques for preparing lipid aggregates of appropriate size for use as delivery vehicles include sonication and freeze-thaw plus extrusion (see, e.g., Mayer et al., (1986)  Biochim. Biophys. Acta  858:161. Microfluidization can be used when consistently small (50 to 200 nm) and relatively uniform aggregates are desired (Mayhew et al., (1984)  Biochim. Biophys. Acta  775:169. These methods are readily adapted to packaging RNAi agent preparations into liposomes. 
     Liposomes fall into two broad classes. Cationic liposomes are positively charged liposomes which interact with the negatively charged nucleic acid molecules to form a stable complex. The positively charged nucleic acid/liposome complex binds to the negatively charged cell surface and is internalized in an endosome. Due to the acidic pH within the endosome, the liposomes are ruptured, releasing their contents into the cell cytoplasm (Wang et al. (1987)  Biochem. Biophys. Res. Commun.,  147:980-985). 
     Liposomes, which are pH-sensitive or negatively charged, entrap nucleic acids rather than complex with them. Since both the nucleic acid and the lipid are similarly charged, repulsion rather than complex formation occurs. Nevertheless, some nucleic acid is entrapped within the aqueous interior of these liposomes. pH sensitive liposomes have been used to deliver nucleic acids encoding the thymidine kinase gene to cell monolayers in culture. Expression of the exogenous gene was detected in the target cells (Zhou et al. (1992)  Journal of Controlled Release,  19:269-274). 
     One major type of liposomal composition includes phospholipids other than naturally-derived phosphatidylcholine. Neutral liposome compositions, for example, can be formed from dimyristoyl phosphatidylcholine (DMPC) or dipalmitoyl phosphatidylcholine (DPPC). Anionic liposome compositions generally are formed from dimyristoyl phosphatidylglycerol, while anionic fusogenic liposomes are formed primarily from dioleoyl phosphatidylethanolamine (DOPE). Another type of liposomal composition is formed from phosphatidylcholine (PC) such as, for example, soybean PC, and egg PC. Another type is formed from mixtures of phospholipid or phosphatidylcholine or cholesterol. 
     Examples of other methods to introduce liposomes into cells in vitro and in vivo include U.S. Pat. Nos. 5,283,185; 5,171,678; WO 94/00569; WO 93/24640; WO 91/16024; Felgner, (1994)  J. Biol. Chem.  269:2550; Nabel, (1993)  Proc. Natl. Acad. Sci.  90:11307; Nabel, (1992)  Human Gene Ther.  3:649; Gershon, (1993)  Biochem.  32:7143; and Strauss, (1992)  EMBO J.  11:417. 
     Non-ionic liposomal systems have also been examined to determine their utility in the delivery of drugs to the skin, in particular systems comprising non-ionic surfactant and cholesterol. Non-ionic liposomal formulations comprising Novasome™ I (glyceryl dilaurate/cholesterol/polyoxyethylene-10-stearyl ether) and Novasome™ II (glyceryl distearate/cholesterol/polyoxyethylene-10-stearyl ether) were used to deliver cyclosporin-A into the dermis of mouse skin. Results indicated that such non-ionic liposomal systems were effective in facilitating the deposition of cyclosporine A into different layers of the skin (Hu et al., (1994)  S.T.P. Pharma. Sci.,  4(6):466). 
     Liposomes also include “sterically stabilized” liposomes, a term which, as used herein, refers to liposomes comprising one or more specialized lipids that, when incorporated into liposomes, result in enhanced circulation lifetimes relative to liposomes lacking such specialized lipids. Examples of sterically stabilized liposomes are those in which part of the vesicle-forming lipid portion of the liposome (A) comprises one or more glycolipids, such as monosialoganglioside G M1 , or (B) is derivatized with one or more hydrophilic polymers, such as a polyethylene glycol (PEG) moiety. While not wishing to be bound by any particular theory, it is thought in the art that, at least for sterically stabilized liposomes containing gangliosides, sphingomyelin, or PEG-derivatized lipids, the enhanced circulation half-life of these sterically stabilized liposomes derives from a reduced uptake into cells of the reticuloendothelial system (RES) (Allen et al., (1987)  FEBS Letters,  223:42; Wu et al., (1993)  Cancer Research,  53:3765). 
     Various liposomes comprising one or more glycolipids are known in the art. Papahadjopoulos et al. ( Ann. N.Y. Acad. Sci.,  (1987), 507:64) reported the ability of monosialoganglioside G M1 , galactocerebroside sulfate and phosphatidylinositol to improve blood half-lives of liposomes. These findings were expounded upon by Gabizon et al. ( Proc. Natl. Acad. Sci. U.S.A.,  (1988), 85, 6949). U.S. Pat. No. 4,837,028 and WO 88/04924, both to Allen et al., disclose liposomes comprising (1) sphingomyelin and (2) the ganglioside G M1  or a galactocerebroside sulfate ester. U.S. Pat. No. 5,543,152 (Webb et al.) discloses liposomes comprising sphingomyelin. Liposomes comprising 1,2-sn-dimyristoylphosphatidylcholine are disclosed in WO 97/13499 (Lim et al). 
     In one embodiment, cationic liposomes are used. Cationic liposomes possess the advantage of being able to fuse to the cell membrane. Non-cationic liposomes, although not able to fuse as efficiently with the plasma membrane, are taken up by macrophages in vivo and can be used to deliver RNAi agents to macrophages. 
     Further advantages of liposomes include: liposomes obtained from natural phospholipids are biocompatible and biodegradable; liposomes can incorporate a wide range of water and lipid soluble drugs; liposomes can protect encapsulated RNAi agents in their internal compartments from metabolism and degradation (Rosoff, in “Pharmaceutical Dosage Forms,” Lieberman, Rieger and Banker (Eds.), 1988, volume 1, p. 245). Important considerations in the preparation of liposome formulations are the lipid surface charge, vesicle size and the aqueous volume of the liposomes. 
     A positively charged synthetic cationic lipid, N-[1-(2,3-dioleyloxy)propyl]-N,N,N-trimethylammonium chloride (DOTMA) can be used to form small liposomes that interact spontaneously with nucleic acid to form lipid-nucleic acid complexes which are capable of fusing with the negatively charged lipids of the cell membranes of tissue culture cells, resulting in delivery of RNAi agent (see, e.g., Felgner, P. L. et al., (1987)  Proc. Natl. Acad. Sci. USA  8:7413-7417, and U.S. Pat. No. 4,897,355 for a description of DOTMA and its use with DNA). 
     A DOTMA analogue, 1,2-bis(oleoyloxy)-3-(trimethylammonia)propane (DOTAP) can be used in combination with a phospholipid to form DNA-complexing vesicles. Lipofectin™ Bethesda Research Laboratories, Gaithersburg, Md.) is an effective agent for the delivery of highly anionic nucleic acids into living tissue culture cells that comprise positively charged DOTMA liposomes which interact spontaneously with negatively charged polynucleotides to form complexes. When enough positively charged liposomes are used, the net charge on the resulting complexes is also positive. Positively charged complexes prepared in this way spontaneously attach to negatively charged cell surfaces, fuse with the plasma membrane, and efficiently deliver functional nucleic acids into, for example, tissue culture cells. Another commercially available cationic lipid, 1,2-bis(oleoyloxy)-3,3-(trimethylammonia)propane (“DOTAP”) (Boehringer Mannheim, Indianapolis, Ind.) differs from DOTMA in that the oleoyl moieties are linked by ester, rather than ether linkages. 
     Other reported cationic lipid compounds include those that have been conjugated to a variety of moieties including, for example, carboxyspermine which has been conjugated to one of two types of lipids and includes compounds such as 5-carboxyspermylglycine dioctaoleoylamide (“DOGS”) (Transfectam™, Promega, Madison, Wis.) and dipalmitoylphosphatidylethanolamine 5-carboxyspermyl-amide (“DPPES”) (see, e.g., U.S. Pat. No. 5,171,678). 
     Another cationic lipid conjugate includes derivatization of the lipid with cholesterol (“DC-Chol”) which has been formulated into liposomes in combination with DOPE (See, Gao, X. and Huang, L., (1991)  Biochim. Biophys. Res. Commun.  179:280). Lipopolylysine, made by conjugating polylysine to DOPE, has been reported to be effective for transfection in the presence of serum (Zhou, X. et al., (1991)  Biochim. Biophys. Acta  1065:8). For certain cell lines, these liposomes containing conjugated cationic lipids, are said to exhibit lower toxicity and provide more efficient transfection than the DOTMA-containing compositions. Other commercially available cationic lipid products include DMRIE and DMRIE-HP (Vical, La Jolla, Calif.) and Lipofectamine (DOSPA) (Life Technology, Inc., Gaithersburg, Md.). Other cationic lipids suitable for the delivery of oligonucleotides are described in WO 98/39359 and WO 96/37194. 
     Liposomal formulations are particularly suited for topical administration, liposomes present several advantages over other formulations. Such advantages include reduced side effects related to high systemic absorption of the administered drug, increased accumulation of the administered drug at the desired target, and the ability to administer RNAi agent into the skin. In some implementations, liposomes are used for delivering RNAi agent to epidermal cells and also to enhance the penetration of RNAi agent into dermal tissues, e.g., into skin. For example, the liposomes can be applied topically. Topical delivery of drugs formulated as liposomes to the skin has been documented (see, e.g., Weiner et al., (1992)  Journal of Drug Targeting , vol. 2, 405-410 and du Plessis et al., (1992)  Antiviral Research,  18:259-265; Mannino, R. J. and Fould-Fogerite, S., (1998)  Biotechniques  6:682-690; Itani, T. et al., (1987)  Gene  56:267-276; Nicolau, C. et al. (1987)  Meth. Enzymol.  149:157-176; Straubinger, R. M. and Papahadjopoulos, D. (1983)  Meth. Enzymol.  101:512-527; Wang, C. Y. and Huang, L., (1987)  Proc. Natl. Acad. Sci. USA  84:7851-7855). 
     Non-ionic liposomal systems have also been examined to determine their utility in the delivery of drugs to the skin, in particular systems comprising non-ionic surfactant and cholesterol. Non-ionic liposomal formulations comprising Novasome I (glyceryl dilaurate/cholesterol/polyoxyethylene-10-stearyl ether) and Novasome II (glyceryl distearate/cholesterol/polyoxyethylene-10-stearyl ether) were used to deliver a drug into the dermis of mouse skin. Such formulations with RNAi agent are useful for treating a dermatological disorder. 
     Liposomes that include RNAi agents can be made highly deformable. Such deformability can enable the liposomes to penetrate through pore that are smaller than the average radius of the liposome. For example, transfersomes are a type of deformable liposomes. Transfersomes can be made by adding surface edge activators, usually surfactants, to a standard liposomal composition. Transfersomes that include RNAi agent can be delivered, for example, subcutaneously by infection in order to deliver RNAi agent to keratinocytes in the skin. In order to cross intact mammalian skin, lipid vesicles must pass through a series of fine pores, each with a diameter less than 50 nm, under the influence of a suitable transdermal gradient. In addition, due to the lipid properties, these transfersomes can be self-optimizing (adaptive to the shape of pores, e.g., in the skin), self-repairing, and can frequently reach their targets without fragmenting, and often self-loading. 
     Other formulations amenable to the present disclosure are described in PCT publication No. WO 2008/042973. 
     Transfersomes, yet another type of liposomes, are highly deformable lipid aggregates which are attractive candidates for drug delivery vehicles. Transfersomes can be described as lipid droplets which are so highly deformable that they are easily able to penetrate through pores which are smaller than the droplet. Transfersomes are adaptable to the environment in which they are used, e.g., they are self-optimizing (adaptive to the shape of pores in the skin), self-repairing, frequently reach their targets without fragmenting, and often self-loading. To make transfersomes it is possible to add surface edge-activators, usually surfactants, to a standard liposomal composition. Transfersomes have been used to deliver serum albumin to the skin. The transfersomes-mediated delivery of serum albumin has been shown to be as effective as subcutaneous injection of a solution containing serum albumin. 
     Surfactants find wide application in formulations such as those described herein, particularly in emulsions (including microemulsions) and liposomes. The most common way of classifying and ranking the properties of the many different types of surfactants, both natural and synthetic, is by the use of the hydrophile/lipophile balance (HLB). The nature of the hydrophilic group (also known as the “head”) provides the most useful means for categorizing the different surfactants used in formulations (Rieger, in Pharmaceutical Dosage Forms, Marcel Dekker, Inc., New York, N.Y., 1988, p. 285). 
     If the surfactant molecule is not ionized, it is classified as a nonionic surfactant. Nonionic surfactants find wide application in pharmaceutical and cosmetic products and are usable over a wide range of pH values. In general, their HLB values range from 2 to about 18 depending on their structure. Nonionic surfactants include nonionic esters such as ethylene glycol esters, propylene glycol esters, glyceryl esters, polyglyceryl esters, sorbitan esters, sucrose esters, and ethoxylated esters. Nonionic alkanolamides and ethers such as fatty alcohol ethoxylates, propoxylated alcohols, and ethoxylated/propoxylated block polymers are also included in this class. The polyoxyethylene surfactants are the most popular members of the nonionic surfactant class. 
     If the surfactant molecule carries a negative charge when it is dissolved or dispersed in water, the surfactant is classified as anionic. Anionic surfactants include carboxylates such as soaps, acyl lactylates, acyl amides of amino acids, esters of sulfuric acid such as alkyl sulfates and ethoxylated alkyl sulfates, sulfonates such as alkyl benzene sulfonates, acyl isethionates, acyl taurates and sulfosuccinates, and phosphates. The most important members of the anionic surfactant class are the alkyl sulfates and the soaps. 
     If the surfactant molecule carries a positive charge when it is dissolved or dispersed in water, the surfactant is classified as cationic. Cationic surfactants include quaternary ammonium salts and ethoxylated amines. The quaternary ammonium salts are the most used members of this class. 
     If the surfactant molecule has the ability to carry either a positive or negative charge, the surfactant is classified as amphoteric. Amphoteric surfactants include acrylic acid derivatives, substituted alkylamides, N-alkylbetaines and phosphatides. 
     The use of surfactants in drug products, formulations and in emulsions has been reviewed (Rieger, in Pharmaceutical Dosage Forms, Marcel Dekker, Inc., New York, N.Y., 1988, p. 285). 
     The RNAi agent for use in the methods of the disclosure can also be provided as micellar formulations. “Micelles” are defined herein as a particular type of molecular assembly in which amphipathic molecules are arranged in a spherical structure such that all the hydrophobic portions of the molecules are directed inward, leaving the hydrophilic portions in contact with the surrounding aqueous phase. The converse arrangement exists if the environment is hydrophobic. 
     A mixed micellar formulation suitable for delivery through transdermal membranes may be prepared by mixing an aqueous solution of the siRNA composition, an alkali metal C 8  to C22 alkyl sulphate, and a micelle forming compounds. Exemplary micelle forming compounds include lecithin, hyaluronic acid, pharmaceutically acceptable salts of hyaluronic acid, glycolic acid, lactic acid, chamomile extract, cucumber extract, oleic acid, linoleic acid, linolenic acid, monoolein, monooleates, monolaurates, borage oil, evening of primrose oil, menthol, trihydroxy oxo cholanyl glycine and pharmaceutically acceptable salts thereof, glycerin, polyglycerin, lysine, polylysine, triolein, polyoxyethylene ethers and analogues thereof, polidocanol alkyl ethers and analogues thereof, chenodeoxycholate, deoxycholate, and mixtures thereof. The micelle forming compounds may be added at the same time or after addition of the alkali metal alkyl sulphate. Mixed micelles will form with substantially any kind of mixing of the ingredients but vigorous mixing in order to provide smaller size micelles. 
     In one method a first micellar composition is prepared which contains the siRNA composition and at least the alkali metal alkyl sulphate. The first micellar composition is then mixed with at least three micelle forming compounds to form a mixed micellar composition. In another method, the micellar composition is prepared by mixing the siRNA composition, the alkali metal alkyl sulphate and at least one of the micelle forming compounds, followed by addition of the remaining micelle forming compounds, with vigorous mixing. 
     Phenol or m-cresol may be added to the mixed micellar composition to stabilize the formulation and protect against bacterial growth. Alternatively, phenol or m-cresol may be added with the micelle forming ingredients. An isotonic agent such as glycerin may also be added after formation of the mixed micellar composition. 
     For delivery of the micellar formulation as a spray, the formulation can be put into an aerosol dispenser and the dispenser is charged with a propellant. The propellant, which is under pressure, is in liquid form in the dispenser. The ratios of the ingredients are adjusted so that the aqueous and propellant phases become one, i.e., there is one phase. If there are two phases, it is necessary to shake the dispenser prior to dispensing a portion of the contents, e.g., through a metered valve. The dispensed dose of pharmaceutical agent is propelled from the metered valve in a fine spray. 
     Propellants may include hydrogen-containing chlorofluorocarbons, hydrogen-containing fluorocarbons, dimethyl ether and diethyl ether. In certain embodiments, HFA 134a (1,1,1,2 tetrafluoroethane) may be used. 
     The specific concentrations of the essential ingredients can be determined by relatively straightforward experimentation. For absorption through the oral cavities, it is often desirable to increase, e.g., at least double or triple, the dosage for through injection or administration through the gastrointestinal tract. 
     Lipid Particles 
     RNAi agents, e.g., dsRNAs of in the disclosure may be fully encapsulated in a lipid formulation, e.g., a LNP, or other nucleic acid-lipid particle. 
     As used herein, the term “LNP” refers to a stable nucleic acid-lipid particle. LNPs typically contain a cationic lipid, a non-cationic lipid, and a lipid that prevents aggregation of the particle (e.g., a PEG-lipid conjugate). LNPs are extremely useful for systemic applications, as they exhibit extended circulation lifetimes following intravenous (i.v.) injection and accumulate at distal sites (e.g., sites physically separated from the administration site). LNPs include “pSPLP,” which include an encapsulated condensing agent-nucleic acid complex as set forth in WO 00/03683. The particles of the present disclosure typically have a mean diameter of about 50 nm to about 150 nm, more typically about 60 nm to about 130 nm, more typically about 70 nm to about 110 nm, most typically about 70 nm to about 90 nm, and are substantially nontoxic. In addition, the nucleic acids when present in the nucleic acid-lipid particles of the present disclosure are resistant in aqueous solution to degradation with a nuclease. Nucleic acid-lipid particles and their method of preparation are disclosed in, e.g., U.S. Pat. Nos. 5,976,567; 5,981,501; 6,534,484; 6,586,410; 6,815,432; United States Patent publication No. 2010/0324120 and WO 96/40964. 
     In one embodiment, the lipid to drug ratio (mass/mass ratio) (e.g., lipid to dsRNA ratio) will be in the range of from about 1:1 to about 50:1, from about 1:1 to about 25:1, from about 3:1 to about 15:1, from about 4:1 to about 10:1, from about 5:1 to about 9:1, or about 6:1 to about 9:1. Ranges intermediate to the above recited ranges are also contemplated to be part of the disclosure. 
     Certain specific LNP formulations for delivery of RNAi agents have been described in the art, including, e.g., “LNP01” formulations as described in, e.g., WO 2008/042973, which is hereby incorporated by reference. 
     Additional exemplary lipid-dsRNA formulations are identified in the table below. 
     
       
         
           
               
               
               
             
               
                   
                   
               
               
                   
                   
                 cationic lipid/non-cationic 
               
               
                   
                   
                 lipid/cholesterol/PEG-lipid 
               
               
                   
                   
                 conjugate 
               
               
                   
                 Ionizable/Cationic Lipid 
                 Lipid:siRNA ratio 
               
               
                   
                   
               
             
            
               
                   
               
            
           
           
               
               
               
            
               
                 SNALP-1 
                 1,2-Dilinolenyloxy-N,N- 
                 DLinDMA/DPPC/Cholesterol/PEG- 
               
               
                   
                 dimethylaminopropane (DLinDMA) 
                 cDMA (57.1/7.1/34.4/1.4) 
               
               
                   
                   
                 lipid:siRNA ~7:1 
               
               
                 2-XTC 
                 2,2-Dilinoleyl-4-dimethylaminoethyl-[1,3]- 
                 XTC/DPPC/Cholesterol/PEG- 
               
               
                   
                 dioxolane (XTC) 
                 cDMA 57.1/7.1/34.4/1.4 
               
               
                   
                   
                 lipid:siRNA ~7:1 
               
               
                 LNP05 
                 2,2-Dilinoleyl-4-dimethylaminoethyl-[1,3]- 
                 XTC/DSPC/Cholesterol/PEG-DMG 
               
               
                   
                 dioxolane (XTC) 
                 57.5/7.5/31.5/3.5 
               
               
                   
                   
                 lipid:siRNA ~6:1 
               
               
                 LNP06 
                 2,2-Dilinoleyl-4-dimethylaminoethyl-[1,3]- 
                 XTC/DSPC/Cholesterol/PEG-DMG 
               
               
                   
                 dioxolane (XTC) 
                 57.5/7.5/31.5/3.5 
               
               
                   
                   
                 lipid:siRNA ~11:1 
               
               
                 LNP07 
                 2,2-Dilinoleyl-4-dimethylaminoethyl-[1,3]- 
                 XTC/DSPC/Cholesterol/PEG-DMG 
               
               
                   
                 dioxolane (XTC) 
                 60/7.5/31/1.5, 
               
               
                   
                   
                 lipid:siRNA ~6:1 
               
               
                 LNP08 
                 2,2-Dilinoleyl-4-dimethylaminoethyl-[1,3]- 
                 XTC/DSPC/Cholesterol/PEG-DMG 
               
               
                   
                 dioxolane (XTC) 
                 60/7.5/31/1.5, 
               
               
                   
                   
                 lipid:siRNA ~11:1 
               
               
                 LNP09 
                 2,2-Dilinoleyl-4-dimethylaminoethyl-[1,3]- 
                 XTC/DSPC/Cholesterol/PEG-DMG 
               
               
                   
                 dioxolane (XTC) 
                 50/10/38.5/1.5 
               
               
                   
                   
                 Lipid:siRNA 10:1 
               
               
                 LNP10 
                 (3aR,5s,6aS)-N,N-dimethyl-2,2-di((9Z,12Z)- 
                 ALN100/DSPC/Cholesterol/PEG-DMG 
               
               
                   
                 octadeca-9,12-dienyl)tetrahydro-3aH- 
                 50/10/38.5/1.5 
               
               
                   
                 cyclopenta[d][1,3]dioxol-5-amine (ALN100) 
                 Lipid:siRNA 10:1 
               
               
                 LNP11 
                 (6Z,9Z,28Z,31Z)-heptatriaconta-6,9,28,31- 
                 MC-3/DSPC/Cholesterol/PEG- 
               
               
                   
                 tetraen-19-yl 4-(dimethylamino)butanoate 
                 DMG 50/10/38.5/1.5 
               
               
                   
                 (MC3) 
                 Lipid:siRNA 10:1 
               
               
                 LNP12 
                 1,1′-(2-(4-(2-((2-(bis(2- 
                 Tech G1/DSPC/Cholesterol/PEG- 
               
               
                   
                 hydroxydodecyl)amino)ethyl)(2- 
                 DMG 50/10/38.5/1.5 
               
               
                   
                 hydroxydodecyl)amino)ethyl)piperazin-1- 
                 Lipid:siRNA 10:1 
               
               
                   
                 yl)ethylazanediyl)didodecan-2-ol (Tech G1) 
               
               
                 LNP13 
                 XTC 
                 XTC/DSPC/Chol/PEG-DMG 
               
               
                   
                   
                 50/10/38.5/1.5 
               
               
                   
                   
                 Lipid:siRNA: 33:1 
               
               
                 LNP14 
                 MC3 
                 MC3/DSPC/Chol/PEG-DMG 
               
               
                   
                   
                 40/15/40/5 
               
               
                   
                   
                 Lipid:siRNA: 11:1 
               
               
                 LNP15 
                 MC3 
                 MC3/DSPC/Chol/PEG- 
               
               
                   
                   
                 DSG/GalNAc-PEG-DSG 
               
               
                   
                   
                 50/10/35/4.5/0.5 
               
               
                   
                   
                 Lipid:siRNA: 11:1 
               
               
                 LNP16 
                 MC3 
                 MC3/DSPC/Chol/PEG-DMG 
               
               
                   
                   
                 50/10/38.5/1.5 
               
               
                   
                   
                 Lipid:siRNA: 7:1 
               
               
                 LNP17 
                 MC3 
                 MC3/DSPC/Chol/PEG-DSG 
               
               
                   
                   
                 50/10/38.5/1.5 
               
               
                   
                   
                 Lipid:siRNA: 10:1 
               
               
                 LNP18 
                 MC3 
                 MC3/DSPC/Chol/PEG-DMG 
               
               
                   
                   
                 50/10/38.5/1.5 
               
               
                   
                   
                 Lipid:siRNA: 12:1 
               
               
                 LNP19 
                 MC3 
                 MC3/DSPC/Chol/PEG-DMG 
               
               
                   
                   
                 50/10/35/5 
               
               
                   
                   
                 Lipid:siRNA: 8:1 
               
               
                 LNP20 
                 MC3 
                 MC3/DSPC/Chol/PEG-DPG 
               
               
                   
                   
                 50/10/38.5/1.5 
               
               
                   
                   
                 Lipid:siRNA: 10:1 
               
               
                 LNP21 
                 C12-200 
                 C12-200/DSPC/Chol/PEG-DSG 
               
               
                   
                   
                 50/10/38.5/1.5 
               
               
                   
                   
                 Lipid:siRNA: 7:1 
               
               
                 LNP22 
                 XTC 
                 XTC/DSPC/Chol/PEG-DSG 
               
               
                   
                   
                 50/10/38.5/1.5 
               
               
                   
                   
                 Lipid:siRNA: 10:1 
               
               
                   
               
               
                 DSPC: distearoylphosphatidylcholine 
               
               
                 DPPC: dipalmitoylphosphatidylcholine 
               
               
                 PEG-DMG: PEG-didimyristoyl glycerol (C14-PEG, or PEG-C14) (PEG with avg mol wt of 2000) 
               
               
                 PEG-DSG: PEG-distyryl glycerol (C18-PEG, or PEG-C18) (PEG with avg mol wt of 2000) 
               
               
                 PEG-cDMA: PEG-carbamoyl-1,2-dimyristyloxypropylamine (PEG with avg mol wt of 2000) 
               
               
                 SNALP (l,2-Dilinolenyloxy-N,N-dimethylaminopropane (DLinDMA)) comprising formulations are described in WO 2009/127060, which is hereby incorporated by reference. 
               
            
           
         
       
     
     XTC comprising formulations are described in WO 2010/088537, the entire contents of which are hereby incorporated herein by reference. 
     MC3 comprising formulations are described, e.g., in United States Patent Publication No. 2010/0324120, the entire contents of which are hereby incorporated by reference. 
     ALNY-100 comprising formulations are described in WO 2010/054406, the entire contents of which are hereby incorporated herein by reference. 
     C12-200 comprising formulations are described in WO 2010/129709, the entire contents of which are hereby incorporated herein by reference. 
     Compositions and formulations for oral administration include powders or granules, microparticulates, nanoparticulates, suspensions or solutions in water or non-aqueous media, capsules, gel capsules, sachets, tablets or minitablets. Thickeners, flavoring agents, diluents, emulsifiers, dispersing aids or binders can be desirable. In some embodiments, oral formulations are those in which dsRNAs featured in the disclosure are administered in conjunction with one or more penetration enhancer surfactants and chelators. Suitable surfactants include fatty acids or esters or salts thereof, bile acids or salts thereof. Suitable bile acids/salts include chenodeoxycholic acid (CDCA) and ursodeoxychenodeoxycholic acid (UDCA), cholic acid, dehydrocholic acid, deoxycholic acid, glucholic acid, glycholic acid, glycodeoxycholic acid, taurocholic acid, taurodeoxycholic acid, sodium tauro-24,25-dihydro-fusidate and sodium glycodihydrofusidate. Suitable fatty acids include arachidonic acid, undecanoic acid, oleic acid, lauric acid, caprylic acid, capric acid, myristic acid, palmitic acid, stearic acid, linoleic acid, linolenic acid, dicaprate, tricaprate, monoolein, dilaurin, glyceryl 1-monocaprate, 1-dodecylazacycloheptan-2-one, an acylcarnitine, an acylcholine, or a monoglyceride, a diglyceride or a pharmaceutically acceptable salt thereof (e.g., sodium). In some embodiments, combinations of penetration enhancers are used, for example, fatty acids/salts in combination with bile acids/salts. One exemplary combination is the sodium salt of lauric acid, capric acid and UDCA. Further penetration enhancers include polyoxyethylene-9-lauryl ether, polyoxyethylene-20-cetyl ether. DsRNAs featured in the disclosure can be delivered orally, in granular form including sprayed dried particles, or complexed to form micro or nanoparticles. DsRNA complexing agents include poly-amino acids; polyimines; polyacrylates; polyalkylacrylates, polyoxethanes, polyalkylcyanoacrylates; cationized gelatins, albumins, starches, acrylates, polyethyleneglycols (PEG) and starches; polyalkylcyanoacrylates; DEAE-derivatized polyimines, pollulans, celluloses and starches. Suitable complexing agents include chitosan, N-trimethylchitosan, poly-L-lysine, polyhistidine, polyornithine, polyspermines, protamine, polyvinylpyridine, polythiodiethylaminomethylethylene P(TDAE), polyaminostyrene (e.g., p-amino), poly(methylcyanoacrylate), poly(ethylcyanoacrylate), poly(butylcyanoacrylate), poly(isobutylcyanoacrylate), poly(isohexylcynaoacrylate), DEAE-methacrylate, DEAE-hexylacrylate, DEAE-acrylamide, DEAE-albumin and DEAE-dextran, polymethylacrylate, polyhexylacrylate, poly(D,L-lactic acid), poly(DL-lactic-co-glycolic acid (PLGA), alginate, and polyethyleneglycol (PEG). Oral formulations for dsRNAs and their preparation are described in detail in U.S. Pat. No. 6,887,906, U.S. 2003/0027780, and U.S. Pat. No. 6,747,014, each of which is incorporated herein by reference. 
     Compositions for pulmonary system delivery may include aqueous solutions, e.g., for intranasal or oral inhalative administration, suitable carriers composed of, e.g., lipids (liposomes, niosomes, microemulsions, lipidic micelles, solid lipid nanoparticles) or polymers (polymer micelles, dendrimers, polymeric nanoparticles, nonogels, nanocapsules), adjuvant, e.g., for oral inhalative administration. Aqueous compositions may be sterile and may optionally contain buffers, diluents, absorbtion enhancers and other suitable additives. 
     Compositions and formulations for parenteral, intraparenchymal (into the brain), intrathecal, intraventricular or intrahepatic administration can include sterile aqueous solutions which can also contain buffers, diluents and other suitable additives such as, but not limited to, penetration enhancers, carrier compounds and other pharmaceutically acceptable carriers or excipients. 
     Pharmaceutical compositions of the present disclosure include, but are not limited to, solutions, emulsions, and liposome-containing formulations. These compositions can be generated from a variety of components that include, but are not limited to, preformed liquids, self-emulsifying solids and self-emulsifying semisolids. Particularly preferred are formulations that target the brain when treating APP-associated diseases or disorders. 
     The pharmaceutical formulations of the present disclosure, which can conveniently be presented in unit dosage form, can be prepared according to conventional techniques well known in the pharmaceutical industry. Such techniques include the step of bringing into association the active ingredients with the pharmaceutical carrier(s) or excipient(s). In general, the formulations are prepared by uniformly and intimately bringing into association the active ingredients with liquid carriers or finely divided solid carriers or both, and then, if necessary, shaping the product. 
     The compositions of the present disclosure can be formulated into any of many possible dosage forms such as, but not limited to, tablets, capsules, gel capsules, liquid syrups, soft gels, suppositories, and enemas. The compositions of the present disclosure can also be formulated as suspensions in aqueous, non-aqueous or mixed media. Aqueous suspensions can further contain substances which increase the viscosity of the suspension including, for example, sodium carboxymethylcellulose, sorbitol or dextran. The suspension can also contain stabilizers. 
     Additional Formulations 
     i. Emulsions 
     The compositions of the present disclosure can be prepared and formulated as emulsions. Emulsions are typically heterogeneous systems of one liquid dispersed in another in the form of droplets usually exceeding 0.1 μm in diameter (see e.g., Ansel&#39;s Pharmaceutical Dosage Forms and Drug Delivery Systems, Allen, L V., Popovich N G., and Ansel H C., 2004, Lippincott Williams &amp; Wilkins (8th ed.), New York, N.Y.; Idson, in Pharmaceutical Dosage Forms, Lieberman, Rieger and Banker (Eds.), 1988, Marcel Dekker, Inc., New York, N.Y., volume 1, p. 199; Rosoff, in Pharmaceutical Dosage Forms, Lieberman, Rieger and Banker (Eds.), 1988, Marcel Dekker, Inc., New York, N.Y., Volume 1, p. 245; Block in Pharmaceutical Dosage Forms, Lieberman, Rieger and Banker (Eds.), 1988, Marcel Dekker, Inc., New York, N.Y., volume 2, p. 335; Higuchi et al., in Remington&#39;s Pharmaceutical Sciences, Mack Publishing Co., Easton, Pa., 1985, p. 301). Emulsions are often biphasic systems comprising two immiscible liquid phases intimately mixed and dispersed with each other. In general, emulsions can be of either the water-in-oil (w/o) or the oil-in-water (o/w) variety. When an aqueous phase is finely divided into and dispersed as minute droplets into a bulk oily phase, the resulting composition is called a water-in-oil (w/o) emulsion. Alternatively, when an oily phase is finely divided into and dispersed as minute droplets into a bulk aqueous phase, the resulting composition is called an oil-in-water (o/w) emulsion. Emulsions can contain additional components in addition to the dispersed phases, and the active drug which can be present as a solution in either aqueous phase, oily phase or itself as a separate phase. Pharmaceutical excipients such as emulsifiers, stabilizers, dyes, and anti-oxidants can also be present in emulsions as needed. Pharmaceutical emulsions can also be multiple emulsions that are comprised of more than two phases such as, for example, in the case of oil-in-water-in-oil (o/w/o) and water-in-oil-in-water (w/o/w) emulsions. Such complex formulations often provide certain advantages that simple binary emulsions do not. Multiple emulsions in which individual oil droplets of an o/w emulsion enclose small water droplets constitute a w/o/w emulsion. Likewise, a system of oil droplets enclosed in globules of water stabilized in an oily continuous phase provides an o/w/o emulsion. 
     Emulsions are characterized by little or no thermodynamic stability. Often, the dispersed or discontinuous phase of the emulsion is well dispersed into the external or continuous phase and maintained in this form through the means of emulsifiers or the viscosity of the formulation. Either of the phases of the emulsion can be a semisolid or a solid, as is the case of emulsion-style ointment bases and creams Other means of stabilizing emulsions entail the use of emulsifiers that can be incorporated into either phase of the emulsion. Emulsifiers can broadly be classified into four categories: synthetic surfactants, naturally occurring emulsifiers, absorption bases, and finely dispersed solids (see e.g., Ansel&#39;s Pharmaceutical Dosage Forms and Drug Delivery Systems, Allen, L V., Popovich N G., and Ansel H C., 2004, Lippincott Williams &amp; Wilkins (8th ed.), New York, N.Y.; Idson, in Pharmaceutical Dosage Forms, Lieberman, Rieger and Banker (Eds.), 1988, Marcel Dekker, Inc., New York, N.Y., volume 1, p. 199). 
     Synthetic surfactants, also known as surface active agents, have found wide applicability in the formulation of emulsions and have been reviewed in the literature (see e.g., Ansel&#39;s Pharmaceutical Dosage Forms and Drug Delivery Systems, Allen, L V., Popovich N G., and Ansel H C., 2004, Lippincott Williams &amp; Wilkins (8th ed.), New York, N.Y.; Rieger, in Pharmaceutical Dosage Forms, Lieberman, Rieger and Banker (Eds.), 1988, Marcel Dekker, Inc., New York, N.Y., volume 1, p. 285; Idson, in Pharmaceutical Dosage Forms, Lieberman, Rieger and Banker (Eds.), Marcel Dekker, Inc., New York, N.Y., 1988, volume 1, p. 199). Surfactants are typically amphiphilic and comprise a hydrophilic and a hydrophobic portion. The ratio of the hydrophilic to the hydrophobic nature of the surfactant has been termed the hydrophile/lipophile balance (HLB) and is a valuable tool in categorizing and selecting surfactants in the preparation of formulations. Surfactants can be classified into different classes based on the nature of the hydrophilic group: nonionic, anionic, cationic and amphoteric (see e.g., Ansel&#39;s Pharmaceutical Dosage Forms and Drug Delivery Systems, Allen, L V., Popovich N G., and Ansel H C., 2004, Lippincott Williams &amp; Wilkins (8th ed.), New York, N.Y. Rieger, in Pharmaceutical Dosage Forms, Lieberman, Rieger and Banker (Eds.), 1988, Marcel Dekker, Inc., New York, N.Y., volume 1, p. 285). 
     Naturally occurring emulsifiers used in emulsion formulations include lanolin, beeswax, phosphatides, lecithin and acacia. Absorption bases possess hydrophilic properties such that they can soak up water to form w/o emulsions yet retain their semisolid consistencies, such as anhydrous lanolin and hydrophilic petrolatum. Finely divided solids have also been used as good emulsifiers especially in combination with surfactants and in viscous preparations. These include polar inorganic solids, such as heavy metal hydroxides, nonswelling clays such as bentonite, attapulgite, hectorite, kaolin, montmorillonite, colloidal aluminum silicate and colloidal magnesium aluminum silicate, pigments and nonpolar solids such as carbon or glyceryl tristearate. 
     A large variety of non-emulsifying materials are also included in emulsion formulations and contribute to the properties of emulsions. These include fats, oils, waxes, fatty acids, fatty alcohols, fatty esters, humectants, hydrophilic colloids, preservatives and antioxidants (Block, in Pharmaceutical Dosage Forms, Lieberman, Rieger and Banker (Eds.), 1988, Marcel Dekker, Inc., New York, N.Y., volume 1, p. 335; Idson, in Pharmaceutical Dosage Forms, Lieberman, Rieger and Banker (Eds.), 1988, Marcel Dekker, Inc., New York, N.Y., volume 1, p. 199). 
     Hydrophilic colloids or hydrocolloids include naturally occurring gums and synthetic polymers such as polysaccharides (for example, acacia, agar, alginic acid, carrageenan, guar gum, karaya gum, and tragacanth), cellulose derivatives (for example, carboxymethylcellulose and carboxypropylcellulose), and synthetic polymers (for example, carbomers, cellulose ethers, and carboxyvinyl polymers). These disperse or swell in water to form colloidal solutions that stabilize emulsions by forming strong interfacial films around the dispersed-phase droplets and by increasing the viscosity of the external phase. 
     Since emulsions often contain a number of ingredients such as carbohydrates, proteins, sterols and phosphatides that can readily support the growth of microbes, these formulations often incorporate preservatives. Commonly used preservatives included in emulsion formulations include methyl paraben, propyl paraben, quaternary ammonium salts, benzalkonium chloride, esters of p-hydroxybenzoic acid, and boric acid. Antioxidants are also commonly added to emulsion formulations to prevent deterioration of the formulation. Antioxidants used can be free radical scavengers such as tocopherols, alkyl gallates, butylated hydroxyanisole, butylated hydroxytoluene, or reducing agents such as ascorbic acid and sodium metabisulfite, and antioxidant synergists such as citric acid, tartaric acid, and lecithin. 
     The application of emulsion formulations via dermatological, oral and parenteral routes and methods for their manufacture have been reviewed in the literature (see e.g., Ansel&#39;s Pharmaceutical Dosage Forms and Drug Delivery Systems, Allen, L V., Popovich N G., and Ansel H C., 2004, Lippincott Williams &amp; Wilkins (8th ed.), New York, N.Y.; Idson, in Pharmaceutical Dosage Forms, Lieberman, Rieger and Banker (Eds.), 1988, Marcel Dekker, Inc., New York, N.Y., volume 1, p. 199). Emulsion formulations for oral delivery have been very widely used because of ease of formulation, as well as efficacy from an absorption and bioavailability standpoint (see e.g., Ansel&#39;s Pharmaceutical Dosage Forms and Drug Delivery Systems, Allen, L V., Popovich N G., and Ansel H C., 2004, Lippincott Williams &amp; Wilkins (8th ed.), New York, N.Y.; Rosoff, in Pharmaceutical Dosage Forms, Lieberman, Rieger and Banker (Eds.), 1988, Marcel Dekker, Inc., New York, N.Y., volume 1, p. 245; Idson, in Pharmaceutical Dosage Forms, Lieberman, Rieger and Banker (Eds.), 1988, Marcel Dekker, Inc., New York, N.Y., volume 1, p. 199). Mineral-oil base laxatives, oil-soluble vitamins and high fat nutritive preparations are among the materials that have commonly been administered orally as o/w emulsions. 
     ii. Microemulsions 
     In one embodiment of the present disclosure, the compositions of RNAi agents and nucleic acids are formulated as microemulsions. A microemulsion can be defined as a system of water, oil and amphiphile which is a single optically isotropic and thermodynamically stable liquid solution (see e.g., Ansel&#39;s Pharmaceutical Dosage Forms and Drug Delivery Systems, Allen, L V., Popovich N G., and Ansel H C., 2004, Lippincott Williams &amp; Wilkins (8th ed.), New York, N.Y.; Rosoff, in Pharmaceutical Dosage Forms, Lieberman, Rieger and Banker (Eds.), 1988, Marcel Dekker, Inc., New York, N.Y., volume 1, p. 245). Typically, microemulsions are systems that are prepared by first dispersing an oil in an aqueous surfactant solution and then adding a sufficient amount of a fourth component, generally an intermediate chain-length alcohol to form a transparent system. Therefore, microemulsions have also been described as thermodynamically stable, isotropically clear dispersions of two immiscible liquids that are stabilized by interfacial films of surface-active molecules (Leung and Shah, in: Controlled Release of Drugs: Polymers and Aggregate Systems, Rosoff, M., Ed., 1989, VCH Publishers, New York, pages 185-215). Microemulsions commonly are prepared via a combination of three to five components that include oil, water, surfactant, cosurfactant and electrolyte. Whether the microemulsion is of the water-in-oil (w/o) or an oil-in-water (o/w) type is dependent on the properties of the oil and surfactant used, and on the structure and geometric packing of the polar heads and hydrocarbon tails of the surfactant molecules (Schott, in  Remington&#39;s  Pharmaceutical Sciences, Mack Publishing Co., Easton, Pa., 1985, p. 271). 
     The phenomenological approach utilizing phase diagrams has been extensively studied and has yielded a comprehensive knowledge, to one skilled in the art, of how to formulate microemulsions (see e.g., Ansel&#39;s Pharmaceutical Dosage Forms and Drug Delivery Systems, Allen, L V., Popovich N G., and Ansel H C., 2004, Lippincott Williams &amp; Wilkins (8th ed.), New York, N.Y.; Rosoff, in Pharmaceutical Dosage Forms, Lieberman, Rieger and Banker (Eds.), 1988, Marcel Dekker, Inc., New York, N.Y., volume 1, p. 245; Block, in Pharmaceutical Dosage Forms, Lieberman, Rieger and Banker (Eds.), 1988, Marcel Dekker, Inc., New York, N.Y., volume 1, p. 335). Compared to conventional emulsions, microemulsions offer the advantage of solubilizing water-insoluble drugs in a formulation of thermodynamically stable droplets that are formed spontaneously. 
     Surfactants used in the preparation of microemulsions include, but are not limited to, ionic surfactants, non-ionic surfactants, Brij 96, polyoxyethylene oleyl ethers, polyglycerol fatty acid esters, tetraglycerol monolaurate (ML310), tetraglycerol monooleate (MO310), hexaglycerol monooleate (PO310), hexaglycerol pentaoleate (PO500), decaglycerol monocaprate (MCA750), decaglycerol monooleate (MO750), decaglycerol sequioleate (SO750), decaglycerol decaoleate (DAO750), alone or in combination with cosurfactants. The cosurfactant, usually a short-chain alcohol such as ethanol, 1-propanol, and 1-butanol, serves to increase the interfacial fluidity by penetrating into the surfactant film and consequently creating a disordered film because of the void space generated among surfactant molecules. Microemulsions can, however, be prepared without the use of cosurfactants and alcohol-free self-emulsifying microemulsion systems are known in the art. The aqueous phase can typically be, but is not limited to, water, an aqueous solution of the drug, glycerol, PEG300, PEG400, polyglycerols, propylene glycols, and derivatives of ethylene glycol. The oil phase can include, but is not limited to, materials such as Captex 300, Captex 355, Capmul MCM, fatty acid esters, medium chain (C8-C12) mono, di, and tri-glycerides, polyoxyethylated glyceryl fatty acid esters, fatty alcohols, polyglycolized glycerides, saturated polyglycolized C8-C10 glycerides, vegetable oils and silicone oil. 
     Microemulsions are particularly of interest from the standpoint of drug solubilization and the enhanced absorption of drugs. Lipid based microemulsions (both o/w and w/o) have been proposed to enhance the oral bioavailability of drugs, including peptides (see e.g., U.S. Pat. Nos. 6,191,105; 7,063,860; 7,070,802; 7,157,099; Constantinides et al., Pharmaceutical Research, 1994, 11, 1385-1390; Ritschel, Meth. Find. Exp. Clin. Pharmacol., 1993, 13, 205). Microemulsions afford advantages of improved drug solubilization, protection of drug from enzymatic hydrolysis, possible enhancement of drug absorption due to surfactant-induced alterations in membrane fluidity and permeability, ease of preparation, ease of oral administration over solid dosage forms, improved clinical potency, and decreased toxicity (see e.g., U.S. Pat. Nos. 6,191,105; 7,063,860; 7,070,802; 7,157,099; Constantinides et al., Pharmaceutical Research, 1994, 11, 1385; Ho et al., J. Pharm. Sci., 1996, 85, 138-143). Often microemulsions can form spontaneously when their components are brought together at ambient temperature. This can be particularly advantageous when formulating thermolabile drugs, peptides or RNAi agents. Microemulsions have also been effective in the transdermal delivery of active components in both cosmetic and pharmaceutical applications. It is expected that the microemulsion compositions and formulations of the present disclosure will facilitate the increased systemic absorption of RNAi agents and nucleic acids from the gastrointestinal tract, as well as improve the local cellular uptake of RNAi agents and nucleic acids. 
     Microemulsions of the present disclosure can also contain additional components and additives such as sorbitan monostearate (Grill 3), Labrasol, and penetration enhancers to improve the properties of the formulation and to enhance the absorption of the RNAi agents and nucleic acids of the present disclosure. Penetration enhancers used in the microemulsions of the present disclosure can be classified as belonging to one of five broad categories—surfactants, fatty acids, bile salts, chelating agents, and non-chelating non-surfactants (Lee et al., Critical Reviews in Therapeutic Drug Carrier Systems, 1991, p. 92). Each of these classes has been discussed above. 
     iii. Microparticles 
     An RNAi agent of the disclosure may be incorporated into a particle, e.g., a microparticle. Microparticles can be produced by spray-drying, but may also be produced by other methods including lyophilization, evaporation, fluid bed drying, vacuum drying, or a combination of these techniques. 
     iv. Penetration Enhancers 
     In one embodiment, the present disclosure employs various penetration enhancers to effect the efficient delivery of nucleic acids, particularly RNAi agents, to the skin of animals. Most drugs are present in solution in both ionized and nonionized forms. However, usually only lipid soluble or lipophilic drugs readily cross cell membranes. It has been discovered that even non-lipophilic drugs can cross cell membranes if the membrane to be crossed is treated with a penetration enhancer. In addition to aiding the diffusion of non-lipophilic drugs across cell membranes, penetration enhancers also enhance the permeability of lipophilic drugs. 
     Penetration enhancers can be classified as belonging to one of five broad categories, i.e., surfactants, fatty acids, bile salts, chelating agents, and non-chelating non-surfactants (see e.g., Malmsten, M. Surfactants and polymers in drug delivery, Informa Health Care, New York, N.Y., 2002; Lee et al., Critical Reviews in Therapeutic Drug Carrier Systems, 1991, p. 92). Each of the above mentioned classes of penetration enhancers are described below in greater detail. 
     Surfactants (or “surface-active agents”) are chemical entities which, when dissolved in an aqueous solution, reduce the surface tension of the solution or the interfacial tension between the aqueous solution and another liquid, with the result that absorption of RNAi agents through the mucosa is enhanced. In addition to bile salts and fatty acids, these penetration enhancers include, for example, sodium lauryl sulfate, polyoxyethylene-9-lauryl ether and polyoxyethylene-20-cetyl ether) (see e.g., Malmsten, M. Surfactants and polymers in drug delivery, Informa Health Care, New York, N.Y., 2002; Lee et al., Critical Reviews in Therapeutic Drug Carrier Systems, 1991, p. 92); and perfluorochemical emulsions, such as FC-43. Takahashi et al., J. Pharm. Pharmacol., 1988, 40, 252). 
     Various fatty acids and their derivatives which act as penetration enhancers include, for example, oleic acid, lauric acid, capric acid (n-decanoic acid), myristic acid, palmitic acid, stearic acid, linoleic acid, linolenic acid, dicaprate, tricaprate, monoolein (1-monooleoyl-rac-glycerol), dilaurin, caprylic acid, arachidonic acid, glycerol 1-monocaprate, 1-dodecylazacycloheptan-2-one, acylcarnitines, acylcholines, C 1-20  alkyl esters thereof (e.g., methyl, isopropyl and t-butyl), and mono- and di-glycerides thereof (i.e., oleate, laurate, caprate, myristate, palmitate, stearate, linoleate, etc.) (see e.g., Touitou, E., et al. Enhancement in Drug Delivery, CRC Press, Danvers, Mass., 2006; Lee et al., Critical Reviews in Therapeutic Drug Carrier Systems, 1991, p. 92; Muranishi, Critical Reviews in Therapeutic Drug Carrier Systems, 1990, 7, 1-33; El Hariri et al., J. Pharm. Pharmacol., 1992, 44, 651-654). 
     The physiological role of bile includes the facilitation of dispersion and absorption of lipids and fat-soluble vitamins (see e.g., Malmsten, M. Surfactants and polymers in drug delivery, Informa Health Care, New York, N.Y., 2002; Brunton, Chapter 38 in: Goodman &amp; Gilman&#39;s The Pharmacological Basis of Therapeutics, 9th Ed., Hardman et al. Eds., McGraw-Hill, New York, 1996, pp. 934-935). Various natural bile salts, and their synthetic derivatives, act as penetration enhancers. Thus the term “bile salts” includes any of the naturally occurring components of bile as well as any of their synthetic derivatives. Suitable bile salts include, for example, cholic acid (or its pharmaceutically acceptable sodium salt, sodium cholate), dehydrocholic acid (sodium dehydrocholate), deoxycholic acid (sodium deoxycholate), glucholic acid (sodium glucholate), glycholic acid (sodium glycocholate), glycodeoxycholic acid (sodium glycodeoxycholate), taurocholic acid (sodium taurocholate), taurodeoxycholic acid (sodium taurodeoxycholate), chenodeoxycholic acid (sodium chenodeoxycholate), ursodeoxycholic acid (UDCA), sodium tauro-24,25-dihydro-fusidate (STDHF), sodium glycodihydrofusidate and polyoxyethylene-9-lauryl ether (POE) (see e.g., Malmsten, M. Surfactants and polymers in drug delivery, Informa Health Care, New York, N.Y., 2002; Lee et al., Critical Reviews in Therapeutic Drug Carrier Systems, 1991, page 92; Swinyard, Chapter 39 In: Remington&#39;s Pharmaceutical Sciences, 18th Ed., Gennaro, ed., Mack Publishing Co., Easton, Pa., 1990, pages 782-783; Muranishi, Critical Reviews in Therapeutic Drug Carrier Systems, 1990, 7, 1-33; Yamamoto et al., J. Pharm. Exp. Ther., 1992, 263, 25; Yamashita et al., J. Pharm. Sci., 1990, 79, 579-583). 
     Chelating agents, as used in connection with the present disclosure, can be defined as compounds that remove metallic ions from solution by forming complexes therewith, with the result that absorption of RNAi agents through the mucosa is enhanced. With regards to their use as penetration enhancers in the present disclosure, chelating agents have the added advantage of also serving as DNase inhibitors, as most characterized DNA nucleases require a divalent metal ion for catalysis and are thus inhibited by chelating agents (Jarrett, J. Chromatogr., 1993, 618, 315-339). Suitable chelating agents include but are not limited to disodium ethylenediaminetetraacetate (EDTA), citric acid, salicylates (e.g., sodium salicylate, 5-methoxysalicylate and homovanilate), N-acyl derivatives of collagen, laureth-9 and N-amino acyl derivatives of beta-diketones (enamines)(see e.g., Katdare, A. et al., Excipient development for pharmaceutical, biotechnology, and drug delivery, CRC Press, Danvers, Mass., 2006; Lee et al., Critical Reviews in Therapeutic Drug Carrier Systems, 1991, page 92; Muranishi, Critical Reviews in Therapeutic Drug Carrier Systems, 1990, 7, 1-33; Buur et al., J. Control Rd., 1990, 14, 43-51). 
     As used herein, non-chelating non-surfactant penetration enhancing compounds can be defined as compounds that demonstrate insignificant activity as chelating agents or as surfactants but that nonetheless enhance absorption of RNAi agents through the alimentary mucosa (see e.g., Muranishi, Critical Reviews in Therapeutic Drug Carrier Systems, 1990, 7, 1-33). This class of penetration enhancers includes, for example, unsaturated cyclic ureas, 1-alkyl- and 1-alkenylazacyclo-alkanone derivatives (Lee et al., Critical Reviews in Therapeutic Drug Carrier Systems, 1991, page 92); and non-steroidal anti-inflammatory agents such as diclofenac sodium, indomethacin and phenylbutazone (Yamashita et al., J. Pharm. Pharmacol., 1987, 39, 621-626). 
     Agents that enhance uptake of RNAi agents at the cellular level can also be added to the pharmaceutical and other compositions of the present disclosure. For example, cationic lipids, such as lipofectin (Junichi et al, U.S. Pat. No. 5,705,188), cationic glycerol derivatives, and polycationic molecules, such as polylysine (WO 97/30731), are also known to enhance the cellular uptake of dsRNAs. 
     Other agents can be utilized to enhance the penetration of the administered nucleic acids, including glycols such as ethylene glycol and propylene glycol, pyrrols such as 2-pyrrol, azones, and terpenes such as limonene and menthone. 
     vi. Excipients 
     In contrast to a carrier compound, a “pharmaceutical carrier” or “excipient” is a pharmaceutically acceptable solvent, suspending agent or any other pharmacologically inert vehicle for delivering one or more nucleic acids to an animal. The excipient can be liquid or solid and is selected, with the planned manner of administration in mind, so as to provide for the desired bulk, consistency, etc., when combined with a nucleic acid and the other components of a given pharmaceutical composition. Typical pharmaceutical carriers include, but are not limited to, binding agents (e.g., pregelatinized maize starch, polyvinylpyrrolidone or hydroxypropyl methylcellulose, etc.); fillers (e.g., lactose and other sugars, microcrystalline cellulose, pectin, gelatin, calcium sulfate, ethyl cellulose, polyacrylates or calcium hydrogen phosphate, etc.); lubricants (e.g., magnesium stearate, talc, silica, colloidal silicon dioxide, stearic acid, metallic stearates, hydrogenated vegetable oils, corn starch, polyethylene glycols, sodium benzoate, sodium acetate, etc.); disintegrants (e.g., starch, sodium starch glycolate, etc.); and wetting agents (e.g., sodium lauryl sulphate, etc). 
     Pharmaceutically acceptable organic or inorganic excipients suitable for non-parenteral administration which do not deleteriously react with nucleic acids can also be used to formulate the compositions of the present disclosure. Suitable pharmaceutically acceptable carriers include, but are not limited to, water, salt solutions, alcohols, polyethylene glycols, gelatin, lactose, amylose, magnesium stearate, talc, silicic acid, viscous paraffin, hydroxymethylcellulose, polyvinylpyrrolidone and the like. 
     Formulations for topical administration of nucleic acids can include sterile and non-sterile aqueous solutions, non-aqueous solutions in common solvents such as alcohols, or solutions of the nucleic acids in liquid or solid oil bases. The solutions can also contain buffers, diluents and other suitable additives. Pharmaceutically acceptable organic or inorganic excipients suitable for non-parenteral administration which do not deleteriously react with nucleic acids can be used. 
     Suitable pharmaceutically acceptable excipients include, but are not limited to, water, salt solutions, alcohol, polyethylene glycols, gelatin, lactose, amylose, magnesium stearate, talc, silicic acid, viscous paraffin, hydroxymethylcellulose, polyvinylpyrrolidone and the like. 
     vii. Other Components 
     The compositions of the present disclosure can additionally contain other adjunct components conventionally found in pharmaceutical compositions, at their art-established usage levels. Thus, for example, the compositions can contain additional, compatible, pharmaceutically-active materials such as, for example, antipruritics, astringents, local anesthetics or anti-inflammatory agents, or can contain additional materials useful in physically formulating various dosage forms of the compositions of the present disclosure, such as dyes, flavoring agents, preservatives, antioxidants, opacifiers, thickening agents and stabilizers. However, such materials, when added, should not unduly interfere with the biological activities of the components of the compositions of the present disclosure. The formulations can be sterilized and, if desired, mixed with auxiliary agents, e.g., lubricants, preservatives, stabilizers, wetting agents, emulsifiers, salts for influencing osmotic pressure, buffers, colorings, flavorings or aromatic substances and the like which do not deleteriously interact with the nucleic acid(s) of the formulation. 
     Aqueous suspensions can contain substances which increase the viscosity of the suspension including, for example, sodium carboxymethylcellulose, sorbitol or dextran. The suspension can also contain stabilizers. 
     In some embodiments, pharmaceutical compositions featured in the disclosure include (a) one or more RNAi agents and (b) one or more agents which function by a non-RNAi mechanism and which are useful in treating an ACE2-associated disorder. Examples of such agents include, but are not limited to an antiviral agent, an immune stimulator, a therapeutic vaccine, a viral entry inhibitor, and a combination of any of the foregoing. 
     Toxicity and therapeutic efficacy of such compounds can be determined by standard pharmaceutical procedures in cell cultures or experimental animals, e.g., for determining the LD 50  (the dose lethal to 50% of the population) and the ED 50  (the dose therapeutically effective in 50% of the population). The dose ratio between toxic and therapeutic effects is the therapeutic index and it can be expressed as the ratio LD 50 /ED 50 . Compounds that exhibit high therapeutic indices are preferred. 
     The data obtained from cell culture assays and animal studies can be used in formulating a range of dosage for use in humans. The dosage of compositions featured herein in the disclosure lies generally within a range of circulating concentrations that include the ED 50  with little or no toxicity. The dosage can vary within this range depending upon the dosage form employed and the route of administration utilized. For any compound used in the methods featured in the disclosure, the therapeutically effective dose can be estimated initially from cell culture assays. A dose can be formulated in animal models to achieve a circulating plasma concentration range of the compound or, when appropriate, of the polypeptide product of a target sequence (e.g., achieving a decreased concentration of the polypeptide) that includes the IC 50  (i.e., the concentration of the test compound which achieves a half-maximal inhibition of symptoms) as determined in cell culture. Such information can be used to more accurately determine useful doses in humans. Levels in plasma can be measured, for example, by high performance liquid chromatography. 
     In addition to their administration, as discussed above, the RNAi agents featured in the disclosure can be administered in combination with other known agents effective in treatment of pathological processes mediated by nucleotide repeat expression. In any event, the administering physician can adjust the amount and timing of RNAi agent administration on the basis of results observed using standard measures of efficacy known in the art or described herein. 
     VII. KITS 
     In certain aspects, the instant disclosure provides kits that include a suitable container containing a pharmaceutical formulation of a siRNA compound, e.g., a double-stranded siRNA compound, or siRNA compound, (e.g., a precursor, e.g., a larger siRNA compound which can be processed into a siRNA compound, or a DNA which encodes an siRNA compound, e.g., a double-stranded siRNA compound, or siRNA compound, or precursor thereof). In certain embodiments the individual components of the pharmaceutical formulation may be provided in one container. Alternatively, it may be desirable to provide the components of the pharmaceutical formulation separately in two or more containers, e.g., one container for a siRNA compound preparation, and at least another for a carrier compound. The kit may be packaged in a number of different configurations such as one or more containers in a single box. The different components can be combined, e.g., according to instructions provided with the kit. The components can be combined according to a method described herein, e.g., to prepare and administer a pharmaceutical composition. The kit can also include a delivery device, such as a device suitable for pulmonary administration, e.g., a device suitable for oral inhalative administration including nebulizers, metered-dose inhalers, and dry powder inhalers. 
     VIII. METHODS FOR INHIBITING ACE2 EXPRESSION 
     The present disclosure also provides methods of inhibiting expression of an ACE2 gene in a cell. The methods include contacting a cell with an RNAi agent, e.g., double stranded RNAi agent, in an amount effective to inhibit expression of an ACE2 gene in the cell, thereby inhibiting expression of ACE2 in the cell. In certain embodiments of the disclosure, expression of an ACE2 gene is inhibited preferentially in the pulmonary system (e.g., lung, bronchial, alveoli) cells. In other embodiments of the disclosure, expression of an ACE2 gene is inhibited preferentially in the liver (e.g., hepatocytes). In certain embodiments of the disclosure, expression of an ACE2 gene is inhibited in the pulmonary system (e.g., lung, bronchial, alveoli) cells and in liver (e.g., hepatocytes) cells. 
     Contacting of a cell with a RNAi agent, e.g., a double stranded RNAi agent, may be done in vitro or in vivo. Contacting a cell in vivo with the RNAi agent includes contacting a cell or group of cells within a subject, e.g., a human subject, with the RNAi agent. Combinations of in vitro and in vivo methods of contacting a cell are also possible. 
     Contacting a cell may be direct or indirect, as discussed above. Furthermore, contacting a cell may be accomplished via a targeting ligand, including any ligand described herein or known in the art. In some embodiments, the targeting ligand is a carbohydrate moiety, e.g., a GalNAc ligand, or any other ligand that directs the RNAi agent to a site of interest. 
     The term “inhibiting,” as used herein, is used interchangeably with “reducing,” “silencing,” “downregulating,” “suppressing” and other similar terms, and includes any level of inhibition. In certain embodiments, a level of inhibition, e.g., for an RNAi agent of the instant disclosure, can be assessed in cell culture conditions, e.g., wherein cells in cell culture are transfected via Lipofectamine™-mediated transfection at a concentration in the vicinity of a cell of 10 nM or less, 1 nM or less, etc. Knockdown of a given RNAi agent can be determined via comparison of pre-treated levels in cell culture versus post-treated levels in cell culture, optionally also comparing against cells treated in parallel with a scrambled or other form of control RNAi agent. Knockdown in cell culture of, e.g., preferably 50% or more, can thereby be identified as indicative of “inhibiting” or “reducing”, “downregulating” or “suppressing”, etc. having occurred. It is expressly contemplated that assessment of targeted mRNA or encoded protein levels (and therefore an extent of “inhibiting”, etc. caused by a RNAi agent of the disclosure) can also be assessed in in vivo systems for the RNAi agents of the instant disclosure, under properly controlled conditions as described in the art. 
     The phrase “inhibiting expression of an ACE2 gene” or “inhibiting expression of ACE2,” as used herein, includes inhibition of expression of any ACE2 gene (such as, e.g., a mouse ACE2 gene, a rat ACE2 gene, a monkey ACE2 gene, or a human ACE2 gene) as well as variants or mutants of an ACE2 gene that encode an ACE2 protein. Thus, the ACE2 gene may be a wild-type ACE2 gene, a mutant ACE2 gene, or a transgenic ACE2 gene in the context of a genetically manipulated cell, group of cells, or organism. 
     “Inhibiting expression of an ACE2 gene” includes any level of inhibition of an ACE2 gene, e.g., at least partial suppression of the expression of an ACE2 gene, such as an inhibition by at least 20%. In certain embodiments, inhibition is by at least 30%, at least 40%, preferably at least 50%, at least about 60%, at least 70%, at least about 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%; or to below the level of detection of the assay method. In a preferred method, inhibition is measured at a 10 nM concentration of the siRNA using the luciferase assay provided in Example 1. 
     The expression of an ACE2 gene may be assessed based on the level of any variable associated with ACE2 gene expression, e.g., ACE2 mRNA level or ACE2 protein level. 
     Inhibition may be assessed by a decrease in an absolute or relative level of one or more of these variables compared with a control level. The control level may be any type of control level that is utilized in the art, e.g., a pre-dose baseline level, or a level determined from a similar subject, cell, or sample that is untreated or treated with a control (such as, e.g., buffer only control or inactive agent control). 
     In some embodiments of the methods of the disclosure, expression of an ACE2 gene is inhibited by at least 20%, 30%, 40%, preferably at least 50%, 60%, 70%, 80%, 85%, 90%, or 95%, or to below the level of detection of the assay. In certain embodiments, the methods include a clinically relevant inhibition of expression of ACE2, e.g. as demonstrated by a clinically relevant outcome after treatment of a subject with an agent to reduce the expression of an ACE2 gene. 
     Inhibition of the expression of an ACE2 gene may be manifested by a reduction of the amount of mRNA expressed by a first cell or group of cells (such cells may be present, for example, in a sample derived from a subject) in which an ACE2 gene is transcribed and which has or have been treated (e.g., by contacting the cell or cells with a RNAi agent of the disclosure, or by administering a RNAi agent of the disclosure to a subject in which the cells are or were present) such that the expression of an ACE2 gene is inhibited, as compared to a second cell or group of cells substantially identical to the first cell or group of cells but which has not or have not been so treated (control cell(s) not treated with a RNAi agent or not treated with a RNAi agent targeted to the genome of interest). The degree of inhibition may be expressed in terms of: 
     
       
         
           
             
               
                 
                   
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     In other embodiments, inhibition of the expression of an ACE2 gene may be assessed in terms of a reduction of a parameter that is functionally linked to an ACE2 gene expression, e.g., ACE2 protein expression, S protein priming, efficiency of viral entry, viral load. ACE2 gene silencing may be determined in any cell expressing an ACE2 gene, either endogenous or heterologous from an expression construct, and by any assay known in the art. 
     Inhibition of the expression of an ACE2 protein may be manifested by a reduction in the level of the ACE2 protein that is expressed by a cell or group of cells (e.g., the level of protein expressed in a sample derived from a subject). As explained above, for the assessment of genome suppression, the inhibition of protein expression levels in a treated cell or group of cells may similarly be expressed as a percentage of the level of protein in a control cell or group of cells. 
     A control cell or group of cells that may be used to assess the inhibition of the expression of an ACE2 gene includes a cell or group of cells that has not yet been contacted with an RNAi agent of the disclosure. For example, the control cell or group of cells may be derived from an individual subject (e.g., a human or animal subject) prior to treatment of the subject with an RNAi agent. 
     The level of ACE2 mRNA that is expressed by a cell or group of cells may be determined using any method known in the art for assessing RNA expression. In one embodiment, the level of expression of ACE2 in a sample is determined by detecting a transcribed polynucleotide, or portion thereof, e.g., mRNA of the ACE2 gene. RNA may be extracted from cells using RNA extraction techniques including, for example, using acid phenol/guanidine isothiocyanate extraction (RNAzol B; Biogenesis), RNeasy™ RNA preparation kits (Qiagen®) or PAXgene (PreAnalytix, Switzerland). Typical assay formats utilizing ribonucleic acid hybridization include nuclear run-on assays, RT-PCR, RNase protection assays, northern blotting, in situ hybridization, and microarray analysis. Circulating ACE2 mRNA may be detected using methods the described in WO2012/177906, the entire contents of which are hereby incorporated herein by reference. 
     In some embodiments, the level of expression of ACE2 is determined using a nucleic acid probe. The term “probe”, as used herein, refers to any molecule that is capable of selectively binding to a specific ACE2 nucleic acid or protein, or fragment thereof. Probes can be synthesized by one of skill in the art, or derived from appropriate biological preparations. Probes may be specifically designed to be labeled. Examples of molecules that can be utilized as probes include, but are not limited to, RNA, DNA, proteins, antibodies, and organic molecules. 
     Isolated mRNA can be used in hybridization or amplification assays that include, but are not limited to, Southern or northern analyses, polymerase chain reaction (PCR) analyses and probe arrays. One method for the determination of RNA levels involves contacting the isolated RNA with a nucleic acid molecule (probe) that can hybridize to ACE2 RNA. In one embodiment, the RNA is immobilized on a solid surface and contacted with a probe, for example by running the isolated RNA on an agarose gel and transferring the RNA from the gel to a membrane, such as nitrocellulose. In an alternative embodiment, the probe(s) are immobilized on a solid surface and the RNA is contacted with the probe(s), for example, in an Affymetrix® gene chip array. A skilled artisan can readily adapt known RNA detection methods for use in determining the level of ACE2 mRNA. 
     An alternative method for determining the level of expression of ACE2 in a sample involves the process of nucleic acid amplification or reverse transcriptase (to prepare cDNA) of for example mRNA in the sample, e.g., by RT-PCR (the experimental embodiment set forth in Mullis, 1987, U.S. Pat. No. 4,683,202), ligase chain reaction (Barany (1991)  Proc. Natl. Acad. Sci. USA  88:189-193), self sustained sequence replication (Guatelli et al. (1990)  Proc. Natl. Acad. Sci. USA  87:1874-1878), transcriptional amplification system (Kwoh et al. (1989)  Proc. Natl. Acad. Sci. USA  86:1173-1177), Q-Beta Replicase (Lizardi et al. (1988)  Bio/Technology  6:1197), rolling circle replication (Lizardi et al., U.S. Pat. No. 5,854,033) or any other nucleic acid amplification method, followed by the detection of the amplified molecules using techniques well known to those of skill in the art. These detection schemes are especially useful for the detection of nucleic acid molecules if such molecules are present in very low numbers. In particular aspects of the disclosure, the level of expression of ACE2 is determined by quantitative fluorogenic RT-PCR (i.e., the TaqMan™ System), by a Dual-Glo® Luciferase assay, or by other art-recognized method for measurement of ACE2 expression or mRNA level. 
     The expression level of ACE2 mRNA may be monitored using a membrane blot (such as used in hybridization analysis such as northern, Southern, dot, and the like), or microwells, sample tubes, gels, beads or fibers (or any solid support comprising bound nucleic acids). See U.S. Pat. Nos. 5,770,722, 5,874,219, 5,744,305, 5,677,195 and 5,445,934, which are incorporated herein by reference. The determination of ACE2 expression level may also comprise using nucleic acid probes in solution. 
     In some embodiments, the level of RNA expression is assessed using branched DNA (bDNA) assays or real time PCR (qPCR). The use of this PCR method is described and exemplified in the Examples presented herein. Such methods can also be used for the detection of ACE2 nucleic acids. 
     The level of ACE2 protein expression may be determined using any method known in the art for the measurement of protein levels. Such methods include, for example, electrophoresis, capillary electrophoresis, high performance liquid chromatography (HPLC), thin layer chromatography (TLC), hyperdiffusion chromatography, fluid or gel precipitin reactions, absorption spectroscopy, a colorimetric assays, spectrophotometric assays, flow cytometry, immunodiffusion (single or double), immunoelectrophoresis, western blotting, radioimmunoassay (RIA), enzyme-linked immunosorbent assays (ELISAs), immunofluorescent assays, electrochemiluminescence assays, and the like. Such assays can also be used for the detection of proteins indicative of the presence or replication of ACE2 proteins. 
     In some embodiments, the efficacy of the methods of the disclosure in the treatment of an ACE2-related disease is assessed by a decrease in ACE2 mRNA level (e.g, by assessment of an ACE2 level, e.g., in the lung, by biopsy, or otherwise). 
     In some embodiments, the efficacy of the methods of the disclosure in the treatment of an ACE2-related disease is assessed by a decrease in ACE2 mRNA level (e.g, by assessment of a liver sample for ACE2 level, by biopsy, or otherwise). 
     In some embodiments of the methods of the disclosure, the RNAi agent is administered to a subject such that the RNAi agent is delivered to a specific site within the subject. The inhibition of expression of ACE2 may be assessed using measurements of the level or change in the level of ACE2 mRNA or ACE2 protein in a sample derived from a specific site within the subject, e.g., lung and/or liver cells. In certain embodiments, the methods include a clinically relevant inhibition of expression of ACE2, e.g. as demonstrated by a clinically relevant outcome after treatment of a subject with an agent to reduce the expression of ACE2. 
     As used herein, the terms detecting or determining a level of an analyte are understood to mean performing the steps to determine if a material, e.g., protein, RNA, is present. As used herein, methods of detecting or determining include detection or determination of an analyte level that is below the level of detection for the method used. 
     IX. METHODS OF TREATING OR PREVENTING ACE2-ASSOCIATED DISEASES 
     The present disclosure also provides methods of using a RNAi agent of the disclosure or a composition containing a RNAi agent of the disclosure to reduce or inhibit ACE2 expression in a cell. The methods include contacting the cell with a dsRNA of the disclosure and maintaining the cell for a time sufficient to obtain degradation of the mRNA transcripts of an ACE2 gene, thereby inhibiting expression of the ACE2 gene in the cell. Reduction in gene expression can be assessed by any methods known in the art. For example, a reduction in the expression of ACE2 may be determined by determining the mRNA expression level of an ACE2 gene using methods routine to one of ordinary skill in the art, e.g., northern blotting, qRT-PCR; by determining the protein level of an ACE2 protein using methods routine to one of ordinary skill in the art, such as western blotting, immunological techniques. 
     In the methods of the disclosure the cell may be contacted in vitro or in vivo, i.e., the cell may be within a subject. 
     A cell suitable for treatment using the methods of the disclosure may be any cell that expresses an ACE2 gene. A cell suitable for use in the methods of the disclosure may be a mammalian cell, e.g., a primate cell (such as a human cell or a non-human primate cell, e.g., a monkey cell or a chimpanzee cell), a non-primate cell (such as a rat cell, or a mouse cell. In one embodiment, the cell is a human cell, e.g., a human lung cell. In one embodiment, the cell is a human cell, e.g., a human liver cell. In one embodiment, the cell is a human cell, e.g., a human lung cell and a human liver cell. 
     ACE2 expression is inhibited in the cell by at least about 30, 40, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 96, 97, 98, 99, or about 100%, i.e., to below the level of detection. In preferred embodiments, ACE2 expression is inhibited by at least 50%. 
     The in vivo methods of the disclosure may include administering to a subject a composition containing a RNAi agent, where the RNAi agent includes a nucleotide sequence that is complementary to at least a part of an RNA transcript of the ACE2 gene of the subject to be treated. When the organism to be treated is a mammal such as a human, the composition can be administered by any means known in the art including, but not limited to oral, intraperitoneal, or parenteral routes, including intracranial (e.g., intraventricular, intraparenchymal, and intrathecal), intravenous, intramuscular, subcutaneous, transdermal, airway (aerosol), nasal, rectal, and topical (including buccal and sublingual) administration. In certain embodiments, the compositions are administered by intravenous infusion or injection. In certain embodiments, the compositions are administered by subcutaneous injection. In certain embodiments, the compositions are administered by pulmonary delivery, e.g., oral inhalation or intranasal delivery. 
     In some embodiments, the administration is via a depot injection. A depot injection may release the RNAi agent in a consistent way over a prolonged time period. Thus, a depot injection may reduce the frequency of dosing needed to obtain a desired effect, e.g., a desired inhibition of ACE2, or a therapeutic or prophylactic effect. A depot injection may also provide more consistent serum concentrations. Depot injections may include subcutaneous injections or intramuscular injections. In preferred embodiments, the depot injection is a subcutaneous injection. 
     In one embodiment, the double-stranded RNAi agent is administered by pulmonary system administration, e.g., intranasal administration or oral inhalative administration. Pulmonary system administration may be via a syringe, a dropper, atomization, or use of device, e.g., a passive breath driven or active power driven single/-multiple dose dry powder inhaler (DPI) device. 
     The mode of administration may be chosen based upon whether local or systemic treatment is desired and based upon the area to be treated. The route and site of administration may be chosen to enhance targeting. 
     In one aspect, the present disclosure also provides methods for inhibiting the expression of an ACE2 gene in a mammal. The methods include administering to the mammal a composition comprising a dsRNA that targets an ACE2 gene in a cell of the mammal and maintaining the mammal for a time sufficient to obtain degradation of the RNA transcript of the ACE2 gene, thereby inhibiting expression of the ACE2 gene in the cell. Reduction in genome expression can be assessed by any methods known it the art and by methods, e.g. qRT-PCR, described herein. Reduction in protein production can be assessed by any methods known it the art and by methods, e.g. ELISA, described herein. In one embodiment, a lung biopsy sample serves as the tissue material for monitoring the reduction in ACE2 gene or protein expression (or of a proxy therefore). 
     The present disclosure further provides methods of treatment of a subject in need thereof. The treatment methods of the disclosure include administering an RNAi agent of the disclosure to a subject, e.g., a subject that would benefit from inhibition of ACE2 expression, in a therapeutically effective amount of a RNAi agent targeting an ACE2 gene or a pharmaceutical composition comprising a RNAi agent targeting an ACE2 gene. 
     In addition, the present disclosure provides methods of preventing, treating or inhibiting the progression of an ACE2-associated disease or disorder, e.g., a coronavirus-associated disease, such as severe acute respiratory syndrome (SARS), the Middle East respiratory syndrome (MERS), and severe acute respiratory syndrome-2 (SARS-2). 
     The methods include administering to the subject a therapeutically effective amount of any of the RNAi agent, e.g., dsRNA agents, or the pharmaceutical composition provided herein, thereby preventing, treating, or inhibiting the progression of the ACE2-associated disease or disorder in the subject, such as COVID-19. 
     An RNAi agent of the disclosure may be administered as a “free RNAi agent.” A free RNAi agent is administered in the absence of a pharmaceutical composition. The naked RNAi agent may be in a suitable buffer solution. The buffer solution may comprise acetate, citrate, prolamine, carbonate, or phosphate, or any combination thereof. In one embodiment, the buffer solution is phosphate buffered saline (PBS). The pH and osmolarity of the buffer solution containing the RNAi agent can be adjusted such that it is suitable for administering to a subject. 
     Alternatively, an RNAi agent of the disclosure may be administered as a pharmaceutical composition, such as a dsRNA liposomal formulation. 
     Subjects that would benefit from a reduction or inhibition of ACE2 gene expression are those having an ACE2-associated disease, subjects at risk of developing an ACE2-associate disease, e.g., subjects of an age greater than 60 years and/or subjects who are immunocompromised, and subjects at risk of developing an ACE2-associate disease, e.g., during an epidemic or pandemic. 
     The disclosure further provides methods for the use of a RNAi agent or a pharmaceutical composition thereof, e.g., for treating a subject that would benefit from reduction or inhibition of ACE2 expression, e.g., a subject having an ACE2-associated disorder, in combination with other pharmaceuticals or other therapeutic methods, e.g., with known pharmaceuticals or known therapeutic methods, such as, for example, those which are currently employed for treating these disorders. For example, in certain embodiments, an RNAi agent targeting ACE2 is administered in combination with, e.g., an agent useful in treating an ACE2-associated disorder as described elsewhere herein or as otherwise known in the art. For example, additional agents and treatments suitable for treating a subject that would benefit from reduction in ACE2 expression, e.g., a subject having an ACE2-associated disorder, may include agents currently used to treat symptoms of ACE2-associated disorders. The RNAi agent and additional therapeutic agents may be administered at the same time or in the same combination, e.g., via pulmonary system administration, or the additional therapeutic agent can be administered as part of a separate composition or at separate times or by another method known in the art or described herein. 
     Exemplary additional therapeutics and treatments include, for example, an antiviral agent, an immune stimulator, a therapeutic vaccine, a viral entry inhibitor, and a combination of any of the foregoing. 
     In one embodiment, the method includes administering a composition featured herein such that expression of the target ACE2 gene is decreased, for at least one month. In preferred embodiments, expression is decreased for at least 2 months, or 6 months. 
     Preferably, the RNAi agents useful for the methods and compositions featured herein specifically target RNAs (primary or processed) of the target ACE2 gene. Compositions and methods for inhibiting the expression of these genes using RNAi agents can be prepared and performed as described herein. 
     Administration of the dsRNA according to the methods of the disclosure may result in a reduction of the severity, signs, symptoms, or markers of such diseases or disorders in a patient with an ACE2-associated disorder. By “reduction” in this context is meant a statistically significant or clinically significant decrease in such level. The reduction can be, for example, at least 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or about 100%. 
     Efficacy of treatment or prevention of disease can be assessed, for example by measuring disease progression, disease remission, symptom severity, reduction in pain, quality of life, dose of a medication required to sustain a treatment effect, level of a disease marker or any other measurable parameter appropriate for a given disease being treated or targeted for prevention. It is well within the ability of one skilled in the art to monitor efficacy of treatment or prevention by measuring any one of such parameters, or any combination of parameters. It is well within the ability of one skilled in the art to monitor efficacy of treatment or prevention by measuring any one of such parameters, or any combination of parameters. In connection with the administration of a RNAi agent targeting ACE2 or pharmaceutical composition thereof, “effective against” an ACE2-associated disorder indicates that administration in a clinically appropriate manner results in a beneficial effect for at least a statistically significant fraction of patients, such as an improvement of symptoms, a cure, a reduction in disease, extension of life, improvement in quality of life, or other effect generally recognized as positive by medical doctors familiar with treating ACE2-associated disorders and the related causes. 
     A treatment or preventive effect is evident when there is a statistically significant improvement in one or more parameters of disease status, or by a failure to worsen or to develop symptoms where they would otherwise be anticipated. As an example, a favorable change of at least 10% in a measurable parameter of disease, and preferably at least 20%, 30%, 40%, 50%, or more can be indicative of effective treatment. Efficacy for a given RNAi agent drug or formulation of that drug can also be judged using an experimental animal model for the given disease as known in the art. When using an experimental animal model, efficacy of treatment is evidenced when a statistically significant reduction in a marker or symptom is observed. 
     Alternatively, the efficacy can be measured by a reduction in the severity of disease as determined by one skilled in the art of diagnosis based on a clinically accepted disease severity grading scale. Any positive change resulting in e.g., lessening of severity of disease measured using the appropriate scale, represents adequate treatment using a RNAi agent or RNAi agent formulation as described herein. 
     Subjects can be administered a therapeutic amount of dsRNA, such as about 0.01 mg/kg to about 200 mg/kg. 
     The RNAi agent can be administered via the pulmonary system over a period of time, on a regular basis. In certain embodiments, after an initial treatment regimen, the treatments can be administered on a less frequent basis. Administration of the RNAi agent can reduce ACE2 levels, e.g., in a cell, tissue, blood, lung sample or other compartment of the patient by at least 20%, 30%, 40%, 50%, 55%, 60%, 65%, 70,% 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, or at least about 99% or more. In a preferred embodiment, administration of the RNAi agent can reduce ACE2 levels, e.g., in a cell, tissue, blood, pulmonary system sample or other compartment of the patient by at least 50%. 
     Before administration of a full dose of the RNAi agent, patients can be administered a smaller dose, such as a 5% infusion reaction, and monitored for adverse effects, such as an allergic reaction. In another example, the patient can be monitored for unwanted immunostimulatory effects, such as increased cytokine (e.g., TNF-alpha or INF-alpha) levels. 
     Alternatively, the RNAi agent can be administered by pulmonary administration or subcutaneously, i.e., by subcutaneous injection. One or more injections may be used to deliver the desired, e.g., monthly dose of RNAi agent to a subject. The injections may be repeated over a period of time. The administration may be repeated on a regular basis. In certain embodiments, after an initial treatment regimen, the treatments can be administered on a less frequent basis. A repeat-dose regimen may include administration of a therapeutic amount of RNAi agent on a regular basis, such as monthly or extending to once a quarter, twice per year, once per year. In certain embodiments, the RNAi agent is administered about once per month to about once per quarter (i.e., about once every three months). 
     Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. Although methods and materials similar or equivalent to those described herein can be used in the practice or testing of the RNAi agents and methods featured in the invention, suitable methods and materials are described below. All publications, patent applications, patents, and other references mentioned herein are incorporated by reference in their entirety. In case of conflict, the present specification, including definitions, will control. In addition, the materials, methods, and examples are illustrative only and not intended to be limiting. 
     An informal Sequence Listing is filed herewith and forms part of the specification as filed. 
     This invention is further illustrated by the following examples which should not be construed as limiting. The entire contents of all references, patents and published patent applications cited throughout this application, as well as the Figures and the Sequence Listing, are hereby incorporated herein by reference. 
     EXAMPLES 
     Example 1. iRNA Synthesis 
     Source of Reagents 
     Where the source of a reagent is not specifically given herein, such reagent can be obtained from any supplier of reagents for molecular biology at a quality/purity standard for application in molecular biology. 
     siRNA Design 
     The selection of siRNA designs targeting human angiotensin converting enzyme 2 (ACE2) gene (human NCBI refseqID: NM_021804.3; NCBI GeneID: 59272) or cynomolgus monkey ACE2 gene (NCBI refseqID: XM_005593037.2) were designed using custo R and Python scripts. The human NM_021804.3 mRNA has a length of 3596 bases. The cynomolgus monkey XM_005593037.2 mRNA has a length of 3575 bases. 
     A detailed list of a set of the unmodified siRNA sense and antisense strand sequences targeting human ACE2 is shown in Table 2. 
     A detailed list of a set of the modified siRNA sense and antisense strand sequences targeting human ACE2 is shown in Table 3. 
     A detailed list of a set of the unmodified siRNA sense and antisense strand sequences targeting cynomolgus monkey ACE2 is shown in Table 4. 
     A detailed list of a set of the modified siRNA sense and antisense strand sequences targeting cynomolgus monkey ACE2 is shown in Table 5. 
     It is to be understood that, throughout the application, a duplex name without a decimal is equivalent to a duplex name with a decimal which merely references the batch number of the duplex. For example, AD-1230521 is equivalent to AD-1230521. 
     siRNA Synthesis 
     siRNAs were synthesized and annealed using routine methods known in the art. Briefly, siRNA sequences were synthesized on a 1 μmol scale using a Mermade 192 synthesizer (BioAutomation) with phosphoramidite chemistry on solid supports. The solid support was controlled pore glass (500-1000 Å) loaded with a custom GalNAc ligand (3′-GalNAc conjugates), universal solid support (AM Chemicals), or the first nucleotide of interest. Ancillary synthesis reagents and standard 2-cyanoethyl phosphoramidite monomers (2′-deoxy-2′-fluoro, 2′-O-methyl, RNA, DNA) were obtained from Thermo-Fisher (Milwaukee, Wis.), Hongene (China), or Chemgenes (Wilmington, Mass., USA). Additional phosphoramidite monomers were procured from commercial suppliers, prepared in-house, or procured using custom synthesis from various CMOs. Phosphoramidites were prepared at a concentration of 100 mM in either acetonitrile or 9:1 acetonitrile:DMF and were coupled using 5-Ethylthio-1H-tetrazole (ETT, 0.25 M in acetonitrile) with a reaction time of 400 s. Phosphorothioate linkages were generated using a 100 mM solution of 3-((Dimethylamino-methylidene) amino)-3H-1,2,4-dithiazole-3-thione (DDTT, obtained from Chemgenes (Wilmington, Mass., USA)) in anhydrous acetonitrile/pyridine (9:1 v/v). Oxidation time was 5 minutes. All sequences were synthesized with final removal of the DMT group (“DMT-Off”). 
     Upon completion of the solid phase synthesis, solid-supported oligoribonucleotides were treated with 300 μL of Methylamine (40% aqueous) at room temperature in 96 well plates for approximately 2 hours to afford cleavage from the solid support and subsequent removal of all additional base-labile protecting groups. For sequences containing any natural ribonucleotide linkages (2′-OH) protected with a tert-butyl dimethyl silyl (TBDMS) group, a second deprotection step was performed using TEA.3HF (triethylamine trihydrofluoride). To each oligonucleotide solution in aqueous methylamine was added 200 μL of dimethyl sulfoxide (DMSO) and 300 μL TEA.3HF and the solution was incubated for approximately 30 mins at 60° C. After incubation, the plate was allowed to come to room temperature and crude oligonucleotides were precipitated by the addition of 1 mL of 9:1 acetontrile:ethanol or 1:1 ethanol:isopropanol. The plates were then centrifuged at 4° C. for 45 mins and the supernatant carefully decanted with the aid of a multichannel pipette. The oligonucleotide pellet was resuspended in 20 mM NaOAc and subsequently desalted using a HiTrap size exclusion column (5 mL, GE Healthcare) on an Agilent LC system equipped with an autosampler, UV detector, conductivity meter, and fraction collector. Desalted samples were collected in 96 well plates and then analyzed by LC-MS and UV spectrometry to confirm identity and quantify the amount of material, respectively. 
     Duplexing of single strands was performed on a Tecan liquid hand ling robot. Sense and antisense single strands were combined in an equimolar ratio to a final concentration of 10 μM in 1× PBS in 96 well plates, the plate sealed, incubated at 100° C. for 10 minutes, and subsequently allowed to return slowly to room temperature over a period of 2-3 hours. The concentration and identity of each duplex was confirmed and then subsequently utilized for in vitro screening assays. 
     Example 2. In Vitro Screening of siRNA Duplexes 
     Cell Culture and Transfections 
     Cells, e.g., pulmonary system cells, are cultured according to standard methods and are transfected with the iRNA duplex of interest. For example, primary human hepatocytes (PHH) were transfected by adding 7.5 μL of Opti-MEM plus 0.1 μL of RNAiMAX per well (Invitrogen, Carlsbad Calif. cat #13778-150) to 2.5 μL of each siRNA duplex to an individual well in a 384-well plate. The cells were then incubated at room temperature for 15 minutes. Forty μL of MEDIA containing ˜1.5×10 4  cells was then added to the siRNA mixture. Cells were incubated for 24 hours prior to RNA purification. Single dose experiments were performed at 10 nM, 1 nM, and 0.1 nM. 
     Total RNA Isolation Using DYNABEADS mRNA Isolation Kit 
     Total RNA isolation was performed using DYNABEADS. Briefly, cells were lysed in 10□l of Lysis/Binding Buffer containing 3 μL of beads per well and mixed for 10 minutes on an electrostatic shaker. The washing steps were automated on a Biotek EL406, using a magnetic plate support. Beads were washed (in 3 μL) once in Buffer A, once in Buffer B, and twice in Buffer E, with aspiration steps in between. Following a final aspiration, complete 12 μL RT mixture was added to each well, as described below. 
     cDNA Synthesis 
     For cDNA synthesis, a master mix of 1.5 μl 10× Buffer, 0.6 μl 10× dNTPs, 1.5 μl Random primers, 0.75 μl Reverse Transcriptase, 0.75 μl RNase inhibitor and 9.9 μl of H2O per reaction were added per well. Plates were sealed, agitated for 10 minutes on an electrostatic shaker, and then incubated at 37 degrees C. for 2 hours. Following this, the plates were agitated at 80 degrees C. for 8 minutes. 
     Real Time PCR 
     Two microlitre (μ1) of cDNA were added to a master mix containing 0.5 μl of human GAPDH TaqMan Probe (4326317E), 0.5 μl human APOC3, 2 μl nuclease-free water and 5 μl Lightcycler 480 probe master mix (Roche Cat #04887301001) per well in a 384 well plates (Roche cat #04887301001). Real time PCR was done in a LightCycler480 Real Time PCR system (Roche). 
     To calculate relative fold change, data were analyzed using the ΔΔCt method and normalized to assays performed with cells transfected with 10 nM AD-1955, or mock transfected cells. IC50s were calculated using a 4 parameter fit model using XLFit and normalized to cells transfected with AD-1955 or mock-transfected. The sense and antisense sequences of AD-1955 are: sense: cuuAcGcuGAGuAcuucGAdTsdT (SEQ ID NO:17) and antisense UCGAAGuACUcAGCGuAAGdTsdT (SEQ ID NO:18). 
     The results of the screening of the dsRNA agents listed in Tables 3 and 5 in primary human hepatocytes (PHH) are shown in Table 6. 
     
       
         
           
               
             
               
                 TABLE 1 
               
             
            
               
                   
               
               
                 Abbreviations of nucleotide monomers used in nucleic acid sequence representation. It will 
               
               
                 be understood that these monomers, when present in an oligonucleotide, are mutually linked by 5′-3′-phosphodiester bonds. 
               
            
           
           
               
               
            
               
                 Abbreviation 
                 Nucleotide(s) 
               
               
                   
               
               
                 A 
                 Adenosine-3′-phosphate 
               
               
                 Ab 
                 beta-L-adenosine-3′-phosphate 
               
               
                 Abs 
                 beta-L-adenosine-3′-phosphorothioate 
               
               
                 Af 
                 2′-fluoroadenosine-3′-phosphate 
               
               
                 Afs 
                 2′-fluoroadenosine-3′-phosphorothioate 
               
               
                 As 
                 adenosine-3′-phosphorothioate 
               
               
                 C 
                 cytidine-3′-phosphate 
               
               
                 Cb 
                 beta-L-cytidine-3′-phosphate 
               
               
                 Cbs 
                 beta-L-cytidine-3′-phosphorothioate 
               
               
                 Cf 
                 2′-fluorocytidine-3′-phosphate 
               
               
                 Cfs 
                 2′-fluorocytidine-3′-phosphorothioate 
               
               
                 Cs 
                 cytidine-3′-phosphorothioate 
               
               
                 G 
                 guanosine-3′-phosphate 
               
               
                 Gb 
                 beta-L-guanosine-3′-phosphate 
               
               
                 Gbs 
                 beta-L-guanosine-3′-phosphorothioate 
               
               
                 Gf 
                 2′-fluoroguanosine-3′-phosphate 
               
               
                 Gfs 
                 2′-fluoroguanosine-3′-phosphorothioate 
               
               
                 Gs 
                 guanosine-3′-phosphorothioate 
               
               
                 T 
                 5′-methyluridine-3′-phosphate 
               
               
                 Tf 
                 2′-fluoro-5-methyluridine-3′-phosphate 
               
               
                 Tfs 
                 2′-fluoro-5-methyluridine-3′-phosphorothioate 
               
               
                 Ts 
                 5-methyluridine-3′-phosphorothioate 
               
               
                 U 
                 Uridine-3′-phosphate 
               
               
                 Uf 
                 2′-fluorouridine-3′-phosphate 
               
               
                 Ufs 
                 2′-fluorouridine-3′-phosphorothioate 
               
               
                 Us 
                 uridine-3′-phosphorothioate 
               
               
                 N 
                 any nucleotide, modified or unmodified 
               
               
                 a 
                 2′-O-methyladenosine-3′-phosphate 
               
               
                 as 
                 2′-O-methyladenosine-3′-phosphorothioate 
               
               
                 c 
                 2′-O-methylcytidine-3′-phosphate 
               
               
                 cs 
                 2′-O-methylcytidine-3′-phosphorothioate 
               
               
                 g 
                 2′-O-methylguanosine-3′-phosphate 
               
               
                 gs 
                 2′-O-methylguanosine-3′-phosphorothioate 
               
               
                 t 
                 2′-O-methyl-5-methyluridine-3′-phosphate 
               
               
                 ts 
                 2′-O-methyl-5-methyluridine-3′-phosphorothioate 
               
               
                 u 
                 2′-O-methyluridine-3′-phosphate 
               
               
                 us 
                 2′-O-methyluridine-3′-phosphorothioate 
               
               
                 s 
                 phosphorothioate linkage 
               
               
                 L10 
                 N-(cholesterylcarboxamidocaproyl)-4-hydroxyprolinol (Hyp-C6-Chol) 
               
               
                 L96 
                 N-[tris(GalNAc-alkyl)-amidodecanoyl)]-4-hydroxyprolinol 
               
               
                   
                 (Hyp-(GalNAc-alkyl)3) 
               
               
                   
                 
                   
                     
                     
                         
                         
                     
                   
                 
               
               
                   
               
               
                 Y34 
                 2-hydroxymethyl-tetrahydrofurane-4-methoxy-3-phosphate (abasic 2′-OMe furanose) 
               
               
                 Y44 
                 inverted abasic DNA (2-hydroxymethyl-tetrahydrofurane-5-phosphate) 
               
               
                 (Agn) 
                 Adenosine-glycol nucleic acid (GNA) 
               
               
                 (Cgn) 
                 Cytidine-glycol nucleic acid (GNA) 
               
               
                 (Ggn) 
                 Guanosine-glycol nucleic acid (GNA) 
               
               
                 (Tgn) 
                 Thymidine-glycol nucleic acid (GNA) S-Isomer 
               
               
                 P 
                 Phosphate 
               
               
                 VP 
                 Vinyl-phosphonate 
               
               
                 dA 
                 2′-deoxyadenosine-3′-phosphate 
               
               
                 dAs 
                 2′-deoxyadenosine-3′-phosphorothioate 
               
               
                 dC 
                 2′-deoxycytidine-3′-phosphate 
               
               
                 dCs 
                 2′-deoxycytidine-3′-phosphorothioate 
               
               
                 dG 
                 2′-deoxyguanosine-3′-phosphate 
               
               
                 dGs 
                 2′-deoxyguanosine-3′-phosphorothioate 
               
               
                 dT 
                 2′-deoxythymidine-3′-phosphate 
               
               
                 dTs 
                 2′-deoxythymidine-3′-phosphorothioate 
               
               
                 dU 
                 2′-deoxyuridine 
               
               
                 dUs 
                 2′-deoxyuridine-3′-phosphorothioate 
               
               
                 (C2p) 
                 cytidine-2′-phosphate 
               
               
                 (G2p) 
                 guanosine-2′-phosphate 
               
               
                 (U2p) 
                 uridine-2′-phosphate 
               
               
                 (A2p) 
                 adenosine-2′-phosphate 
               
               
                 (Chd) 
                 2′-O-hexadecyl-cytidine-3′-phosphate 
               
               
                 (Ahd) 
                 2′-O-hexadecyl-adenosine-3′-phosphate 
               
               
                 (Ghd) 
                 2′-O-hexadecyl-guanosine-3′-phosphate 
               
               
                 (Uhd) 
                 2′-O-hexadecyl-uridine-3′-phosphate 
               
               
                   
               
            
           
         
       
     
     
       
         
           
               
             
               
                 TABLE 2 
               
             
            
               
                   
               
               
                 Unmodified Sense and Antisense Strand Human ACE2 dsRNA Sequences 
               
            
           
           
               
               
               
               
               
               
               
            
               
                   
                   
                   
                   
                   
                   
                 Range in 
               
               
                   
                   
                 SEQ ID 
                 Range in 
                   
                 SEQ ID 
                 NM_021804.3 
               
               
                 Duplex Name 
                 Sense Sequence 5′ to 3′ 
                 NO: 
                 NM_021804.3 
                 Antisense Sequence 5′ to 3′ 
                 NO: 
               
               
                   
               
            
           
           
               
               
               
               
               
               
               
            
               
                 AD-1230821 
                 UAUCAAAGUUCACUUGCUUCA 
                 19 
                 303-323 
                 UGAAGCAAGUGAACUUUGAUAGA 
                 201 
                 301-323 
               
               
                   
               
               
                 AD-1230822 
                 CUGUUCUAUCAAAGUUCACUA 
                 20 
                 297-317 
                 UAGUGAACUUUGAUAGAACAGGU 
                 202 
                 295-317 
               
               
                   
               
               
                 AD-1230823 
                 UCUUCCUAAUAUGACUCAAGA 
                 21 
                 1139-1159 
                 UCUUGAGUCAUAUUAGGAAGACC 
                 203 
                 1137-1159 
               
               
                   
               
               
                 AD-1230825 
                 UUACUCAUUCAUUCGAUAUUA 
                 22 
                 1709-1729 
                 UAAUAUCGAAUGAAUGAGUAAUC 
                 204 
                 1707-1729 
               
               
                   
               
               
                 AD-1230826 
                 ACUCAUUCAUUCGAUAUUACA 
                 23 
                 1711-1731 
                 UGUAAUAUCGAAUGAAUGAGUAA 
                 205 
                 1709-1731 
               
               
                   
               
               
                 AD-1230827 
                 AAUGUGACAUCUCAAACUCUA 
                 24 
                 1804-1824 
                 UAGAGUUUGAGAUGUCACAUUUG 
                 206 
                 1802-1824 
               
               
                   
               
               
                 AD-1230828 
                 AUGUGACAUCUCAAACUCUAA 
                 25 
                 1805-1825 
                 UUAGAGUUUGAGAUGUCACAUUU 
                 207 
                 1803-1825 
               
               
                   
               
               
                 AD-1230829 
                 UGAAACCAAGAAUCUCCUUUA 
                 26 
                 2206-2226 
                 UAAAGGAGAUUCUUGGUUUCAAA 
                 208 
                 2204-2226 
               
               
                   
               
               
                 AD-1230831 
                 UUGGACAAAUCUGUACUCUUA 
                 27 
                 1004-1024 
                 UAAGAGUACAGAUUUGUCCAAAA 
                 209 
                 1002-1024 
               
               
                   
               
               
                 AD-1230832 
                 AAAUCUGUACUCUUUGACAGA 
                 28 
                 1010-1030 
                 UCUGUCAAAGAGUACAGAUUUGU 
                 210 
                 1008-1030 
               
               
                   
               
               
                 AD-1230833 
                 UUCUUUGUAUCUGUUGGUCUA 
                 29 
                 1122-1142 
                 UAGACCAACAGAUACAAAGAACU 
                 211 
                 1120-1142 
               
               
                   
               
               
                 AD-1230836 
                 UCUCUGUUCCAUGUUUCUAAA 
                 30 
                 1686-1706 
                 UUUAGAAACAUGGAACAGAGAUG 
                 212 
                 1684-1706 
               
               
                   
               
               
                 AD-1230837 
                 UCUGUUCCAUGUUUCUAAUGA 
                 31 
                 1688-1708 
                 UCAUUAGAAACAUGGAACAGAGA 
                 213 
                 1686-1708 
               
               
                   
               
               
                 AD-1230838 
                 UUCCAUGUUUCUAAUGAUUAA 
                 32 
                 1692-1712 
                 UUAAUCAUUAGAAACAUGGAACA 
                 214 
                 1690-1712 
               
               
                   
               
               
                 AD-1230839 
                 UCCAUGUUUCUAAUGAUUACA 
                 33 
                 1693-1713 
                 UGUAAUCAUUAGAAACAUGGAAC 
                 215 
                 1691-1713 
               
               
                   
               
               
                 AD-1230841 
                 UAAUGAUUACUCAUUCAUUCA 
                 34 
                 1703-1723 
                 UGAAUGAAUGAGUAAUCAUUAGA 
                 216 
                 1701-1723 
               
               
                   
               
               
                 AD-1230842 
                 GAUUACUCAUUCAUUCGAUAA 
                 35 
                 1707-1727 
                 UUAUCGAAUGAAUGAGUAAUCAU 
                 217 
                 1705-1727 
               
               
                   
               
               
                 AD-1230843 
                 CUCAUUCAUUCGAUAUUACAA 
                 36 
                 1712-1732 
                 UUGUAAUAUCGAAUGAAUGAGUA 
                 218 
                 1710-1732 
               
               
                   
               
               
                 AD-1230844 
                 GACCUGUUCUAUCAAAGUUCA 
                 37 
                 294-314 
                 UGAACUUUGAUAGAACAGGUCUU 
                 219 
                 292-314 
               
               
                   
               
               
                 AD-1230845 
                 CCUUUACCAAUUCCAGUUUCA 
                 38 
                 1739-1759 
                 UGAAACUGGAAUUGGUAAAGGGU 
                 220 
                 1737-1759 
               
               
                   
               
               
                 AD-1230846 
                 CCUGUUCUAUCAAAGUUCACA 
                 39 
                 296-316 
                 UGUGAACUUUGAUAGAACAGGUC 
                 221 
                 294-316 
               
               
                   
               
               
                 AD-1230847 
                 UGUUCUAUCAAAGUUCACUUA 
                 40 
                 298-318 
                 UAAGUGAACUUUGAUAGAACAGG 
                 222 
                 296-318 
               
               
                   
               
               
                 AD-1230848 
                 GUUCUAUCAAAGUUCACUUGA 
                 41 
                 299-319 
                 UCAAGUGAACUUUGAUAGAACAG 
                 223 
                 297-319 
               
               
                   
               
               
                 AD-1230849 
                 UCUAUCAAAGUUCACUUGCUA 
                 42 
                 301-321 
                 UAGCAAGUGAACUUUGAUAGAAC 
                 224 
                 299-321 
               
               
                   
               
               
                 AD-1230850 
                 AAAUGUGACAUCUCAAACUCA 
                 43 
                 1803-1823 
                 UGAGUUUGAGAUGUCACAUUUGU 
                 225 
                 1801-1823 
               
               
                   
               
               
                 AD-1230851 
                 CUAUCAAAGUUCACUUGCUUA 
                 44 
                 302-322 
                 UAAGCAAGUGAACUUUGAUAGAA 
                 226 
                 300-322 
               
               
                   
               
               
                 AD-1230852 
                 UGUGACAUCUCAAACUCUACA 
                 45 
                 1806-1826 
                 UGUAGAGUUUGAGAUGUCACAUU 
                 227 
                 1804-1826 
               
               
                   
               
               
                 AD-1230853 
                 CAUCUCAAACUCUACAGAAGA 
                 46 
                 1811-1831 
                 UCUUCUGUAGAGUUUGAGAUGUC 
                 228 
                 1809-1831 
               
               
                   
               
               
                 AD-1230855 
                 AUCAAAGUUCACUUGCUUCUA 
                 47 
                 304-324 
                 UAGAAGCAAGUGAACUUUGAUAG 
                 229 
                 302-324 
               
               
                   
               
               
                 AD-1230856 
                 UCAAAGUUCACUUGCUUCUUA 
                 48 
                 305-325 
                 UAAGAAGCAAGUGAACUUUGAUA 
                 230 
                 303-325 
               
               
                   
               
               
                 AD-1230857 
                 AACCAAGAAUCUCCUUUAAUA 
                 49 
                 2209-2229 
                 UAUUAAAGGAGAUUCUUGGUUUC 
                 231 
                 2207-2229 
               
               
                   
               
               
                 AD-1230858 
                 ACCAAGAAUCUCCUUUAAUUA 
                 50 
                 2210-2230 
                 UAAUUAAAGGAGAUUCUUGGUUU 
                 232 
                 2208-2230 
               
               
                   
               
               
                 AD-1230859 
                 UCAUUCCUAGAACUGAAGUUA 
                 51 
                 2263-2283 
                 UAACUUCAGUUCUAGGAAUGAUA 
                 233 
                 2261-2283 
               
               
                   
               
               
                 AD-1230867 
                 CGGUUGAACACAAUUCUAAAA 
                 52 
                 525-545 
                 UUUUAGAAUUGUGUUCAACCGUU 
                 234 
                 523-545 
               
               
                   
               
               
                 AD-1230868 
                 GGUUGAACACAAUUCUAAAUA 
                 53 
                 526-546 
                 UAUUUAGAAUUGUGUUCAACCGU 
                 235 
                 524-546 
               
               
                   
               
               
                 AD-1230869 
                 GUUGAACACAAUUCUAAAUAA 
                 54 
                 527-547 
                 UUAUUUAGAAUUGUGUUCAACCG 
                 236 
                 525-547 
               
               
                   
               
               
                 AD-1230870 
                 UGGACAAAUCUGUACUCUUUA 
                 55 
                 1005-1025 
                 UAAAGAGUACAGAUUUGUCCAAA 
                 237 
                 1003-1025 
               
               
                   
               
               
                 AD-1230871 
                 CUUCCUAAUAUGACUCAAGGA 
                 56 
                 1140-1160 
                 UCCUUGAGUCAUAUUAGGAAGAC 
                 238 
                 1138-1160 
               
               
                   
               
               
                 AD-1230874 
                 AACCUUUUCUGCUAAGAAAUA 
                 57 
                 1345-1365 
                 UAUUUCUUAGCAGAAAAGGUUGU 
                 239 
                 1343-1365 
               
               
                   
               
               
                 AD-1230877 
                 CUGUUCCAUGUUUCUAAUGAA 
                 58 
                 1689-1709 
                 UUCAUUAGAAACAUGGAACAGAG 
                 240 
                 1687-1709 
               
               
                   
               
               
                 AD-1230878 
                 UGUUCCAUGUUUCUAAUGAUA 
                 59 
                 1690-1710 
                 UAUCAUUAGAAACAUGGAACAGA 
                 241 
                 1688-1710 
               
               
                   
               
               
                 AD-1230879 
                 GUUCCAUGUUUCUAAUGAUUA 
                 60 
                 1691-1711 
                 UAAUCAUUAGAAACAUGGAACAG 
                 242 
                 1689-1711 
               
               
                   
               
               
                 AD-1230880 
                 UUCUAAUGAUUACUCAUUCAA 
                 61 
                 1700-1720 
                 UUGAAUGAGUAAUCAUUAGAAAC 
                 243 
                 1698-1720 
               
               
                   
               
               
                 AD-1230881 
                 AAUGAUUACUCAUUCAUUCGA 
                 62 
                 1704-1724 
                 UCGAAUGAAUGAGUAAUCAUUAG 
                 244 
                 1702-1724 
               
               
                   
               
               
                 AD-1230882 
                 AUUACUCAUUCAUUCGAUAUA 
                 63 
                 1708-1728 
                 UAUAUCGAAUGAAUGAGUAAUCA 
                 245 
                 1706-1728 
               
               
                   
               
               
                 AD-1230883 
                 UCAUUCAUUCGAUAUUACACA 
                 64 
                 1713-1733 
                 UGUGUAAUAUCGAAUGAAUGAGU 
                 246 
                 1711-1733 
               
               
                   
               
               
                 AD-1230884 
                 CAUUCAUUCGAUAUUACACAA 
                 65 
                 1714-1734 
                 UUGUGUAAUAUCGAAUGAAUGAG 
                 247 
                 1712-1734 
               
               
                   
               
               
                 AD-1230885 
                 AGACCUGUUCUAUCAAAGUUA 
                 66 
                 293-313 
                 UAACUUUGAUAGAACAGGUCUUC 
                 248 
                 291-313 
               
               
                   
               
               
                 AD-1230886 
                 ACCUGUUCUAUCAAAGUUCAA 
                 67 
                 295-315 
                 UUGAACUUUGAUAGAACAGGUCU 
                 249 
                 293-315 
               
               
                   
               
               
                 AD-1230887 
                 CUUUACCAAUUCCAGUUUCAA 
                 68 
                 1740-1760 
                 UUGAAACUGGAAUUGGUAAAGGG 
                 250 
                 1738-1760 
               
               
                   
               
               
                 AD-1230888 
                 UUACCAAUUCCAGUUUCAAGA 
                 69 
                 1742-1762 
                 UCUUGAAACUGGAAUUGGUAAAG 
                 251 
                 1740-1762 
               
               
                   
               
               
                 AD-1230889 
                 UUCAAGAAGCACUUUGUCAAA 
                 70 
                 1756-1776 
                 UUUGACAAAGUGCUUCUUGAAAC 
                 252 
                 1754-1776 
               
               
                   
               
               
                 AD-1230890 
                 CAAAUGUGACAUCUCAAACUA 
                 71 
                 1802-1822 
                 UAGUUUGAGAUGUCACAUUUGUG 
                 253 
                 1800-1822 
               
               
                   
               
               
                 AD-1230891 
                 UGACAUCUCAAACUCUACAGA 
                 72 
                 1808-1828 
                 UCUGUAGAGUUUGAGAUGUCACA 
                 254 
                 1806-1828 
               
               
                   
               
               
                 AD-1230892 
                 GACAUCUCAAACUCUACAGAA 
                 73 
                 1809-1829 
                 UUCUGUAGAGUUUGAGAUGUCAC 
                 255 
                 1807-1829 
               
               
                   
               
               
                 AD-1230894 
                 CAAAGUUCACUUGCUUCUUGA 
                 74 
                 306-326 
                 UCAAGAAGCAAGUGAACUUUGAU 
                 256 
                 304-326 
               
               
                   
               
               
                 AD-1230895 
                 AAAGUUCACUUGCUUCUUGGA 
                 75 
                 307-327 
                 UCCAAGAAGCAAGUGAACUUUGA 
                 257 
                 305-327 
               
               
                   
               
               
                 AD-1230896 
                 GUUCACUUGCUUCUUGGAAUA 
                 76 
                 310-330 
                 UAUUCCAAGAAGCAAGUGAACUU 
                 258 
                 308-330 
               
               
                   
               
               
                 AD-1230902 
                 UUUGACUUCUGUUCUGUUUCA 
                 79 
                 2782-2802 
                 UGAAACAGAACAGAAGUCAAAUC 
                 261 
                 2780-2802 
               
               
                   
               
               
                 AD-1230904 
                 CAAGAAAUUCAGAAUCUCACA 
                 80 
                 2782-2802 
                 UGUGAGAUUCUGAAUUUCUUGUA 
                 262 
                 436-458 
               
               
                   
               
               
                 AD-1230910 
                 GGACAAAUCUGUACUCUUUGA 
                 81 
                 1006-1026 
                 UCAAAGAGUACAGAUUUGUCCAA 
                 263 
                 1004-1026 
               
               
                   
               
               
                 AD-1230911 
                 AAUCUGUACUCUUUGACAGUA 
                 82 
                 1011-1031 
                 UACUGUCAAAGAGUACAGAUUUG 
                 264 
                 1009-1031 
               
               
                   
               
               
                 AD-1230912 
                 UCUGUACUCUUUGACAGUUCA 
                 83 
                 1013-1033 
                 UGAACUGUCAAAGAGUACAGAUU 
                 265 
                 1011-1033 
               
               
                   
               
               
                 AD-1230913 
                 UCUUUGUAUCUGUUGGUCUUA 
                 84 
                 1123-1143 
                 UAAGACCAACAGAUACAAAGAAC 
                 266 
                 1121-1143 
               
               
                   
               
               
                 AD-1230914 
                 UUCCUAAUAUGACUCAAGGAA 
                 85 
                 1141-1161 
                 UUCCUUGAGUCAUAUUAGGAAGA 
                 267 
                 1139-1161 
               
               
                   
               
               
                 AD-1230915 
                 UCCUAAUAUGACUCAAGGAUA 
                 86 
                 1142-1162 
                 UAUCCUUGAGUCAUAUUAGGAAG 
                 268 
                 1140-1162 
               
               
                   
               
               
                 AD-1230916 
                 AAGGAUUCUGGGAAAAUUCCA 
                 87 
                 1156-1176 
                 UGGAAUUUUCCCAGAAUCCUUGA 
                 269 
                 1154-1176 
               
               
                   
               
               
                 AD-1230920 
                 CAUUUAAAAUCCAUUGGUCUA 
                 88 
                 1431-1451 
                 UAGACCAAUGGAUUUUAAAUGCU 
                 270 
                 1429-1451 
               
               
                   
               
               
                 AD-1230921 
                 AAAAUCCAUUGGUCUUCUGUA 
                 89 
                 1436-1456 
                 UACAGAAGACCAAUGGAUUUUAA 
                 271 
                 1434-1456 
               
               
                   
               
               
                 AD-1230922 
                 AAAUCCAUUGGUCUUCUGUCA 
                 90 
                 1437-1457 
                 UGACAGAAGACCAAUGGAUUUUA 
                 272 
                 1435-1457 
               
               
                   
               
               
                 AD-1230923 
                 AAUCCAUUGGUCUUCUGUCAA 
                 91 
                 1438-1458 
                 UUGACAGAAGACCAAUGGAUUUU 
                 273 
                 1436-1458 
               
               
                   
               
               
                 AD-1230928 
                 GCCAUUUACUUACAUGUUAGA 
                 92 
                 1532-1552 
                 UCUAACAUGUAAGUAAAUGGCAG 
                 274 
                 1530-1552 
               
               
                   
               
               
                 AD-1230930 
                 AUCUCUGUUCCAUGUUUCUAA 
                 93 
                 1685-1705 
                 UUAGAAACAUGGAACAGAGAUGC 
                 275 
                 1683-1705 
               
               
                   
               
               
                 AD-1230931 
                 CUCUGUUCCAUGUUUCUAAUA 
                 94 
                 1687-1707 
                 UAUUAGAAACAUGGAACAGAGAU 
                 276 
                 1685-1707 
               
               
                   
               
               
                 AD-1230932 
                 CCAUGUUUCUAAUGAUUACUA 
                 95 
                 1694-1714 
                 UAGUAAUCAUUAGAAACAUGGAA 
                 277 
                 1692-1714 
               
               
                   
               
               
                 AD-1230934 
                 UGAUUACUCAUUCAUUCGAUA 
                 96 
                 1706-1726 
                 UAUCGAAUGAAUGAGUAAUCAUU 
                 278 
                 1704-1726 
               
               
                   
               
               
                 AD-1230935 
                 UUUACCAAUUCCAGUUUCAAA 
                 97 
                 1741-1761 
                 UUUGAAACUGGAAUUGGUAAAGG 
                 279 
                 1739-1761 
               
               
                   
               
               
                 AD-1230936 
                 GCACAAAUGUGACAUCUCAAA 
                 98 
                 1799-1819 
                 UUUGAGAUGUCACAUUUGUGCAG 
                 280 
                 1797-1819 
               
               
                   
               
               
                 AD-1230937 
                 AGUUCACUUGCUUCUUGGAAA 
                 99 
                 309-329 
                 UUUCCAAGAAGCAAGUGAACUUU 
                 281 
                 307-329 
               
               
                   
               
               
                 AD-1230938 
                 UGCUUCUUGGAAUUAUAACAA 
                 100 
                 317-337 
                 UUGUUAUAAUUCCAAGAAGCAAG 
                 282 
                 315-337 
               
               
                   
               
               
                 AD-1230939 
                 UGGAAUUAUAACACCAAUAUA 
                 101 
                 324-344 
                 UAUAUUGGUGUUAUAAUUCCAAG 
                 283 
                 322-344 
               
               
                   
               
               
                 AD-1230940 
                 UUGAAACCAAGAAUCUCCUUA 
                 102 
                 2205-2225 
                 UAAGGAGAUUCUUGGUUUCAAAU 
                 284 
                 2203-2225 
               
               
                   
               
               
                 AD-1230941 
                 AAACCAAGAAUCUCCUUUAAA 
                 103 
                 2208-2228 
                 UUUAAAGGAGAUUCUUGGUUUCA 
                 285 
                 2206-2228 
               
               
                   
               
               
                 AD-1230942 
                 UAUCAUUCCUAGAACUGAAGA 
                 104 
                 2261-2281 
                 UCUUCAGUUCUAGGAAUGAUAUC 
                 286 
                 2259-2281 
               
               
                   
               
               
                 AD-1230946 
                 AUCCAGGAUUCCAAAACACUA 
                 105 
                 2557-2577 
                 UAGUGUUUUGGAAUCCUGGAUUA 
                 287 
                 2555-2577 
               
               
                   
               
               
                 AD-1230947 
                 ACCUCCUUUUAGAAAAAUCUA 
                 106 
                 2589-2609 
                 UAGAUUUUUCUAAAAGGAGGUCU 
                 288 
                 2587-2609 
               
               
                   
               
               
                 AD-1230948 
                 CCUCCUUUUAGAAAAAUCUAA 
                 107 
                 2590-2610 
                 UUAGAUUUUUCUAAAAGGAGGUC 
                 289 
                 2588-2610 
               
               
                   
               
               
                 AD-1230949 
                 AUCUUCAUUGACAUUGCUUUA 
                 108 
                 2739-2759 
                 UAAAGCAAUGUCAAUGAAGAUGC 
                 290 
                 2737-2759 
               
               
                   
               
               
                 AD-1230950 
                 UCUUCAUUGACAUUGCUUUCA 
                 109 
                 2740-2760 
                 UGAAAGCAAUGUCAAUGAAGAUG 
                 291 
                 2738-2760 
               
               
                   
               
               
                 AD-1230951 
                 AGUAUUUAUUUCUGUCUCUGA 
                 110 
                 2760-2780 
                 UCAGAGACAGAAAUAAAUACUGA 
                 292 
                 2758-2780 
               
               
                   
               
               
                 AD-1230962 
                 ACAAGAAAUUCAGAAUCUCAA 
                 111 
                 437-457 
                 UUGAGAUUCUGAAUUUCUUGUAG 
                 293 
                 435-457 
               
               
                   
               
               
                 AD-1230969 
                 ACGGUUGAACACAAUUCUAAA 
                 112 
                 524-544 
                 UUUAGAAUUGUGUUCAACCGUUU 
                 294 
                 522-544 
               
               
                   
               
               
                 AD-1230970 
                 UUGAACACAAUUCUAAAUACA 
                 113 
                 528-548 
                 UGUAUUUAGAAUUGUGUUCAACC 
                 295 
                 526-548 
               
               
                   
               
               
                 AD-1230971 
                 UGAACACAAUUCUAAAUACAA 
                 114 
                 529-549 
                 UUGUAUUUAGAAUUGUGUUCAAC 
                 296 
                 527-549 
               
               
                   
               
               
                 AD-1230972 
                 UCUACAGUACUGGAAAAGUUA 
                 115 
                 559-579 
                 UAACUUUUCCAGUACUGUAGAUG 
                 297 
                 557-579 
               
               
                   
               
               
                 AD-1230973 
                 CUACAGUACUGGAAAAGUUUA 
                 116 
                 560-580 
                 UAAACUUUUCCAGUACUGUAGAU 
                 298 
                 558-580 
               
               
                   
               
               
                 AD-1230977 
                 UUUUGGACAAAUCUGUACUCA 
                 117 
                 1002-1022 
                 UGAGUACAGAUUUGUCCAAAAUC 
                 299 
                 1000-1022 
               
               
                   
               
               
                 AD-1230978 
                 UUUGGACAAAUCUGUACUCUA 
                 118 
                 1003-1023 
                 UAGAGUACAGAUUUGUCCAAAAU 
                 300 
                 1001-1023 
               
               
                   
               
               
                 AD-1230979 
                 GACAAAUCUGUACUCUUUGAA 
                 119 
                 1007-1027 
                 UUCAAAGAGUACAGAUUUGUCCA 
                 301 
                 1005-1027 
               
               
                   
               
               
                 AD-1230980 
                 CUGUACUCUUUGACAGUUCCA 
                 120 
                 1014-1034 
                 UGGAACUGUCAAAGAGUACAGAU 
                 302 
                 1012-1034 
               
               
                   
               
               
                 AD-1230981 
                 UACUCUUUGACAGUUCCCUUA 
                 121 
                 1017-1037 
                 UAAGGGAACUGUCAAAGAGUACA 
                 303 
                 1015-1037 
               
               
                   
               
               
                 AD-1230982 
                 CUCUUUGACAGUUCCCUUUGA 
                 122 
                 1019-1039 
                 UCAAAGGGAACUGUCAAAGAGUA 
                 304 
                 1017-1039 
               
               
                   
               
               
                 AD-1230983 
                 GUUCUUUGUAUCUGUUGGUCA 
                 123 
                 1121-1141 
                 UGACCAACAGAUACAAAGAACUU 
                 305 
                 1119-1141 
               
               
                   
               
               
                 AD-1230984 
                 UUGUAUCUGUUGGUCUUCCUA 
                 124 
                 1126-1146 
                 UAGGAAGACCAACAGAUACAAAG 
                 306 
                 1124-1146 
               
               
                   
               
               
                 AD-1230986 
                 CUAAUAUGACUCAAGGAUUCA 
                 125 
                 1144-1164 
                 UGAAUCCUUGAGUCAUAUUAGGA 
                 307 
                 1142-1164 
               
               
                   
               
               
                 AD-1230987 
                 UAAUAUGACUCAAGGAUUCUA 
                 126 
                 1145-1165 
                 UAGAAUCCUUGAGUCAUAUUAGG 
                 308 
                 1143-1165 
               
               
                   
               
               
                 AD-1230988 
                 UCAAGGAUUCUGGGAAAAUUA 
                 127 
                 1154-1174 
                 UAAUUUUCCCAGAAUCCUUGAGU 
                 309 
                 1152-1174 
               
               
                   
               
               
                 AD-1230989 
                 CAAGGAUUCUGGGAAAAUUCA 
                 128 
                 1155-1175 
                 UGAAUUUUCCCAGAAUCCUUGAG 
                 310 
                 1153-1175 
               
               
                   
               
               
                 AD-1230990 
                 AGGAUUCUGGGAAAAUUCCAA 
                 129 
                 1157-1177 
                 UUGGAAUUUUCCCAGAAUCCUUG 
                 311 
                 1155-1177 
               
               
                   
               
               
                 AD-1230992 
                 GGGAAAUCAUGUCACUUUCUA 
                 130 
                 1396-1416 
                 UAGAAAGUGACAUGAUUUCCCCA 
                 312 
                 1394-1416 
               
               
                   
               
               
                 AD-1230993 
                 UAAAAUCCAUUGGUCUUCUGA 
                 131 
                 1435-1455 
                 UCAGAAGACCAAUGGAUUUUAAA 
                 313 
                 1433-1455 
               
               
                   
               
               
                 AD-1230994 
                 AUCCAUUGGUCUUCUGUCACA 
                 132 
                 1439-1459 
                 UGUGACAGAAGACCAAUGGAUUU 
                 314 
                 1437-1459 
               
               
                   
               
               
                 AD-1230996 
                 AUGUUUCUAAUGAUUACUCAA 
                 133 
                 1696-1716 
                 UUGAGUAAUCAUUAGAAACAUGG 
                 315 
                 1694-1716 
               
               
                   
               
               
                 AD-1230997 
                 CAGUUUCAAGAAGCACUUUGA 
                 134 
                 1752-1772 
                 UCAAAGUGCUUCUUGAAACUGGA 
                 316 
                 1750-1772 
               
               
                   
               
               
                 AD-1230998 
                 CUGCACAAAUGUGACAUCUCA 
                 135 
                 1797-1817 
                 UGAGAUGUCACAUUUGUGCAGAG 
                 317 
                 1795-1817 
               
               
                   
               
               
                 AD-1230999 
                 CACUUGCUUCUUGGAAUUAUA 
                 136 
                 313-333 
                 UAUAAUUCCAAGAAGCAAGUGAA 
                 318 
                 311-333 
               
               
                   
               
               
                 AD-1231001 
                 UUUGAAACCAAGAAUCUCCUA 
                 137 
                 2204-2224 
                 UAGGAGAUUCUUGGUUUCAAAUU 
                 319 
                 2202-2224 
               
               
                   
               
               
                 AD-1231002 
                 AUCAUUCCUAGAACUGAAGUA 
                 138 
                 2262-2282 
                 UACUUCAGUUCUAGGAAUGAUAU 
                 320 
                 2260-2282 
               
               
                   
               
               
                 AD-1231003 
                 CAUUCCUAGAACUGAAGUUGA 
                 139 
                 2264-2284 
                 UCAACUUCAGUUCUAGGAAUGAU 
                 321 
                 2262-2284 
               
               
                   
               
               
                 AD-1231004 
                 UUCCUAGAACUGAAGUUGAAA 
                 140 
                 2266-2286 
                 UUUCAACUUCAGUUCUAGGAAUG 
                 322 
                 2264-2286 
               
               
                   
               
               
                 AD-1231009 
                 UUCAUUGACAUUGCUUUCAGA 
                 141 
                 2742-2762 
                 UCUGAAAGCAAUGUCAAUGAAGA 
                 323 
                 2740-2762 
               
               
                   
               
               
                 AD-1231010 
                 CAUUGACAUUGCUUUCAGUAA 
                 142 
                 2744-2764 
                 UUACUGAAAGCAAUGUCAAUGAA 
                 324 
                 2742-2764 
               
               
                   
               
               
                 AD-1231011 
                 ACAUUGCUUUCAGUAUUUAUA 
                 143 
                 2749-2769 
                 UAUAAAUACUGAAAGCAAUGUCA 
                 325 
                 2747-2769 
               
               
                   
               
               
                 AD-1231012 
                 CAGUAUUUAUUUCUGUCUCUA 
                 144 
                 2759-2779 
                 UAGAGACAGAAAUAAAUACUGAA 
                 326 
                 2757-2779 
               
               
                   
               
               
                 AD-1231017 
                 GGAUUUGACUUCUGUUCUGUA 
                 145 
                 2779-2799 
                 UACAGAACAGAAGUCAAAUCCAG 
                 327 
                 2777-2799 
               
               
                   
               
               
                 AD-1231022 
                 UACAAGAAAUUCAGAAUCUCA 
                 146 
                 436-456 
                 UGAGAUUCUGAAUUUCUUGUAGU 
                 328 
                 434-456 
               
               
                   
               
               
                 AD-1231033 
                 CAAACGGUUGAACACAAUUCA 
                 147 
                 521-541 
                 UGAAUUGUGUUCAACCGUUUGCU 
                 329 
                 519-541 
               
               
                   
               
               
                 AD-1231034 
                 AACGGUUGAACACAAUUCUAA 
                 148 
                 523-543 
                 UUAGAAUUGUGUUCAACCGUUUG 
                 330 
                 521-543 
               
               
                   
               
               
                 AD-1231036 
                 AUCUACAGUACUGGAAAAGUA 
                 149 
                 558-578 
                 UACUUUUCCAGUACUGUAGAUGG 
                 331 
                 556-578 
               
               
                   
               
               
                 AD-1231037 
                 UACAGUACUGGAAAAGUUUGA 
                 150 
                 561-581 
                 UCAAACUUUUCCAGUACUGUAGA 
                 332 
                 559-581 
               
               
                   
               
               
                 AD-1231038 
                 ACAGUACUGGAAAAGUUUGUA 
                 151 
                 562-582 
                 UACAAACUUUUCCAGUACUGUAG 
                 333 
                 560-582 
               
               
                   
               
               
                 AD-1231039 
                 UACUGGAAAAGUUUGUAACCA 
                 152 
                 566-586 
                 UGGUUACAAACUUUUCCAGUACU 
                 334 
                 564-586 
               
               
                   
               
               
                 AD-1231043 
                 CAUUAUAUGAACAUCUUCAUA 
                 153 
                 886-906 
                 UAUGAAGAUGUUCAUAUAAUGGU 
                 335 
                 884-906 
               
               
                   
               
               
                 AD-1231044 
                 AUUUUGGACAAAUCUGUACUA 
                 154 
                 1001-1021 
                 UAGUACAGAUUUGUCCAAAAUCU 
                 336 
                 999-1021 
               
               
                   
               
               
                 AD-1231045 
                 ACAAAUCUGUACUCUUUGACA 
                 155 
                 1008-1028 
                 UGUCAAAGAGUACAGAUUUGUCC 
                 337 
                 1006-1028 
               
               
                   
               
               
                 AD-1231046 
                 CAAAUCUGUACUCUUUGACAA 
                 156 
                 1009-1029 
                 UUGUCAAAGAGUACAGAUUUGUC 
                 338 
                 1007-1029 
               
               
                   
               
               
                 AD-1231047 
                 UAUCUGUUGGUCUUCCUAAUA 
                 157 
                 1129-1149 
                 UAUUAGGAAGACCAACAGAUACA 
                 339 
                 1127-1149 
               
               
                   
               
               
                 AD-1231048 
                 UUGGUCUUCCUAAUAUGACUA 
                 158 
                 1135-1155 
                 UAGUCAUAUUAGGAAGACCAACA 
                 340 
                 1133-1155 
               
               
                   
               
               
                 AD-1231049 
                 CCUAAUAUGACUCAAGGAUUA 
                 159 
                 1143-1163 
                 UAAUCCUUGAGUCAUAUUAGGAA 
                 341 
                 1141-1163 
               
               
                   
               
               
                 AD-1231050 
                 AAUAUGACUCAAGGAUUCUGA 
                 160 
                 1146-1166 
                 UCAGAAUCCUUGAGUCAUAUUAG 
                 342 
                 1144-1166 
               
               
                   
               
               
                 AD-1231051 
                 ACCUUUUCUGCUAAGAAAUGA 
                 161 
                 1346-1366 
                 UCAUUUCUUAGCAGAAAAGGUUG 
                 343 
                 1344-1366 
               
               
                   
               
               
                 AD-1231052 
                 AAGACAUUUUUGGACAAGUUA 
                 162 
                 258-278 
                 UAACUUGUCCAAAAAUGUCUUGG 
                 344 
                 256-278 
               
               
                   
               
               
                 AD-1231053 
                 GGAAAUCAUGUCACUUUCUGA 
                 163 
                 1397-1417 
                 UCAGAAAGUGACAUGAUUUCCCC 
                 345 
                 1395-1417 
               
               
                   
               
               
                 AD-1231054 
                 AAAUCAUGUCACUUUCUGCAA 
                 164 
                 1399-1419 
                 UUGCAGAAAGUGACAUGAUUUCC 
                 346 
                 1397-1419 
               
               
                   
               
               
                 AD-1231055 
                 AAUCAUGUCACUUUCUGCAGA 
                 165 
                 1400-1420 
                 UCUGCAGAAAGUGACAUGAUUUC 
                 347 
                 1398-1420 
               
               
                   
               
               
                 AD-1231058 
                 GCAUUUAAAAUCCAUUGGUCA 
                 166 
                 1430-1450 
                 UGACCAAUGGAUUUUAAAUGCUU 
                 348 
                 1428-1450 
               
               
                   
               
               
                 AD-1231061 
                 UGAAACAGAAAUAAACUUCCA 
                 167 
                 1478-1498 
                 UGGAAGUUUAUUUCUGUUUCAUU 
                 349 
                 1476-1498 
               
               
                   
               
               
                 AD-1231065 
                 CAUCUCUGUUCCAUGUUUCUA 
                 168 
                 1684-1704 
                 UAGAAACAUGGAACAGAGAUGCG 
                 350 
                 1682-1704 
               
               
                   
               
               
                 AD-1231066 
                 UGUUUCUAAUGAUUACUCAUA 
                 169 
                 1697-1717 
                 UAUGAGUAAUCAUUAGAAACAUG 
                 351 
                 1695-1717 
               
               
                   
               
               
                 AD-1231067 
                 AUGAUUACUCAUUCAUUCGAA 
                 170 
                 1705-1725 
                 UUCGAAUGAAUGAGUAAUCAUUA 
                 352 
                 1703-1725 
               
               
                   
               
               
                 AD-1231068 
                 CAAUUCCAGUUUCAAGAAGCA 
                 171 
                 1746-1766 
                 UGCUUCUUGAAACUGGAAUUGGU 
                 353 
                 1744-1766 
               
               
                   
               
               
                 AD-1231069 
                 GUUUCAAGAAGCACUUUGUCA 
                 172 
                 1754-1774 
                 UGACAAAGUGCUUCUUGAAACUG 
                 354 
                 1752-1774 
               
               
                   
               
               
                 AD-1231070 
                 UUUCAAGAAGCACUUUGUCAA 
                 173 
                 1755-1775 
                 UUGACAAAGUGCUUCUUGAAACU 
                 355 
                 1753-1775 
               
               
                   
               
               
                 AD-1231071 
                 CUUUGUCAAGCAGCUAAACAA 
                 174 
                 1767-1787 
                 UUGUUUAGCUGCUUGACAAAGUG 
                 356 
                 1765-1787 
               
               
                   
               
               
                 AD-1231072 
                 UUCUAUCAAAGUUCACUUGCA 
                 175 
                 300-320 
                 UGCAAGUGAACUUUGAUAGAACA 
                 357 
                 298-320 
               
               
                   
               
               
                 AD-1231073 
                 AAGUUCACUUGCUUCUUGGAA 
                 176 
                 308-328 
                 UUCCAAGAAGCAAGUGAACUUUG 
                 358 
                 306-328 
               
               
                   
               
               
                 AD-1231075 
                 UUCACUUGCUUCUUGGAAUUA 
                 177 
                 311-331 
                 UAAUUCCAAGAAGCAAGUGAACU 
                 359 
                 309-331 
               
               
                   
               
               
                 AD-1231076 
                 ACUUGCUUCUUGGAAUUAUAA 
                 178 
                 314-334 
                 UUAUAAUUCCAAGAAGCAAGUGA 
                 360 
                 312-334 
               
               
                   
               
               
                 AD-1231077 
                 GCUUCUUGGAAUUAUAACACA 
                 179 
                 318-338 
                 UGUGUUAUAAUUCCAAGAAGCAA 
                 361 
                 316-338 
               
               
                   
               
               
                 AD-1231079 
                 CAGAACAAGAAUUCUUUUGUA 
                 180 
                 1974-1994 
                 UACAAAAGAAUUCUUGUUCUGGU 
                 362 
                 1972-1994 
               
               
                   
               
               
                 AD-1231080 
                 CUUCUUGGAAUUAUAACACCA 
                 181 
                 319-339 
                 UGGUGUUAUAAUUCCAAGAAGCA 
                 363 
                 317-339 
               
               
                   
               
               
                 AD-1231081 
                 CAAAGCAUCAAAGUGAGGAUA 
                 182 
                 2028-2048 
                 UAUCCUCACUUUGAUGCUUUGGU 
                 364 
                 2026-2048 
               
               
                   
               
               
                 AD-1231082 
                 AGCUCUUGGAGAUAAAGCAUA 
                 183 
                 2060-2080 
                 UAUGCUUUAUCUCCAAGAGCUGA 
                 365 
                 2058-2080 
               
               
                   
               
               
                 AD-1231083 
                 AUGUACCUGUUCCGAUCAUCA 
                 184 
                 2100-2120 
                 UGAUGAUCGGAACAGGUACAUUU 
                 366 
                 2098-2120 
               
               
                   
               
               
                 AD-1231085 
                 UUACUGAAGAGAAUGUCCAAA 
                 185 
                 343-363 
                 UUUGGACAUUCUCUUCAGUAAUA 
                 367 
                 341-363 
               
               
                   
               
               
                 AD-1231087 
                 AUUCCUAGAACUGAAGUUGAA 
                 186 
                 2265-2285 
                 UUCAACUUCAGUUCUAGGAAUGA 
                 368 
                 2263-2285 
               
               
                   
               
               
                 AD-1231090 
                 AAUCCAGGAUUCCAAAACACA 
                 187 
                 2556-2576 
                 UGUGUUUUGGAAUCCUGGAUUAU 
                 369 
                 2554-2578 
               
               
                   
               
               
                 AD-1231091 
                 UCCAGGAUUCCAAAACACUGA 
                 188 
                 2558-2578 
                 UCAGUGUUUUGGAAUCCUGGAUU 
                 370 
                 2556-2578 
               
               
                   
               
               
                 AD-1231092 
                 CCAAAACACUGAUGAUGUUCA 
                 189 
                 2567-2587 
                 UGAACAUCAUCAGUGUUUUGGAA 
                 371 
                 2565-2587 
               
               
                   
               
               
                 AD-1231093 
                 GACCUCCUUUUAGAAAAAUCA 
                 190 
                 2588-2608 
                 UGAUUUUUCUAAAAGGAGGUCUG 
                 372 
                 2586-2608 
               
               
                   
               
               
                 AD-1231097 
                 GCAUCUUCAUUGACAUUGCUA 
                 191 
                 2737-2757 
                 UAGCAAUGUCAAUGAAGAUGCUC 
                 373 
                 2735-2757 
               
               
                   
               
               
                 AD-1231098 
                 UCAUUGACAUUGCUUUCAGUA 
                 192 
                 2743-2763 
                 UACUGAAAGCAAUGUCAAUGAAG 
                 374 
                 2741-2763 
               
               
                   
               
               
                 AD-1231099 
                 AUUGACAUUGCUUUCAGUAUA 
                 193 
                 2745-2765 
                 UAUACUGAAAGCAAUGUCAAUGA 
                 375 
                 2743-2765 
               
               
                   
               
               
                 AD-1231100 
                 CAUUGCUUUCAGUAUUUAUUA 
                 194 
                 2750-2770 
                 UAAUAAAUACUGAAAGCAAUGUC 
                 376 
                 2748-2770 
               
               
                   
               
               
                 AD-1231101 
                 UUGCUUUCAGUAUUUAUUUCA 
                 195 
                 2752-2772 
                 UGAAAUAAAUACUGAAAGCAAUG 
                 377 
                 2750-2772 
               
               
                   
               
               
                 AD-1231102 
                 CUUUCAGUAUUUAUUUCUGUA 
                 196 
                 2755-2775 
                 UACAGAAAUAAAUACUGAAAGCA 
                 378 
                 2753-2775 
               
               
                   
               
               
                 AD-1231103 
                 UUUCAGUAUUUAUUUCUGUCA 
                 197 
                 2756-2776 
                 UGACAGAAAUAAAUACUGAAAGC 
                 379 
                 2754-2776 
               
               
                   
               
               
                 AD-1231104 
                 UUCAGUAUUUAUUUCUGUCUA 
                 198 
                 2757-2777 
                 UAGACAGAAAUAAAUACUGAAAG 
                 380 
                 2755-2777 
               
               
                   
               
               
                 AD-1231108 
                 AUUUGACUUCUGUUCUGUUUA 
                 199 
                 2781-2801 
                 UAAACAGAACAGAAGUCAAAUCC 
                 381 
                 2779-2801 
               
               
                   
               
               
                 AD-1231118 
                 UGCCUGAUAGAAACUCAUUUA 
                 200 
                 3236-3256 
                 UAAAUGAGUUUCUAUCAGGCAUG 
                 382 
                 3234-3256 
               
               
                   
               
            
           
         
       
     
     
       
         
           
               
             
               
                 TABLE 3 
               
             
            
               
                   
               
               
                 Modified Sense and Antisense Strand Human ACE2 dsRNA Sequences 
               
            
           
           
               
               
               
               
               
               
               
            
               
                   
                   
                 SEQ 
                   
                 SEQ 
                   
                 SEQ 
               
               
                 Duplex 
                 Sense 
                 ID 
                 Antisense 
                 ID 
                 mRNA Target 
                 ID 
               
               
                 Name 
                 Sequence 5′ to 3′ 
                 NO: 
                 Sequence 5′ to 3′ 
                 NO: 
                 Sequence 5′ to 3′ 
                 NO: 
               
               
                   
               
               
                 AD-1230821 
                 usasucaaAfgUfUf 
                 383 
                 VPusGfsaagCfaag 
                 560 
                 CUUAUCAAAGUUCAC 
                 737 
               
               
                   
                 Cfacu(Uhd)gcuus 
                   
                 ugaaCfuUfugauas 
                   
                 UUGCUUCU 
                   
               
               
                   
                 csa 
                   
                 gsa 
                   
                   
                   
               
               
                   
               
               
                 AD-1230822 
                 csusguu(Chd)Ufa 
                 384 
                 VPusAfsgugAfacu 
                 561 
                 ACCUGUCUUAUCAAA 
                 738 
               
               
                   
                 UfCfAfaaguucacs 
                   
                 uugaUfaGfaacags 
                   
                 GUUCACUU 
                   
               
               
                   
                 usa 
                   
                 gsu 
                   
                   
                   
               
               
                   
               
               
                 AD-1230823 
                 uscsuuc(Chd)Ufa 
                 385 
                 VPusCfsuugAfguc 
                 562 
                 GGCCUUCCUCAUAUG 
                 739 
               
               
                   
                 AfUfAfugacucaas 
                   
                 auauUfaGfgaagas 
                   
                 ACUCAAGG 
                   
               
               
                   
                 gsa 
                   
                 csc 
                   
                   
                   
               
               
                   
               
               
                 AD-1230825 
                 ususacu(Chd)Afu 
                 386 
                 VPusAfsauaUfcga 
                 563 
                 GAUUACUCAUUCAUU 
                 740 
               
               
                   
                 UfCfAfuucgauaus 
                   
                 augaAfuGfaguaas 
                   
                 CGAUAUUA 
                   
               
               
                   
                 usa 
                   
                 usc 
                   
                   
                   
               
               
                   
               
               
                 AD-1230826 
                 ascsuca(Uhd)Ufc 
                 387 
                 VPusGfsuaaUfauc 
                 564 
                 UUACUCAUUCAUUCG 
                 741 
               
               
                   
                 AfUfUfcgauauuas 
                   
                 gaauGfaAfugagus 
                   
                 AUAUUACA 
                   
               
               
                   
                 csa 
                   
                 asa 
                   
                   
                   
               
               
                   
               
               
                 AD-1230827 
                 asasug(Uhd)gAfc 
                 388 
                 VPusAfsgagUfuug 
                 565 
                 CAAAUGUGACAUCUC 
                 742 
               
               
                   
                 AfUfCfucaaacucs 
                   
                 agauGfuCfacauus 
                   
                 AAAUUCCA 
                   
               
               
                   
                 usa 
                   
                 usg 
                   
                   
                   
               
               
                   
               
               
                 AD-1230828 
                 asusguga(Chd)aU 
                 389 
                 VPusUfsagaGfuuu 
                 566 
                 AAAUGUGACAUCUCA 
                 743 
               
               
                   
                 fCfUfcaaacucusa 
                   
                 gagaUfgUfcacaus 
                   
                 AAUUCCAC 
                   
               
               
                   
                 sa 
                   
                 usu 
                   
                   
                   
               
               
                   
               
               
                 AD-1230829 
                 usgsaaa(Chd)Cfa 
                 390 
                 VPusAfsaagGfaga 
                 567 
                 UUUGAAACCAAGAGU 
                 744 
               
               
                   
                 AfGfAfaucuccuus 
                   
                 uucuUfgGfuuucas 
                   
                 CUCCUUCU 
                   
               
               
                   
                 usa 
                   
                 asa 
                   
                   
                   
               
               
                   
               
               
                 AD-1230831 
                 ususgga(Chd)Afa 
                 391 
                 VPusAfsagaGfuac 
                 568 
                 UUUUGGACAAAUCUG 
                 745 
               
               
                   
                 AfUfCfuguacucus 
                   
                 agauUfuGfuccaas 
                   
                 UACCCUUU 
                   
               
               
                   
                 usa 
                   
                 asa 
                   
                   
                   
               
               
                   
               
               
                 AD-1230832 
                 asasauc(Uhd)Gfu 
                 392 
                 VPusCfsuguCfaaa 
                 569 
                 ACAAAUCUGUACCCU 
                 746 
               
               
                   
                 AfCfUfcuuugacas 
                   
                 gaguAfcAfgauuus 
                   
                 UUGACUGU 
                   
               
               
                   
                 gsa 
                   
                 gsu 
                   
                   
                   
               
               
                   
               
               
                 AD-1230833 
                 ususcuu(Uhd)Gfu 
                 393 
                 VPusAfsgacCfaac 
                 570 
                 AAUUCUUUGUUUCUG 
                 747 
               
               
                   
                 AfUfCfuguuggucs 
                   
                 agauAfcAfaagaas 
                   
                 UUGGCCUU 
                   
               
               
                   
                 usa 
                   
                 csu 
                   
                   
                   
               
               
                   
               
               
                 AD-1230836 
                 uscsucug(Uhd)uC 
                 394 
                 VPusUfsuagAfaac 
                 571 
                 CAUCUCUGUUCCAUG 
                 748 
               
               
                   
                 fCfAfuguuucuasa 
                   
                 auggAfaCfagagas 
                   
                 UUUCUAAU 
                   
               
               
                   
                 sa 
                   
                 usg 
                   
                   
                   
               
               
                   
               
               
                 AD-1230837 
                 uscsugu(Uhd)Cfc 
                 395 
                 VPusCfsauuAfgaa 
                 572 
                 UCUCUGUUCCAUGUU 
                 749 
               
               
                   
                 AfUfGfuuucuaaus 
                   
                 acauGfgAfacagas 
                   
                 UCUAAUGA 
                   
               
               
                   
                 gsa 
                   
                 gsa 
                   
                   
                   
               
               
                   
               
               
                 AD-1230838 
                 ususcca(Uhd)Gfu 
                 396 
                 VPusUfsaauCfauu 
                 573 
                 UGUUCCAUGUUUCUA 
                 750 
               
               
                   
                 UfUfCfuaaugauus 
                   
                 agaaAfcAfuggaas 
                   
                 AUGAUUAC 
                   
               
               
                   
                 asa 
                   
                 csa 
                   
                   
                   
               
               
                   
               
               
                 AD-1230839 
                 uscscaug(Uhd)uU 
                 397 
                 VPusGfsuaaUfcau 
                 574 
                 GUUCCAUGUUUCUAA 
                 751 
               
               
                   
                 fCfUfaaugauuasc 
                   
                 uagaAfaCfauggas 
                   
                 UGAUUACU 
                   
               
               
                   
                 sa 
                   
                 asc 
                   
                   
                   
               
               
                   
               
               
                 AD-1230841 
                 usasauga(Uhd)uA 
                 398 
                 VPusGfsaauGfaau 
                 575 
                 UCUAAUGAUUACUCA 
                 752 
               
               
                   
                 fCfUfcauucauusc 
                   
                 gaguAfaUfcauuas 
                   
                 UUCAUUCG 
                   
               
               
                   
                 sa 
                   
                 gsa 
                   
                   
                   
               
               
                   
               
               
                 AD-1230842 
                 gsasuua(Chd)Ufc 
                 399 
                 VPusUfsaucGfaau 
                 576 
                 AUGAUUACUCAUUCA 
                 753 
               
               
                   
                 AfUfUfcauucgaus 
                   
                 gaauGfaGfuaaucs 
                   
                 UUCGAUAU 
                   
               
               
                   
                 asa 
                   
                 asu 
                   
                   
                   
               
               
                   
               
               
                 AD-1230843 
                 csuscau(Uhd)Cfa 
                 400 
                 VPusUfsguaAfuau 
                 577 
                 UACUCAUUCAUUCGA 
                 754 
               
               
                   
                 UfUfCfgauauuacs 
                   
                 cgaaUfgAfaugags 
                   
                 UAUUACAC 
                   
               
               
                   
                 asa 
                   
                 usa 
                   
                   
                   
               
               
                   
               
               
                 AD-1230844 
                 gsasccug(Uhd)uC 
                 401 
                 VPusGfsaacUfuug 
                 578 
                 AAGACCUGUCUUAUC 
                 755 
               
               
                   
                 fUfAfucaaaguusc 
                   
                 auagAfaCfaggucs 
                   
                 AAAGUUCA 
                   
               
               
                   
                 sa 
                   
                 usu 
                   
                   
                   
               
               
                   
               
               
                 AD-1230845 
                 cscsuuua(Chd)cA 
                 402 
                 VPusGfsaaaCfugg 
                 579 
                 ACCAUUUACCAAUUC 
                 756 
               
               
                   
                 fAfUfuccaguuusc 
                   
                 aauuGfgUfaaaggs 
                   
                 CAGUUUCA 
                   
               
               
                   
                 sa 
                   
                 gsu 
                   
                   
                   
               
               
                   
               
               
                 AD-1230846 
                 escsugu(Uhd)Cfu 
                 403 
                 VPusGfsugaAfcuu 
                 580 
                 GACCUGUCUUAUCAA 
                 757 
               
               
                   
                 AfUfCfaaaguucas 
                   
                 ugauAfgAfacaggs 
                   
                 AGUUCACU 
                   
               
               
                   
                 csa 
                   
                 usc 
                   
                   
                   
               
               
                   
               
               
                 AD-1230847 
                 usgsuuc(Uhd)Afu 
                 404 
                 VPusAfsaguGfaac 
                 581 
                 CCUGUCUUAUCAAAG 
                 758 
               
               
                   
                 CfAfAfaguucacus 
                   
                 uuugAfuAfgaacas 
                   
                 UUCACUUG 
                   
               
               
                   
                 usa 
                   
                 gsg 
                   
                   
                   
               
               
                   
               
               
                 AD-1230848 
                 gsusucua(Uhd)cA 
                 405 
                 VPusCfsaagUfgaa 
                 582 
                 CUGUCUUAUCAAAGU 
                 759 
               
               
                   
                 fAfAfguucacuusg 
                   
                 cuuuGfaUfagaacs 
                   
                 UCACUUGC 
                   
               
               
                   
                 sa 
                   
                 asg 
                   
                   
                   
               
               
                   
               
               
                 AD-1230849 
                 uscsuau(Chd)Afa 
                 406 
                 VPusAfsgcaAfgug 
                 583 
                 GUCUUAUCAAAGUUC 
                 760 
               
               
                   
                 AfGfUfucacuugcs 
                   
                 aacuUfuGfauagas 
                   
                 ACUUGCUU 
                   
               
               
                   
                 usa 
                   
                 asc 
                   
                   
                   
               
               
                   
               
               
                 AD-1230850 
                 asasaug(Uhd)Gfa 
                 407 
                 VPusGfsaguUfuga 
                 584 
                 ACAAAUGUGACAUCU 
                 761 
               
               
                   
                 CfAfUfcucaaacus 
                   
                 gaugUfcAfcauuus 
                   
                 CAAAUUCC 
                   
               
               
                   
                 csa 
                   
                 gsu 
                   
                   
                   
               
               
                   
               
               
                 AD-1230851 
                 csusau(Chd)aAfa 
                 408 
                 VPusAfsagcAfagu 
                 585 
                 UCUUAUCAAAGUUCA 
                 762 
               
               
                   
                 GfUfUfcacuugcus 
                   
                 gaacUfuUfgauags 
                   
                 CUUGCUUC 
                   
               
               
                   
                 usa 
                   
                 asa 
                   
                   
                   
               
               
                   
               
               
                 AD-1230852 
                 usgsuga(Chd)Afu 
                 409 
                 VPusGfsuagAfguu 
                 586 
                 AAUGUGACAUCUCAA 
                 763 
               
               
                   
                 CfUfCfaaacucuas 
                   
                 ugagAfuGfucacas 
                   
                 AUUCCACU 
                   
               
               
                   
                 csa 
                   
                 usu 
                   
                   
                   
               
               
                   
               
               
                 AD-1230853 
                 csasucu(Chd)Afa 
                 410 
                 VPusCfsuucUfgua 
                 587 
                 GACAUCUCAAAUUCC 
                 764 
               
               
                   
                 AfCfUfcuacagaas 
                   
                 gaguUfuGfagaugs 
                   
                 ACUGAAGC 
                   
               
               
                   
                 gsa 
                   
                 usc 
                   
                   
                   
               
               
                   
               
               
                 AD-1230855 
                 asuscaaaGfuUfCf 
                 411 
                 VPusAfsgaaGfcaa 
                 588 
                 UUAUCAAAGUUCACU 
                 765 
               
               
                   
                 Afcuug(Chd)uucs 
                   
                 gugaAfcUfuugaus 
                   
                 UGCUUCUU 
                   
               
               
                   
                 usa 
                   
                 asg 
                   
                   
                   
               
               
                   
               
               
                 AD-1230856 
                 uscsaaag(Uhd)uC 
                 412 
                 VPusAfsagaAfgca 
                 589 
                 UAUCAAAGUUCACUU 
                 766 
               
               
                   
                 fAfCfuugcuucusu 
                   
                 agugAfaCfuuugas 
                   
                 GCUUCUUG 
                   
               
               
                   
                 sa 
                   
                 usa 
                   
                   
                   
               
               
                   
               
               
                 AD-1230857 
                 asasccaaGfaAfUf 
                 413 
                 VPusAfsuuaAfagg 
                 590 
                 GAAACCAAGAGUCUC 
                 767 
               
               
                   
                 Cfuccu(Uhd)uaas 
                   
                 agauUfcUfugguus 
                   
                 CUUCUACU 
                   
               
               
                   
                 usa 
                   
                 usc 
                   
                   
                   
               
               
                   
               
               
                 AD-1230858 
                 ascscaagAfaUfCf 
                 414 
                 VPusAfsauuAfaag 
                 591 
                 AAACCAAGAGUCUCC 
                 768 
               
               
                   
                 Ufccuu(Uhd)aaus 
                   
                 gagaUfuCfuuggus 
                   
                 UUCUACUU 
                   
               
               
                   
                 usa 
                   
                 usu 
                   
                   
                   
               
               
                   
               
               
                 AD-1230859 
                 uscsauu(Chd)Cfu 
                 415 
                 VPusAfsacuUfcag 
                 592 
                 UGUCAUUCCUAGAAG 
                 769 
               
               
                   
                 AfGfAfacugaagus 
                   
                 uucuAfgGfaaugas 
                   
                 UGAAGUUG 
                   
               
               
                   
                 usa 
                   
                 usa 
                   
                   
                   
               
               
                   
               
               
                 AD-1230867 
                 csgsgu(Uhd)gAfa 
                 416 
                 VPusUfsuuaGfaau 
                 593 
                 AACAGUUGAACACAA 
                 770 
               
               
                   
                 CfAfCfaauucuaas 
                   
                 ugugUfuCfaaccgs 
                   
                 UUCUGAAC 
                   
               
               
                   
                 asa 
                   
                 usu 
                   
                   
                   
               
               
                   
               
               
                 AD-1230868 
                 gsgsuugaAfcAfCf 
                 417 
                 VPusAfsuuuAfgaa 
                 594 
                 ACAGUUGAACACAAU 
                 771 
               
               
                   
                 Afauuc(Uhd)aaas 
                   
                 uuguGfuUfcaaccs 
                   
                 UCUGAACA 
                   
               
               
                   
                 usa 
                   
                 gsu 
                   
                   
                   
               
               
                   
               
               
                 AD-1230869 
                 gsusugaa(Chd)aC 
                 418 
                 VPusUfsauuUfaga 
                 595 
                 CAGUUGAACACAAUU 
                 772 
               
               
                   
                 fAfAfuucuaaausa 
                   
                 auugUfgUfucaacs 
                   
                 CUGAACAC 
                   
               
               
                   
                 sa 
                   
                 csg 
                   
                   
                   
               
               
                   
               
               
                 AD-1230870 
                 usgsga(Chd)aAfa 
                 419 
                 VPusAfsaagAfgua 
                 596 
                 UUUGGACAAAUCUGU 
                 773 
               
               
                   
                 UfCfUfguacucuus 
                   
                 cagaUfuUfguccas 
                   
                 ACCCUUUG 
                   
               
               
                   
                 usa 
                   
                 asa 
                   
                   
                   
               
               
                   
               
               
                 AD-1230871 
                 csusucc(Uhd)Afa 
                 420 
                 VPusCfscuuGfagu 
                 597 
                 GCCUUCCUCAUAUGA 
                 774 
               
               
                   
                 UfAfUfgacucaags 
                   
                 cauaUfuAfggaags 
                   
                 CUCAAGGA 
                   
               
               
                   
                 gsa 
                   
                 asc 
                   
                   
                   
               
               
                   
               
               
                 AD-1230874 
                 asasccu(Uhd)Ufu 
                 421 
                 VPusAfsuuuCfuua 
                 598 
                 GCAACCUUUCCUGCU 
                 775 
               
               
                   
                 CfUfGfcuaagaaas 
                   
                 gcagAfaAfagguus 
                   
                 AAGAAACG 
                   
               
               
                   
                 usa 
                   
                 gsu 
                   
                   
                   
               
               
                   
               
               
                 AD-1230877 
                 csusguu(Chd)Cfa 
                 422 
                 VPusUfscauUfaga 
                 599 
                 CUCUGUUCCAUGUUU 
                 776 
               
               
                   
                 UfGfUfuucuaaugs 
                   
                 aacaUfgGfaacags 
                   
                 CUAAUGAU 
                   
               
               
                   
                 asa 
                   
                 asg 
                   
                   
                   
               
               
                   
               
               
                 AD-1230878 
                 usgsuuc(Chd)Afu 
                 423 
                 VPusAfsucaUfuag 
                 600 
                 UCUGUUCCAUGUUUC 
                 777 
               
               
                   
                 GfUfUfucuaaugas 
                   
                 aaacAfuGfgaacas 
                   
                 UAAUGAUU 
                   
               
               
                   
                 usa 
                   
                 gsa 
                   
                   
                   
               
               
                   
               
               
                 AD-1230879 
                 gsusucca(Uhd)gU 
                 424 
                 VPusAfsaucAfuua 
                 601 
                 CUGUUCCAUGUUUCU 
                 778 
               
               
                   
                 fUfUfcuaaugausu 
                   
                 gaaaCfaUfggaacs 
                   
                 AAUGAUUA 
                   
               
               
                   
                 sa 
                   
                 asg 
                   
                   
                   
               
               
                   
               
               
                 AD-1230880 
                 ususcuaa(Uhd)gA 
                 425 
                 VPusUfsgaaUfgag 
                 602 
                 GUUUCUAAUGAUUAC 
                 779 
               
               
                   
                 fUfUfacucauucsa 
                   
                 uaauCfaUfuagaas 
                   
                 UCAUUCAU 
                   
               
               
                   
                 sa 
                   
                 asc 
                   
                   
                   
               
               
                   
               
               
                 AD-1230881 
                 asasuga(Uhd)Ufa 
                 426 
                 VPusCfsgaaUfgaa 
                 603 
                 CUAAUGAUUACUCAU 
                 780 
               
               
                   
                 CfUfCfauucauucs 
                   
                 ugagUfaAfucauus 
                   
                 UCAUUCGA 
                   
               
               
                   
                 gsa 
                   
                 asg 
                   
                   
                   
               
               
                   
               
               
                 AD-1230882 
                 asusuac(Uhd)Cfa 
                 427 
                 VPusAfsuauCfgaa 
                 604 
                 UGAUUACUCAUUCAU 
                 781 
               
               
                   
                 UfUfCfauucgauas 
                   
                 ugaaUfgAfguaaus 
                   
                 UCGAUAUU 
                   
               
               
                   
                 usa 
                   
                 csa 
                   
                   
                   
               
               
                   
               
               
                 AD-1230883 
                 uscsauu(Chd)Afu 
                 428 
                 VPusGfsuguAfaua 
                 605 
                 ACUCAUUCAUUCGAU 
                 782 
               
               
                   
                 UfCfGfauauuacas 
                   
                 ucgaAfuGfaaugas 
                   
                 AUUACACA 
                   
               
               
                   
                 csa 
                   
                 gsu 
                   
                   
                   
               
               
                   
               
               
                 AD-1230884 
                 csasuuca(Uhd)uC 
                 429 
                 VPusUfsgugUfaau 
                 606 
                 CUCAUUCAUUCGAUA 
                 783 
               
               
                   
                 fGfAfuauuacacsa 
                   
                 aucgAfaUfgaaugs 
                   
                 UUACACAA 
                   
               
               
                   
                 sa 
                   
                 asg 
                   
                   
                   
               
               
                   
               
               
                 AD-1230885 
                 asgsacc(Uhd)Gfu 
                 430 
                 VPusAfsacuUfuga 
                 607 
                 GAAGACCUGUCUUAU 
                 784 
               
               
                   
                 UfCfUfaucaaagus 
                   
                 uagaAfcAfggucus 
                   
                 CAAAGUUC 
                   
               
               
                   
                 usa 
                   
                 usc 
                   
                   
                   
               
               
                   
               
               
                 AD-1230886 
                 ascscug(Uhd)Ufc 
                 431 
                 VPusUfsgaaCfuuu 
                 608 
                 AGACCUGUCUUAUCA 
                 785 
               
               
                   
                 UfAfUfcaaaguucs 
                   
                 gauaGfaAfcaggus 
                   
                 AAGUUCAC 
                   
               
               
                   
                 asa 
                   
                 csu 
                   
                   
                   
               
               
                   
               
               
                 AD-1230887 
                 csusuua(Chd)Cfa 
                 432 
                 VPusUfsgaaAfcug 
                 609 
                 CCAUUUACCAAUUCC 
                 786 
               
               
                   
                 AfUfUfccaguuucs 
                   
                 gaauUfgGfuaaags 
                   
                 AGUUUCAA 
                   
               
               
                   
                 asa 
                   
                 gsg 
                   
                   
                   
               
               
                   
               
               
                 AD-1230888 
                 ususac(Chd)aAfu 
                 433 
                 VPusCfsuugAfaac 
                 610 
                 AUUUACCAAUUCCAG 
                 787 
               
               
                   
                 UfCfCfaguuucaas 
                   
                 uggaAfuUfgguaas 
                   
                 UUUCAAGA 
                   
               
               
                   
                 gsa 
                   
                 asg 
                   
                   
                   
               
               
                   
               
               
                 AD-1230889 
                 ususcaagAfaGfCf 
                 434 
                 VPusUfsugaCfaaa 
                 611 
                 GUUUCAAGAAGCUCU 
                 788 
               
               
                   
                 Afcuu(Uhd)gucas 
                   
                 gugcUfuCfuugaas 
                   
                 UUGUCAAG 
                   
               
               
                   
                 asa 
                   
                 asc 
                   
                   
                   
               
               
                   
               
               
                 AD-1230890 
                 csasaaug(Uhd)gA 
                 435 
                 VPusAfsguuUfgag 
                 612 
                 CACAAAUGUGACAUC 
                 789 
               
               
                   
                 fCfAfucucaaacsu 
                   
                 auguCfaCfauuugs 
                   
                 UCAAAUUC 
                   
               
               
                   
                 sa 
                   
                 usg 
                   
                   
                   
               
               
                   
               
               
                 AD-1230891 
                 usgsaca(Uhd)Cfu 
                 436 
                 VPusCfsuguAfgag 
                 613 
                 UGUGACAUCUCAAAU 
                 790 
               
               
                   
                 CfAfAfacucuacas 
                   
                 uuugAfgAfugucas 
                   
                 UCCACUGA 
                   
               
               
                   
                 gsa 
                   
                 csa 
                   
                   
                   
               
               
                   
               
               
                 AD-1230892 
                 gsascau(Chd)Ufc 
                 437 
                 VPusUfscugUfaga 
                 614 
                 GUGACAUCUCAAAUU 
                 791 
               
               
                   
                 AfAfAfcucuacags 
                   
                 guuuGfaGfaugucs 
                   
                 CCACUGAA 
                   
               
               
                   
                 asa 
                   
                 asc 
                   
                   
                   
               
               
                   
               
               
                 AD-1230894 
                 csasaag(Uhd)Ufc 
                 438 
                 VPusCfsaagAfagc 
                 615 
                 AUCAAAGUUCACUUG 
                 792 
               
               
                   
                 AfCfUfugcuucuus 
                   
                 aaguGfaAfcuuugs 
                   
                 CUUCUUGG 
                   
               
               
                   
                 gsa 
                   
                 asu 
                   
                   
                   
               
               
                   
               
               
                 AD-1230895 
                 asasagu(Uhd)Cfa 
                 439 
                 VPusCfscaaGfaag 
                 616 
                 UCAAAGUUCACUUGC 
                 793 
               
               
                   
                 CfUfUfgcuucuugs 
                   
                 caagUfgAfacuuus 
                   
                 UUCUUGGA 
                   
               
               
                   
                 gsa 
                   
                 gsa 
                   
                   
                   
               
               
                   
               
               
                 AD-1230896 
                 gsusuca(Chd)Ufu 
                 440 
                 VPusAfsuucCfaag 
                 617 
                 AAGUUCACUUGCUUC 
                 794 
               
               
                   
                 GfCfUfucuuggaas 
                   
                 aagcAfaGfugaacs 
                   
                 UUGGAAUU 
                   
               
               
                   
                 usa 
                   
                 usu 
                   
                   
                   
               
               
                   
               
               
                 AD-1230897 
                 ususgcu(Uhd)Cfu 
                 441 
                 VPusGfsuuaUfaau 
                 618 
                 ACUUGCUUCUUGGAA 
                 795 
               
               
                   
                 UfGfGfaauuauaas 
                   
                 uccaAfgAfagcaas 
                   
                 UUAUAAUA 
                   
               
               
                   
                 csa 
                   
                 gsu 
                   
                   
                   
               
               
                   
               
               
                 AD-1230898 
                 cscsaagaAfuCfUf 
                 442 
                 VPusAfsaauUfaaa 
                 619 
                 AACCAAGAGUCUCCU 
                 796 
               
               
                   
                 Cfcuu(Uhd)aauus 
                   
                 ggagAfuUfcuuggs 
                   
                 UCUACUUC 
                   
               
               
                   
                 usa 
                   
                 usu 
                   
                   
                   
               
               
                   
               
               
                 AD-1230902 
                 ususuga(Chd)Ufu 
                 443 
                 VPusGfsaaaCfaga 
                 620 
                 GAUUUGACAUCUCUU 
                 797 
               
               
                   
                 CfUfGfuucuguuus 
                   
                 acagAfaGfucaaas 
                   
                 CUGUUUAU 
                   
               
               
                   
                 csa 
                   
                 usc 
                   
                   
                   
               
               
                   
               
               
                 AD-1230904 
                 csasagaaAfuUfCf 
                 444 
                 VPusGfsugaGfauu 
                 621 
                 UACAAGAAAUCCAGA 
                 798 
               
               
                   
                 Afgaau(Chd)ucas 
                   
                 cugaAfuUfucuugs 
                   
                 CUCCGAUC 
                   
               
               
                   
                 csa 
                   
                 usa 
                   
                   
                   
               
               
                   
               
               
                 AD-1230910 
                 gsgsacaaAfuCfUf 
                 445 
                 VPusCfsaaaGfagu 
                 622 
                 UUGGACAAAUCUGUA 
                 799 
               
               
                   
                 Gfuacu(Chd)uuus 
                   
                 acagAfuUfuguccs 
                   
                 CCCUUUGA 
                   
               
               
                   
                 gsa 
                   
                 asa 
                   
                   
                   
               
               
                   
               
               
                 AD-1230911 
                 asasucug(Uhd)aC 
                 446 
                 VPusAfscugUfcaa 
                 623 
                 CAAAUCUGUACCCUU 
                 800 
               
               
                   
                 fUfCfuuugacagsu 
                   
                 agagUfaCfagauus 
                   
                 UGACUGUU 
                   
               
               
                   
                 sa 
                   
                 usg 
                   
                   
                   
               
               
                   
               
               
                 AD-1230912 
                 uscsugua(Chd)uC 
                 447 
                 VPusGfsaacUfguc 
                 624 
                 AAUCUGUACCCUUUG 
                 801 
               
               
                   
                 fUfUfugacaguusc 
                   
                 aaagAfgUfacagas 
                   
                 ACUGUUCC 
                   
               
               
                   
                 sa 
                   
                 usu 
                   
                   
                   
               
               
                   
               
               
                 AD-1230913 
                 uscsuuug(Uhd)aU 
                 448 
                 VPusAfsagaCfcaa 
                 625 
                 AUUCUUUGUUUCUGU 
                 802 
               
               
                   
                 fCfUfguuggucusu 
                   
                 cagaUfaCfaaagas 
                   
                 UGGCCUUC 
                   
               
               
                   
                 sa 
                   
                 asc 
                   
                   
                   
               
               
                   
               
               
                 AD-1230914 
                 ususcc(Uhd)aAfu 
                 449 
                 VPusUfsccuUfgag 
                 626 
                 CCUUCCUCAUAUGAC 
                 803 
               
               
                   
                 AfUfGfacucaaggs 
                   
                 ucauAfuUfaggaas 
                   
                 UCAAGGAU 
                   
               
               
                   
                 asa 
                   
                 gsa 
                   
                   
                   
               
               
                   
               
               
                 AD-1230915 
                 uscscuaa(Uhd)aU 
                 450 
                 VPusAfsuccUfuga 
                 627 
                 CUUCCUCAUAUGACU 
                 804 
               
               
                   
                 fGfAfcucaaggasu 
                   
                 gucaUfaUfuaggas 
                   
                 CAAGGAUU 
                   
               
               
                   
                 sa 
                   
                 asg 
                   
                   
                   
               
               
                   
               
               
                 AD-1230916 
                 asasgga(Uhd)Ufc 
                 451 
                 VPusGfsgaaUfuuu 
                 628 
                 UCAAGGAUUCUGGGC 
                 805 
               
               
                   
                 UfGfGfgaaaauucs 
                   
                 cccaGfaAfuccuus 
                   
                 AAACUCUA 
                   
               
               
                   
                 csa 
                   
                 gsa 
                   
                   
                   
               
               
                   
               
               
                 AD-1230920 
                 csasuu(Uhd)aAfa 
                 452 
                 VPusAfsgacCfaau 
                 629 
                 AGCAUCUGAAAUCCA 
                 806 
               
               
                   
                 AfUfCfcauuggucs 
                   
                 ggauUfuUfaaaugs 
                   
                 UUGGUCUU 
                   
               
               
                   
                 usa 
                   
                 csu 
                   
                   
                   
               
               
                   
               
               
                 AD-1230921 
                 asasaau(Chd)Cfa 
                 453 
                 VPusAfscagAfaga 
                 630 
                 CUGAAAUCCAUUGGU 
                 807 
               
               
                   
                 UfUfGfgucuucugs 
                   
                 ccaaUfgGfauuuus 
                   
                 CUUCUGCC 
                   
               
               
                   
                 usa 
                   
                 asa 
                   
                   
                   
               
               
                   
               
               
                 AD-1230922 
                 asasauc(Chd)Afu 
                 454 
                 VPusGfsacaGfaag 
                 631 
                 UGAAAUCCAUUGGUC 
                 808 
               
               
                   
                 UfGfGfucuucugus 
                   
                 accaAfuGfgauuus 
                   
                 UUCUGCCA 
                   
               
               
                   
                 csa 
                   
                 usa 
                   
                   
                   
               
               
                   
               
               
                 AD-1230923 
                 asasucca(Uhd)uG 
                 455 
                 VPusUfsgacAfgaa 
                 632 
                 GAAAUCCAUUGGUCU 
                 809 
               
               
                   
                 fGfUfcuucugucsa 
                   
                 gaccAfaUfggauus 
                   
                 UCUGCCAU 
                   
               
               
                   
                 sa 
                   
                 usu 
                   
                   
                   
               
               
                   
               
               
                 AD-1230928 
                 gscscau(Uhd)Ufa 
                 456 
                 VPusCfsuaaCfaug 
                 633 
                 CUACCGUUUACUUAC 
                 810 
               
               
                   
                 CfUfUfacauguuas 
                   
                 uaagUfaAfauggcs 
                   
                 AUGUUAGA 
                   
               
               
                   
                 gsa 
                   
                 asg 
                   
                   
                   
               
               
                   
               
               
                 AD-1230930 
                 asuscuc(Uhd)Gfu 
                 457 
                 VPusUfsagaAfaca 
                 634 
                 GCAUCUCUGUUCCAU 
                 811 
               
               
                   
                 UfCfCfauguuucus 
                   
                 uggaAfcAfgagaus 
                   
                 GUUUCUAA 
                   
               
               
                   
                 asa 
                   
                 gsc 
                   
                   
                   
               
               
                   
               
               
                 AD-1230931 
                 csuscug(Uhd)Ufc 
                 458 
                 VPusAfsuuaGfaaa 
                 635 
                 AUCUCUGUUCCAUGU 
                 812 
               
               
                   
                 CfAfUfguuucuaas 
                   
                 caugGfaAfcagags 
                   
                 UUCUAAUG 
                   
               
               
                   
                 usa 
                   
                 asu 
                   
                   
                   
               
               
                   
               
               
                 AD-1230932 
                 escsaug(Uhd)Ufu 
                 459 
                 VPusAfsguaAfuca 
                 636 
                 UUCCAUGUUUCUAAU 
                 813 
               
               
                   
                 CfUfAfaugauuacs 
                   
                 uuagAfaAfcauggs 
                   
                 GAUUACUC 
                   
               
               
                   
                 usa 
                   
                 asa 
                   
                   
                   
               
               
                   
               
               
                 AD-1230934 
                 usgsauua(Chd)uC 
                 460 
                 VPusAfsucgAfaug 
                 637 
                 AAUGAUUACUCAUUC 
                 814 
               
               
                   
                 fAfUfucauucgasu 
                   
                 aaugAfgUfaaucas 
                   
                 AUUCGAUA 
                   
               
               
                   
                 sa 
                   
                 usu 
                   
                   
                   
               
               
                   
               
               
                 AD-1230935 
                 ususuac(Chd)Afa 
                 461 
                 VPusUfsugaAfacu 
                 638 
                 CAUUUACCAAUUCCA 
                 815 
               
               
                   
                 UfUfCfcaguuucas 
                   
                 ggaaUfuGfguaaas 
                   
                 GUUUCAAG 
                   
               
               
                   
                 asa 
                   
                 gsg 
                   
                   
                   
               
               
                   
               
               
                 AD-1230936 
                 gscsacaaAfuGfUf 
                 462 
                 VPusUfsugaGfaug 
                 639 
                 CUGCACAAAUGUGAC 
                 816 
               
               
                   
                 Gfacau(Chd)ucas 
                   
                 ucacAfuUfugugcs 
                   
                 AUCUCAAA 
                   
               
               
                   
                 asa 
                   
                 asg 
                   
                   
                   
               
               
                   
               
               
                 AD-1230937 
                 asgsuuca(Chd)uU 
                 463 
                 VPusUfsuccAfaga 
                 640 
                 AAAGUUCACUUGCUU 
                 817 
               
               
                   
                 fGfCfuucuuggasa 
                   
                 agcaAfgUfgaacus 
                   
                 CUUGGAAU 
                   
               
               
                   
                 sa 
                   
                 usu 
                   
                   
                   
               
               
                   
               
               
                 AD-1230938 
                 usgscuu(Chd)Ufu 
                 464 
                 VPusUfsguuAfuaa 
                 641 
                 CUUGCUUCUUGGAAU 
                 818 
               
               
                   
                 GfGfAfauuauaacs 
                   
                 uuccAfaGfaagcas 
                   
                 UAUAAUAC 
                   
               
               
                   
                 asa 
                   
                 asg 
                   
                   
                   
               
               
                   
               
               
                 AD-1230939 
                 usgsgaa(Uhd)Ufa 
                 465 
                 VPusAfsuauUfggu 
                 642 
                 CUUGGAAUUAUAAUA 
                 819 
               
               
                   
                 UfAfAfcaccaauas 
                   
                 guuaUfaAfuuccas 
                   
                 CUAACAUU 
                   
               
               
                   
                 usa 
                   
                 asg 
                   
                   
                   
               
               
                   
               
               
                 AD-1230940 
                 ususgaaa(Chd)cA 
                 466 
                 VPusAfsaggAfgau 
                 643 
                 AUUUGAAACCAAGAG 
                 820 
               
               
                   
                 fAfGfaaucuccusu 
                   
                 ucuuGfgUfuucaas 
                   
                 UCUCCUUC 
                   
               
               
                   
                 sa 
                   
                 asu 
                   
                   
                   
               
               
                   
               
               
                 AD-1230941 
                 asasac(Chd)aAfg 
                 467 
                 VPusUfsuaaAfgga 
                 644 
                 UGAAACCAAGAGUCU 
                 821 
               
               
                   
                 AfAfUfcuccuuuas 
                   
                 gauuCfuUfgguuus 
                   
                 CCUUCUAC 
                   
               
               
                   
                 asa 
                   
                 csa 
                   
                   
                   
               
               
                   
               
               
                 AD-1230942 
                 usasuca(Uhd)Ufc 
                 468 
                 VPusCfsuucAfguu 
                 645 
                 GAUGUCAUUCCUAGA 
                 822 
               
               
                   
                 CfUfAfgaacugaas 
                   
                 cuagGfaAfugauas 
                   
                 AGUGAAGU 
                   
               
               
                   
                 gsa 
                   
                 usc 
                   
                   
                   
               
               
                   
               
               
                 AD-1230946 
                 asusccagGfaUfUf 
                 469 
                 VPusAfsgugUfuuu 
                 646 
                 CAAUGCAGGAUUCCA 
                 823 
               
               
                   
                 Cfcaaaa(Chd)acs 
                   
                 ggaaUfcCfuggaus 
                   
                 AAACAGUG 
                   
               
               
                   
                 usa 
                   
                 usa 
                   
                   
                   
               
               
                   
               
               
                 AD-1230947 
                 ascscuc(Chd)Ufu 
                 470 
                 VPusAfsgauUfuuu 
                 647 
                 AGACUUCCUUUUAGC 
                 824 
               
               
                   
                 UfUfAfgaaaaaucs 
                   
                 cuaaAfaGfgaggus 
                   
                 AAAGCACU 
                   
               
               
                   
                 usa 
                   
                 csu 
                   
                   
                   
               
               
                   
               
               
                 AD-1230948 
                 cscsucc(Uhd)Ufu 
                 471 
                 VPusUfsagaUfuuu 
                 648 
                 GACUUCCUUUUAGCA 
                 825 
               
               
                   
                 UfAfGfaaaaaucus 
                   
                 ucuaAfaAfggaggs 
                   
                 AAGCACUU 
                   
               
               
                   
                 asa 
                   
                 usc 
                   
                   
                   
               
               
                   
               
               
                 AD-1230949 
                 asuscuu(Chd)Afu 
                 472 
                 VPusAfsaagCfaau 
                 649 
                 ACAUCUUCACUGACA 
                 826 
               
               
                   
                 UfGfAfcauugcuus 
                   
                 gucaAfuGfaagaus 
                   
                 UUGCUUUC 
                   
               
               
                   
                 usa 
                   
                 gsc 
                   
                   
                   
               
               
                   
               
               
                 AD-1230950 
                 uscsuuca(Uhd)uG 
                 473 
                 VPusGfsaaaGfcaa 
                 650 
                 CAUCUUCACUGACAU 
                 827 
               
               
                   
                 fAfCfauugcuuusc 
                   
                 ugucAfaUfgaagas 
                   
                 UGCUUUCA 
                   
               
               
                   
                 sa 
                   
                 usg 
                   
                   
                   
               
               
                   
               
               
                 AD-1230951 
                 asgsuau(Uhd)Ufa 
                 474 
                 VPusCfsagaGfaca 
                 651 
                 UCAGUAUUUAUUUCU 
                 828 
               
               
                   
                 UfUfUfcugucucus 
                   
                 gaaaUfaAfauacus 
                   
                 GCCUAAGG 
                   
               
               
                   
                 gsa 
                   
                 gsa 
                   
                   
                   
               
               
                   
               
               
                 AD-1230962 
                 ascsaagaAfaUfUf 
                 475 
                 VPusUfsgagAfuuc 
                 652 
                 CUACAAGAAAUCCAG 
                 829 
               
               
                   
                 Cfagaa(Uhd)cucs 
                   
                 ugaaUfuUfcuugus 
                   
                 ACUCCGAU 
                   
               
               
                   
                 asa 
                   
                 asg 
                   
                   
                   
               
               
                   
               
               
                 AD-1230969 
                 ascsggu(Uhd)Gfa 
                 476 
                 VPusUfsuagAfauu 
                 653 
                 AAACAGUUGAACACA 
                 830 
               
               
                   
                 AfCfAfcaauucuas 
                   
                 guguUfcAfaccgus 
                   
                 AUUCUGAA 
                   
               
               
                   
                 asa 
                   
                 usu 
                   
                   
                   
               
               
                   
               
               
                 AD-1230970 
                 ususgaa(Chd)Afc 
                 477 
                 VPusGfsuauUfuag 
                 654 
                 AGUUGAACACAAUUC 
                 831 
               
               
                   
                 AfAfUfucuaaauas 
                   
                 aauuGfuGfuucaas 
                   
                 UGAACACC 
                   
               
               
                   
                 csa 
                   
                 csc 
                   
                   
                   
               
               
                   
               
               
                 AD-1230971 
                 usgsaaca(Chd)aA 
                 478 
                 VPusUfsguaUfuua 
                 655 
                 GUUGAACACAAUUCU 
                 832 
               
               
                   
                 fUfUfcuaaauacsa 
                   
                 gaauUfgUfguucas 
                   
                 GAACACCA 
                   
               
               
                   
                 sa 
                   
                 asc 
                   
                   
                   
               
               
                   
               
               
                 AD-1230972 
                 uscsua(Chd)aGfu 
                 479 
                 VPusAfsacuUfuuc 
                 656 
                 CAUUUACAGUACUGG 
                 833 
               
               
                   
                 AfCfUfggaaaagus 
                   
                 caguAfcUfguagas 
                   
                 AAAAGUUU 
                   
               
               
                   
                 usa 
                   
                 usg 
                   
                   
                   
               
               
                   
               
               
                 AD-1230973 
                 csusacag(Uhd)aC 
                 480 
                 VPusAfsaacUfuuu 
                 657 
                 AUUUACAGUACUGGA 
                 834 
               
               
                   
                 fUfGfgaaaaguusu 
                   
                 ccagUfaCfuguags 
                   
                 AAAGUUUG 
                   
               
               
                   
                 sa 
                   
                 asu 
                   
                   
                   
               
               
                   
               
               
                 AD-1230977 
                 ususuuggAfcAfAf 
                 481 
                 VPusGfsaguAfcag 
                 658 
                 GAUUUUGGACAAAUC 
                 835 
               
               
                   
                 Afucug(Uhd)acus 
                   
                 auuuGfuCfcaaaas 
                   
                 UGUACCCU 
                   
               
               
                   
                 csa 
                   
                 usc 
                   
                   
                   
               
               
                   
               
               
                 AD-1230978 
                 ususugga(Chd)aA 
                 482 
                 VPusAfsgagUfaca 
                 659 
                 AUUUUGGACAAAUCU 
                 836 
               
               
                   
                 fAfUfcuguacucsu 
                   
                 gauuUfgUfccaaas 
                   
                 GUACCCUU 
                   
               
               
                   
                 sa 
                   
                 asu 
                   
                   
                   
               
               
                   
               
               
                 AD-1230979 
                 gsascaaa(Uhd)cU 
                 483 
                 VPusUfscaaAfgag 
                 660 
                 UGGACAAAUCUGUAC 
                 837 
               
               
                   
                 fGfUfacucuuugsa 
                   
                 uacaGfaUfuugucs 
                   
                 CCUUUGAC 
                   
               
               
                   
                 sa 
                   
                 csa 
                   
                   
                   
               
               
                   
               
               
                 AD-1230980 
                 csusgua(Chd)Ufc 
                 484 
                 VPusGfsgaaCfugu 
                 661 
                 AUCUGUACCCUUUGA 
                 838 
               
               
                   
                 UfUfUfgacaguucs 
                   
                 caaaGfaGfuacags 
                   
                 CUGUUCCC 
                   
               
               
                   
                 csa 
                   
                 asu 
                   
                   
                   
               
               
                   
               
               
                 AD-1230981 
                 usascuc(Uhd)Ufu 
                 485 
                 VPusAfsaggGfaac 
                 662 
                 UGUACCCUUUGACUG 
                 839 
               
               
                   
                 GfAfCfaguucccus 
                   
                 ugucAfaAfgaguas 
                   
                 UUCCCUUU 
                   
               
               
                   
                 usa 
                   
                 csa 
                   
                   
                   
               
               
                   
               
               
                 AD-1230982 
                 csuscuu(Uhd)Gfa 
                 486 
                 VPusCfsaaaGfgga 
                 663 
                 UACCCUUUGACUGUU 
                 840 
               
               
                   
                 CfAfGfuucccuuus 
                   
                 acugUfcAfaagags 
                   
                 CCCUUUGC 
                   
               
               
                   
                 gsa 
                   
                 usa 
                   
                   
                   
               
               
                   
               
               
                 AD-1230983 
                 gsusucu(Uhd)Ufg 
                 487 
                 VPusGfsaccAfaca 
                 664 
                 AAAUUCUUUGUUUCU 
                 841 
               
               
                   
                 UfAfUfcuguuggus 
                   
                 gauaCfaAfagaacs 
                   
                 GUUGGCCU 
                   
               
               
                   
                 csa 
                   
                 usu 
                   
                   
                   
               
               
                   
               
               
                 AD-1230984 
                 ususgua(Uhd)Cfu 
                 488 
                 VPusAfsggaAfgac 
                 665 
                 CUUUGUUUCUGUUGG 
                 842 
               
               
                   
                 GfUfUfggucuuccs 
                   
                 caacAfgAfuacaas 
                   
                 CCUUCCUC 
                   
               
               
                   
                 usa 
                   
                 asg 
                   
                   
                   
               
               
                   
               
               
                 AD-1230986 
                 csusaaua(Uhd)gA 
                 489 
                 VPusGfsaauCfcuu 
                 666 
                 UCCUCAUAUGACUCA 
                 843 
               
               
                   
                 fCfUfcaaggauusc 
                   
                 gaguCfaUfauuags 
                   
                 AGGAUUCU 
                   
               
               
                   
                 sa 
                   
                 gsa 
                   
                   
                   
               
               
                   
               
               
                 AD-1230987 
                 usasaua(Uhd)Gfa 
                 490 
                 VPusAfsgaaUfccu 
                 667 
                 CCUCAUAUGACUCAA 
                 844 
               
               
                   
                 CfUfCfaaggauucs 
                   
                 ugagUfcAfuauuas 
                   
                 GGAUUCUG 
                   
               
               
                   
                 usa 
                   
                 gsg 
                   
                   
                   
               
               
                   
               
               
                 AD-1230988 
                 uscsaaggAf(Uhd) 
                 491 
                 VPusAfsauuUfucc 
                 668 
                 ACUCAAGGAUUCUGG 
                 845 
               
               
                   
                 UfCfUfgggaaaaus 
                   
                 cagaAfuCfcuugas 
                   
                 GCAAACUC 
                   
               
               
                   
                 usa 
                   
                 gsu 
                   
                   
                   
               
               
                   
               
               
                 AD-1230989 
                 csasagga(Uhd)uC 
                 492 
                 VPusGfsaauUfuuc 
                 669 
                 CUCAAGGAUUCUGGG 
                 846 
               
               
                   
                 fUfGfggaaaauusc 
                   
                 ccagAfaUfccuugs 
                   
                 CAAACUCU 
                   
               
               
                   
                 sa 
                   
                 asg 
                   
                   
                   
               
               
                   
               
               
                 AD-1230990 
                 asgsgau(Uhd)Cfu 
                 493 
                 VPusUfsggaAfuuu 
                 670 
                 CAAGGAUUCUGGGCA 
                 847 
               
               
                   
                 GfGfGfaaaauuccs 
                   
                 ucccAfgAfauccus 
                   
                 AACUCUAU 
                   
               
               
                   
                 asa 
                   
                 usg 
                   
                   
                   
               
               
                   
               
               
                 AD-1230992 
                 gsgsgaaa(Uhd)cA 
                 494 
                 VPusAfsgaaAfgug 
                 671 
                 UGGAGAAAUCAUGUC 
                 848 
               
               
                   
                 fUfGfucacuuucsu 
                   
                 acauGfaUfuucccs 
                   
                 ACUUUCUG 
                   
               
               
                   
                 sa 
                   
                 csa 
                   
                   
                   
               
               
                   
               
               
                 AD-1230993 
                 usasaaa(Uhd)Cfc 
                 495 
                 VPusCfsagaAfgac 
                 672 
                 UCUGAAAUCCAUUGG 
                 849 
               
               
                   
                 AfUfUfggucuucus 
                   
                 caauGfgAfuuuuas 
                   
                 UCUUCUGC 
                   
               
               
                   
                 gsa 
                   
                 asa 
                   
                   
                   
               
               
                   
               
               
                 AD-1230994 
                 asuscca(Uhd)Ufg 
                 496 
                 VPusGfsugaCfaga 
                 673 
                 AAAUCCAUUGGUCUU 
                 850 
               
               
                   
                 GfUfCfuucugucas 
                   
                 agacCfaAfuggaus 
                   
                 CUGCCAUC 
                   
               
               
                   
                 csa 
                   
                 usu 
                   
                   
                   
               
               
                   
               
               
                 AD-1230996 
                 asusguu(Uhd)Cfu 
                 497 
                 VPusUfsgagUfaau 
                 674 
                 CCAUGUUUCUAAUGA 
                 851 
               
               
                   
                 AfAfUfgauuacucs 
                   
                 cauuAfgAfaacaus 
                   
                 UUACUCAU 
                   
               
               
                   
                 asa 
                   
                 gsg 
                   
                   
                   
               
               
                   
               
               
                 AD-1230997 
                 csasguu(Uhd)Cfa 
                 498 
                 VPusCfsaaaGfugc 
                 675 
                 UCCAGUUUCAAGAAG 
                 852 
               
               
                   
                 AfGfAfagcacuuus 
                   
                 uucuUfgAfaacugs 
                   
                 CUCUUUGU 
                   
               
               
                   
                 gsa 
                   
                 gsa 
                   
                   
                   
               
               
                   
               
               
                 AD-1230998 
                 csusgca(Chd)Afa 
                 499 
                 VPusGfsagaUfguc 
                 676 
                 CUCUGCACAAAUGUG 
                 853 
               
               
                   
                 AfUfGfugacaucus 
                   
                 acauUfuGfugcags 
                   
                 ACAUCUCA 
                   
               
               
                   
                 csa 
                   
                 asg 
                   
                   
                   
               
               
                   
               
               
                 AD-1230999 
                 csascuug(Chd)uU 
                 500 
                 VPusAfsuaaUfucc 
                 677 
                 UUCACUUGCUUCUUG 
                 854 
               
               
                   
                 fCfUfuggaauuasu 
                   
                 aagaAfgCfaagugs 
                   
                 GAAUUAUA 
                   
               
               
                   
                 sa 
                   
                 asa 
                   
                   
                   
               
               
                   
               
               
                 AD-1231001 
                 ususugaaAfcCfAf 
                 501 
                 VPusAfsggaGfauu 
                 678 
                 GAUUUGAAACCAAGA 
                 855 
               
               
                   
                 Afgaau(Chd)uccs 
                   
                 cuugGfuUfucaaas 
                   
                 GUCUCCUU 
                   
               
               
                   
                 usa 
                   
                 usu 
                   
                   
                   
               
               
                   
               
               
                 AD-1231002 
                 asuscau(Uhd)Cfc 
                 502 
                 VPusAfscuuCfagu 
                 679 
                 AUGUCAUUCCUAGAA 
                 856 
               
               
                   
                 UfAfGfaacugaags 
                   
                 ucuaGfgAfaugaus 
                   
                 GUGAAGUU 
                   
               
               
                   
                 usa 
                   
                 asu 
                   
                   
                   
               
               
                   
               
               
                 AD-1231003 
                 csasuuc(Chd)Ufa 
                 503 
                 VPusCfsaacUfuca 
                 680 
                 GUCAUUCCUAGAAGU 
                 857 
               
               
                   
                 GfAfAfcugaaguus 
                   
                 guucUfaGfgaaugs 
                   
                 GAAGUUGA 
                   
               
               
                   
                 gsa 
                   
                 asu 
                   
                   
                   
               
               
                   
               
               
                 AD-1231004 
                 ususcc(Uhd)aGfa 
                 504 
                 VPusUfsucaAfcuu 
                 681 
                 CAUUCCUAGAAGUGA 
                 858 
               
               
                   
                 AfCfUfgaaguugas 
                   
                 caguUfcUfaggaas 
                   
                 AGUUGAAG 
                   
               
               
                   
                 asa 
                   
                 usg 
                   
                   
                   
               
               
                   
               
               
                 AD-1231009 
                 ususcau(Uhd)Gfa 
                 505 
                 VPusCfsugaAfagc 
                 682 
                 UCUUCACUGACAUUG 
                 859 
               
               
                   
                 CfAfUfugcuuucas 
                   
                 aaugUfcAfaugaas 
                   
                 CUUUCAGU 
                   
               
               
                   
                 gsa 
                   
                 gsa 
                   
                   
                   
               
               
                   
               
               
                 AD-1231010 
                 csasuuga(Chd)aU 
                 506 
                 VPusUfsacuGfaaa 
                 683 
                 UUCACUGACAUUGCU 
                 860 
               
               
                   
                 fUfGfcuuucagusa 
                   
                 gcaaUfgUfcaaugs 
                   
                 UUCAGUAU 
                   
               
               
                   
                 sa 
                   
                 asa 
                   
                   
                   
               
               
                   
               
               
                 AD-1231011 
                 ascsauug(Chd)uU 
                 507 
                 VPusAfsuaaAfuac 
                 684 
                 UGACAUUGCUUUCAG 
                 861 
               
               
                   
                 fUfCfaguauuuasu 
                   
                 ugaaAfgCfaaugus 
                   
                 UAUUUAUU 
                   
               
               
                   
                 sa 
                   
                 csa 
                   
                   
                   
               
               
                   
               
               
                 AD-1231012 
                 csasgua(Uhd)Ufu 
                 508 
                 VPusAfsgagAfcag 
                 685 
                 UUCAGUAUUUAUUUC 
                 862 
               
               
                   
                 AfUfUfucugucucs 
                   
                 aaauAfaAfuacugs 
                   
                 UGCCUAAG 
                   
               
               
                   
                 usa 
                   
                 asa 
                   
                   
                   
               
               
                   
               
               
                 AD-1231022 
                 usascaagAfaAfUf 
                 509 
                 VPusGfsagaUfucu 
                 686 
                 ACUACAAGAAAUCCA 
                 863 
               
               
                   
                 Ufcagaa(Uhd)cus 
                   
                 gaauUfuCfuuguas 
                   
                 GACUCCGA 
                   
               
               
                   
                 csa 
                   
                 gsu 
                   
                   
                   
               
               
                   
               
               
                 AD-1231033 
                 csasaa(Chd)gGfu 
                 510 
                 VPusGfsaauUfgug 
                 687 
                 AACAAACAGUUGAAC 
                 864 
               
               
                   
                 UfGfAfacacaauus 
                   
                 uucaAfcCfguuugs 
                   
                 ACAAUUCU 
                   
               
               
                   
                 csa 
                   
                 csu 
                   
                   
                   
               
               
                   
               
               
                 AD-1231034 
                 asascgg(Uhd)Ufg 
                 511 
                 VPusUfsagaAfuug 
                 688 
                 CAAACAGUUGAACAC 
                 865 
               
               
                   
                 AfAfCfacaauucus 
                   
                 uguuCfaAfccguus 
                   
                 AAUUCUGA 
                   
               
               
                   
                 asa 
                   
                 usg 
                   
                   
                   
               
               
                   
               
               
                 AD-1231035 
                 ascsacaa(Uhd)uC 
                 512 
                 VPusCfsauuGfuau 
                 689 
                 GAACACAAUUCUGAA 
                 866 
               
               
                   
                 fUfAfaauacaausg 
                   
                 uuagAfaUfugugus 
                   
                 CACCAUGA 
                   
               
               
                   
                 sa 
                   
                 usc 
                   
                   
                   
               
               
                   
               
               
                 AD-1231036 
                 asuscua(Chd)Afg 
                 513 
                 VPusAfscuuUfucc 
                 690 
                 CCAUUUACAGUACUG 
                 867 
               
               
                   
                 UfAfCfuggaaaags 
                   
                 aguaCfuGfuagaus 
                   
                 GAAAAGUU 
                   
               
               
                   
                 usa 
                   
                 gsg 
                   
                   
                   
               
               
                   
               
               
                 AD-1231037 
                 usascag(Uhd)Afc 
                 514 
                 VPusCfsaaaCfuuu 
                 691 
                 UUUACAGUACUGGAA 
                 868 
               
               
                   
                 UfGfGfaaaaguuus 
                   
                 uccaGfuAfcuguas 
                   
                 AAGUUUGC 
                   
               
               
                   
                 gsa 
                   
                 gsa 
                   
                   
                   
               
               
                   
               
               
                 AD-1231038 
                 ascsagua(Chd)uG 
                 515 
                 VPusAfscaaAfcuu 
                 692 
                 UUACAGUACUGGAAA 
                 869 
               
               
                   
                 fGfAfaaaguuugsu 
                   
                 uuccAfgUfacugus 
                   
                 AGUUUGCA 
                   
               
               
                   
                 sa 
                   
                 asg 
                   
                   
                   
               
               
                   
               
               
                 AD-1231039 
                 usascuggAfaAfAf 
                 516 
                 VPusGfsguuAfcaa 
                 693 
                 AGUACUGGAAAAGUU 
                 870 
               
               
                   
                 Gfuuug(Uhd)aacs 
                   
                 acuuUfuCfcaguas 
                   
                 UGCAACCC 
                   
               
               
                   
                 csa 
                   
                 csu 
                   
                   
                   
               
               
                   
               
               
                 AD-1231044 
                 asusuu(Uhd)gGfa 
                 517 
                 VPusAfsguaCfaga 
                 694 
                 AGAUUUUGGACAAAU 
                 871 
               
               
                   
                 CfAfAfaucuguacs 
                   
                 uuugUfcCfaaaaus 
                   
                 CUGUACCC 
                   
               
               
                   
                 usa 
                   
                 csu 
                   
                   
                   
               
               
                   
               
               
                 AD-1231045 
                 ascsaaa(Uhd)Cfu 
                 518 
                 VPusGfsucaAfaga 
                 695 
                 GGACAAAUCUGUACC 
                 872 
               
               
                   
                 GfUfAfcucuuugas 
                   
                 guacAfgAfuuugus 
                   
                 CUUUGACU 
                   
               
               
                   
                 csa 
                   
                 csc 
                   
                   
                   
               
               
                   
               
               
                 AD-1231046 
                 csasaau(Chd)Ufg 
                 519 
                 VPusUfsgucAfaag 
                 696 
                 GACAAAUCUGUACCC 
                 873 
               
               
                   
                 UfAfCfucuuugacs 
                   
                 aguaCfaGfauuugs 
                   
                 UUUGACUG 
                   
               
               
                   
                 asa 
                   
                 usc 
                   
                   
                   
               
               
                   
               
               
                 AD-1231047 
                 usasucug(Uhd)uG 
                 520 
                 VPusAfsuuaGfgaa 
                 697 
                 UGUUUCUGUUGGCCU 
                 874 
               
               
                   
                 fGfUfcuuccuaasu 
                   
                 gaccAfaCfagauas 
                   
                 UCCUCAUA 
                   
               
               
                   
                 sa 
                   
                 csa 
                   
                   
                   
               
               
                   
               
               
                 AD-1231048 
                 ususggu(Chd)Ufu 
                 521 
                 VPusAfsgucAfuau 
                 698 
                 UGUUGGCCUUCCUCA 
                 875 
               
               
                   
                 CfCfUfaauaugacs 
                   
                 uaggAfaGfaccaas 
                   
                 UAUGACUC 
                   
               
               
                   
                 usa 
                   
                 csa 
                   
                   
                   
               
               
                   
               
               
                 AD-1231049 
                 cscsuaa(Uhd)Afu 
                 522 
                 VPusAfsaucCfuug 
                 699 
                 UUCCUCAUAUGACUC 
                 876 
               
               
                   
                 GfAfCfucaaggaus 
                   
                 agucAfuAfuuaggs 
                   
                 AAGGAUUC 
                   
               
               
                   
                 usa 
                   
                 asa 
                   
                   
                   
               
               
                   
               
               
                 AD-1231050 
                 asasua(Uhd)gAfc 
                 523 
                 VPusCfsagaAfucc 
                 700 
                 CUCAUAUGACUCAAG 
                 877 
               
               
                   
                 UfCfAfaggauucus 
                   
                 uugaGfuCfauauus 
                   
                 GAUUCUGG 
                   
               
               
                   
                 gsa 
                   
                 asg 
                   
                   
                   
               
               
                   
               
               
                 AD-1231051 
                 ascscuu(Uhd)Ufc 
                 524 
                 VPusCfsauuUfcuu 
                 701 
                 CAACCUUUCCUGCUA 
                 878 
               
               
                   
                 UfGfCfuaagaaaus 
                   
                 agcaGfaAfaaggus 
                   
                 AGAAACGG 
                   
               
               
                   
                 gsa 
                   
                 usg 
                   
                   
                   
               
               
                   
               
               
                 AD-1231052 
                 asasgaca(Uhd)uU 
                 525 
                 VPusAfsacuUfguc 
                 702 
                 CCAAGACAUUUUUAA 
                 879 
               
               
                   
                 fUfUfggacaagusu 
                   
                 caaaAfaUfgucuus 
                   
                 ACAACUUU 
                   
               
               
                   
                 sa 
                   
                 gsg 
                   
                   
                   
               
               
                   
               
               
                 AD-1231053 
                 gsgsaaa(Uhd)Cfa 
                 526 
                 VPusCfsagaAfagu 
                 703 
                 GGAGAAAUCAUGUCA 
                 880 
               
               
                   
                 UfGfUfcacuuucus 
                   
                 gacaUfgAfuuuccs 
                   
                 CUUUCUGC 
                   
               
               
                   
                 gsa 
                   
                 csc 
                   
                   
                   
               
               
                   
               
               
                 AD-1231054 
                 asasauca(Uhd)gU 
                 527 
                 VPusUfsgcaGfaaa 
                 704 
                 AGAAAUCAUGUCACU 
                 881 
               
               
                   
                 fCfAfcuuucugcsa 
                   
                 gugaCfaUfgauuus 
                   
                 UUCUGCAG 
                   
               
               
                   
                 sa 
                   
                 csc 
                   
                   
                   
               
               
                   
               
               
                 AD-1231055 
                 asasuca(Uhd)Gfu 
                 528 
                 VPusCfsugcAfgaa 
                 705 
                 GAAAUCAUGUCACUU 
                 882 
               
               
                   
                 CfAfCfuuucugcas 
                   
                 agugAfcAfugauus 
                   
                 UCUGCAGC 
                   
               
               
                   
                 gsa 
                   
                 usc 
                   
                   
                   
               
               
                   
               
               
                 AD-1231058 
                 gscsauu(Uhd)Afa 
                 529 
                 VPusGfsaccAfaug 
                 706 
                 AAGCAUCUGAAAUCC 
                 883 
               
               
                   
                 AfAfUfccauuggus 
                   
                 gauuUfuAfaaugcs 
                   
                 AUUGGUCU 
                   
               
               
                   
                 csa 
                   
                 usu 
                   
                   
                   
               
               
                   
               
               
                 AD-1231065 
                 csasucu(Chd)Ufg 
                 530 
                 VPusAfsgaaAfcau 
                 707 
                 UGCAUCUCUGUUCCA 
                 884 
               
               
                   
                 UfUfCfcauguuucs 
                   
                 ggaaCfaGfagaugs 
                   
                 UGUUUCUA 
                   
               
               
                   
                 usa 
                   
                 csg 
                   
                   
                   
               
               
                   
               
               
                 AD-1231066 
                 usgsuuu(Chd)Ufa 
                 531 
                 VPusAfsugaGfuaa 
                 708 
                 CAUGUUUCUAAUGAU 
                 885 
               
               
                   
                 AfUfGfauuacucas 
                   
                 ucauUfaGfaaacas 
                   
                 UACUCAUU 
                   
               
               
                   
                 usa 
                   
                 usg 
                   
                   
                   
               
               
                   
               
               
                 AD-1231067 
                 asusgau(Uhd)Afc 
                 532 
                 VPusUfscgaAfuga 
                 709 
                 UAAUGAUUACUCAUU 
                 886 
               
               
                   
                 UfCfAfuucauucgs 
                   
                 augaGfuAfaucaus 
                   
                 CAUUCGAU 
                   
               
               
                   
                 asa 
                   
                 usa 
                   
                   
                   
               
               
                   
               
               
                 AD-1231068 
                 csasauu(Chd)Cfa 
                 533 
                 VPusGfscuuCfuug 
                 710 
                 ACCAAUUCCAGUUUC 
                 887 
               
               
                   
                 GfUfUfucaagaags 
                   
                 aaacUfgGfaauugs 
                   
                 AAGAAGCU 
                   
               
               
                   
                 csa 
                   
                 gsu 
                   
                   
                   
               
               
                   
               
               
                 AD-1231069 
                 gsusuu(Chd)aAfg 
                 534 
                 VPusGfsacaAfagu 
                 711 
                 CAGUUUCAAGAAGCU 
                 888 
               
               
                   
                 AfAfGfcacuuugus 
                   
                 gcuuCfuUfgaaacs 
                   
                 CUUUGUCA 
                   
               
               
                   
                 csa 
                   
                 usg 
                   
                   
                   
               
               
                   
               
               
                 AD-1231070 
                 ususucaaGfaAfGf 
                 535 
                 VPusUfsgacAfaag 
                 712 
                 AGUUUCAAGAAGCUC 
                 889 
               
               
                   
                 Cfacuu(Uhd)gucs 
                   
                 ugcuUfcUfugaaas 
                   
                 UUUGUCAA 
                   
               
               
                   
                 asa 
                   
                 csu 
                   
                   
                   
               
               
                   
               
               
                 AD-1231071 
                 csusuug(Uhd)Cfa 
                 536 
                 VPusUfsguuUfagc 
                 713 
                 CUCUUUGUCAAGCAG 
                 890 
               
               
                   
                 AfGfCfagcuaaacs 
                   
                 ugcuUfgAfcaaags 
                   
                 CUAAGUAU 
                   
               
               
                   
                 asa 
                   
                 usg 
                   
                   
                   
               
               
                   
               
               
                 AD-1231072 
                 ususcua(Uhd)Cfa 
                 537 
                 VPusGfscaaGfuga 
                 714 
                 UGUCUUAUCAAAGUU 
                 891 
               
               
                   
                 AfAfGfuucacuugs 
                   
                 acuuUfgAfuagaas 
                   
                 CACUUGCU 
                   
               
               
                   
                 csa 
                   
                 csa 
                   
                   
                   
               
               
                   
               
               
                 AD-1231073 
                 asasguu(Chd)Afc 
                 538 
                 VPusUfsccaAfgaa 
                 715 
                 CAAAGUUCACUUGCU 
                 892 
               
               
                   
                 UfUfGfcuucuuggs 
                   
                 gcaaGfuGfaacuus 
                   
                 UCUUGGAA 
                   
               
               
                   
                 asa 
                   
                 usg 
                   
                   
                   
               
               
                   
               
               
                 AD-1231075 
                 ususcac(Uhd)Ufg 
                 539 
                 VPusAfsauuCfcaa 
                 716 
                 AGUUCACUUGCUUCU 
                 893 
               
               
                   
                 CfUfUfcuuggaaus 
                   
                 gaagCfaAfgugaas 
                   
                 UGGAAUUA 
                   
               
               
                   
                 usa 
                   
                 csu 
                   
                   
                   
               
               
                   
               
               
                 AD-1231076 
                 ascsuug(Chd)Ufu 
                 540 
                 VPusUfsauaAfuuc 
                 717 
                 UCACUUGCUUCUUGG 
                 894 
               
               
                   
                 CfUfUfggaauuaus 
                   
                 caagAfaGfcaagus 
                   
                 AAUUAUAA 
                   
               
               
                   
                 asa 
                   
                 gsa 
                   
                   
                   
               
               
                   
               
               
                 AD-1231077 
                 gscsuuc(Uhd)Ufg 
                 541 
                 VPusGfsuguUfaua 
                 718 
                 UUGCUUCUUGGAAUU 
                 895 
               
               
                   
                 GfAfAfuuauaacas 
                   
                 auucCfaAfgaagcs 
                   
                 AUAAUACU 
                   
               
               
                   
                 csa 
                   
                 asa 
                   
                   
                   
               
               
                   
               
               
                 AD-1231080 
                 csusucu(Uhd)Gfg 
                 542 
                 VPusGfsgugUfuau 
                 719 
                 UGCUUCUUGGAAUUA 
                 896 
               
               
                   
                 AfAfUfuauaacacs 
                   
                 aauuCfcAfagaags 
                   
                 UAAUACUA 
                   
               
               
                   
                 csa 
                   
                 csa 
                   
                   
                   
               
               
                   
               
               
                 AD-1231081 
                 csasaag(Chd)Afu 
                 543 
                 VPusAfsuccUfcac 
                 720 
                 ACCAAAGCAUUAAAG 
                 897 
               
               
                   
                 CfAfAfagugaggas 
                   
                 uuugAfuGfcuuugs 
                   
                 UGAGGAUA 
                   
               
               
                   
                 usa 
                   
                 gsu 
                   
                   
                   
               
               
                   
               
               
                 AD-1231082 
                 asgscuc(Uhd)Ufg 
                 544 
                 VPusAfsugcUfuua 
                 721 
                 UCAGCUCUUGGAGCU 
                 898 
               
               
                   
                 GfAfGfauaaagcas 
                   
                 ucucCfaAfgagcus 
                   
                 AAUGCAUA 
                   
               
               
                   
                 usa 
                   
                 gsa 
                   
                   
                   
               
               
                   
               
               
                 AD-1231083 
                 asusgua(Chd)Cfu 
                 545 
                 VPusGfsaugAfucg 
                 722 
                 AAAUGUUCCUGUUCC 
                 899 
               
               
                   
                 GfUfUfccgaucaus 
                   
                 gaacAfgGfuacaus 
                   
                 GAUCAUCU 
                   
               
               
                   
                 csa 
                   
                 usu 
                   
                   
                   
               
               
                   
               
               
                 AD-1231085 
                 ususac(Uhd)gAfa 
                 546 
                 VPusUfsuggAfcau 
                 723 
                 CAUUACUGAAGAAAA 
                 900 
               
               
                   
                 GfAfGfaauguccas 
                   
                 ucucUfuCfaguaas 
                   
                 UGCCCAAA 
                   
               
               
                   
                 asa 
                   
                 usa 
                   
                   
                   
               
               
                   
               
               
                 AD-1231087 
                 asusucc(Uhd)Afg 
                 547 
                 VPusUfscaaCfuuc 
                 724 
                 UCAUUCCUAGAAGUG 
                 901 
               
               
                   
                 AfAfCfugaaguugs 
                   
                 aguuCfuAfggaaus 
                   
                 AAGUUGAA 
                   
               
               
                   
                 asa 
                   
                 gsa 
                   
                   
                   
               
               
                   
               
               
                 AD-1231091 
                 uscscaggAfuUfCf 
                 548 
                 VPusCfsaguGfuuu 
                 725 
                 AAUGCAGGAUUCCAA 
                 902 
               
               
                   
                 Cfaaaa(Chd)acus 
                   
                 uggaAfuCfcuggas 
                   
                 AACAGUGA 
                   
               
               
                   
                 gsa 
                   
                 usu 
                   
                   
                   
               
               
                   
               
               
                 AD-1231092 
                 escsaaaa(Chd)aC 
                 549 
                 VPusGfsaacAfuca 
                 726 
                 UUCCAAAACAGUGAU 
                 903 
               
               
                   
                 fUfGfaugauguusc 
                   
                 ucagUfgUfuuuggs 
                   
                 GAUGCUCA 
                   
               
               
                   
                 sa 
                   
                 asa 
                   
                   
                   
               
               
                   
               
               
                 AD-1231093 
                 gsasccu(Chd)Cfu 
                 550 
                 VPusGfsauuUfuuc 
                 727 
                 CAGACUUCCUUUUAG 
                 904 
               
               
                   
                 UfUfUfagaaaaaus 
                   
                 uaaaAfgGfaggucs 
                   
                 CAAAGCAC 
                   
               
               
                   
                 csa 
                   
                 usg 
                   
                   
                   
               
               
                   
               
               
                 AD-1231098 
                 uscsau(Uhd)gAfc 
                 551 
                 VPusAfscugAfaag 
                 728 
                 CUUCACUGACAUUGC 
                 905 
               
               
                   
                 AfUfUfgcuuucags 
                   
                 caauGfuCfaaugas 
                   
                 UUUCAGUA 
                   
               
               
                   
                 usa 
                   
                 asg 
                   
                   
                   
               
               
                   
               
               
                 AD-1231099 
                 asusuga(Chd)Afu 
                 552 
                 VPusAfsuacUfgaa 
                 729 
                 UCACUGACAUUGCUU 
                 906 
               
               
                   
                 UfGfCfuuucaguas 
                   
                 agcaAfuGfucaaus 
                   
                 UCAGUAUU 
                   
               
               
                   
                 usa 
                   
                 gsa 
                   
                   
                   
               
               
                   
               
               
                 AD-1231100 
                 csasuug(Chd)Ufu 
                 553 
                 VPusAfsauaAfaua 
                 730 
                 GACAUUGCUUUCAGU 
                 907 
               
               
                   
                 UfCfAfguauuuaus 
                   
                 cugaAfaGfcaaugs 
                   
                 AUUUAUUU 
                   
               
               
                   
                 usa 
                   
                 usc 
                   
                   
                   
               
               
                   
               
               
                 AD-1231101 
                 ususgcu(Uhd)Ufc 
                 554 
                 VPusGfsaaaUfaaa 
                 731 
                 CAUUGCUUUCAGUAU 
                 908 
               
               
                   
                 AfGfUfauuuauuus 
                   
                 uacuGfaAfagcaas 
                   
                 UUAUUUCU 
                   
               
               
                   
                 csa 
                   
                 usg 
                   
                   
                   
               
               
                   
               
               
                 AD-1231102 
                 csusuu(Chd)aGfu 
                 555 
                 VPusAfscagAfaau 
                 732 
                 UGCUUUCAGUAUUUA 
                 909 
               
               
                   
                 AfUfUfuauuucugs 
                   
                 aaauAfcUfgaaags 
                   
                 UUUCUGCC 
                   
               
               
                   
                 usa 
                   
                 csa 
                   
                   
                   
               
               
                   
               
               
                 AD-1231103 
                 ususucag(Uhd)aU 
                 556 
                 VPusGfsacaGfaaa 
                 733 
                 GCUUUCAGUAUUUAU 
                 910 
               
               
                   
                 fUfUfauuucugusc 
                   
                 uaaaUfaCfugaaas 
                   
                 UUCUGCCU 
                   
               
               
                   
                 sa 
                   
                 gsc 
                   
                   
                   
               
               
                   
               
               
                 AD-1231104 
                 ususcag(Uhd)Afu 
                 557 
                 VPusAfsgacAfgaa 
                 734 
                 CUUUCAGUAUUUAUU 
                 911 
               
               
                   
                 UfUfAfuuucugucs 
                   
                 auaaAfuAfcugaas 
                   
                 UCUGCCUA 
                   
               
               
                   
                 usa 
                   
                 asg 
                   
                   
                   
               
               
                   
               
               
                 AD-1231108 
                 asusuuga(Chd)uU 
                 558 
                 VPusAfsaacAfgaa 
                 735 
                 GGAUUUGACAUCUCU 
                 912 
               
               
                   
                 fCfUfguucuguusu 
                   
                 cagaAfgUfcaaaus 
                   
                 UCUGUUUA 
                   
               
               
                   
                 sa 
                   
                 csc 
                   
                   
                   
               
               
                   
               
               
                 AD-1231118 
                 usgscc(Uhd)gAfu 
                 559 
                 VPusAfsaauGfagu 
                 736 
                 CAUGCCUGUCAGAAA 
                 913 
               
               
                   
                 AfGfAfaacucauus 
                   
                 uucuAfuCfaggcas 
                   
                 CUACUUCC 
                   
               
               
                   
                 usa 
                   
                 usg 
               
               
                   
               
            
           
         
       
     
     
       
         
           
               
             
               
                 TABLE 4 
               
             
            
               
                   
               
               
                 Unmodified Sense and Antisense Strand Cynomolgus Monkey ACE2 dsRNA Sequences 
               
            
           
           
               
               
               
               
               
               
               
            
               
                   
                   
                 SEQ 
                 Range in 
                   
                 SEQ 
                 Range in 
               
               
                   
                   
                 ID 
                 XM_0055930 
                   
                 ID 
                 XM_0055930 
               
               
                 Duplex Name 
                 Sense Sequence 5′ to 3′ 
                 NO: 
                 37.2 
                 Antisense Sequence 5′ to 3′ 
                 NO: 
                 37.2 
               
               
                   
               
            
           
           
               
               
               
               
               
               
               
            
               
                 AD-1230824 
                 CUAAGCAUUUAAAAUCCAUUA 
                 914 
                 1544-1564 
                 UAAUGGAUUUUAAAUGCUUAGGU 
                 1036 
                 1542-1564 
               
               
                   
               
               
                 AD-1230830 
                 CAUCUUCAUUGACAUUGCUUA 
                 915 
                 2875-2895 
                 UAAGCAAUGUCAAUGAAGAUGCU 
                 1037 
                 2873-2895 
               
               
                   
               
               
                 AD-1230834 
                 CUGGGAAAAUUCCAUGCUAAA 
                 916 
                 1281-1301 
                 UUUAGCAUGGAAUUUUCCCAGAA 
                 1038 
                 1279-1301 
               
               
                   
               
               
                 AD-1230835 
                 CCUAAGCAUUUAAAAUCCAUA 
                 917 
                 1543-1563 
                 UAUGGAUUUUAAAUGCUUAGGUG 
                 1039 
                 1541-1563 
               
               
                   
               
               
                 AD-1230840 
                 GAAGACCUGUUCUAUCAAAGA 
                 918 
                 409-429 
                 UCUUUGAUAGAACAGGUCUUCGG 
                 1040 
                 407-429 
               
               
                   
               
               
                 AD-1230850 
                 AAAUGUGACAUCUCAAACUCA 
                 919 
                 1921-1941 
                 UGAGUUUGAGAUGUCACAUUUGU 
                 1041 
                 1919-1941 
               
               
                   
               
               
                 AD-1230860 
                 UAAAUGUCUGUUGAAUUUCUA 
                 920 
                 3018-3038 
                 UAGAAAUUCAACAGACAUUUACA 
                 1042 
                 3016-3038 
               
               
                   
               
               
                 AD-1230861 
                 GCUCACUUUCAUUUAAUCCAA 
                 921 
                 3164-3184 
                 UUGGAUUAAAUGAAAGUGAGCUA 
                 1043 
                 3162-3184 
               
               
                   
               
               
                 AD-1230862 
                 CACUUUCAUUUAAUCCAUUGA 
                 922 
                 3167-3187 
                 UCAAUGGAUUAAAUGAAAGUGAG 
                 1044 
                 3165-3187 
               
               
                   
               
               
                 AD-1230863 
                 AUUGCUUUUUCACUUCCAAGA 
                 923 
                 3326-3346 
                 UCUUGGAAGUGAAAAAGCAAUGU 
                 1045 
                 3324-3346 
               
               
                   
               
               
                 AD-1230864 
                 CAGAAUCUCACAGUCAAGCUA 
                 924 
                 565-585 
                 UAGCUUGACUGUGAGAUUCUGAA 
                 1046 
                 563-585 
               
               
                   
               
               
                 AD-1230865 
                 AGAAUCUCACAGUCAAGCUUA 
                 925 
                 566-586 
                 UAAGCUUGACUGUGAGAUUCUGA 
                 1047 
                 564-586 
               
               
                   
               
               
                 AD-1230866 
                 CAGUCUCUUAAAUCUUUUGUA 
                 926 
                 3500-3520 
                 UACAAAAGAUUUAAGAGACUGGG 
                 1048 
                 3498-3520 
               
               
                   
               
               
                 AD-1230871 
                 CUUCCUAAUAUGACUCAAGGA 
                 927 
                 1258-1278 
                 UCCUUGAGUCAUAUUAGGAAGAC 
                 1049 
                 1256-1278 
               
               
                   
               
               
                 AD-1230872 
                 UGGGAAAAUUCCAUGCUAACA 
                 928 
                 1282-1302 
                 UGUUAGCAUGGAAUUUUCCCAGA 
                 1050 
                 1280-1302 
               
               
                   
               
               
                 AD-1230893 
                 CUCAAACUCUACAGAAGCUGA 
                 929 
                 1932-1952 
                 UCAGCUUCUGUAGAGUUUGAGAU 
                 1051 
                 1930-1952 
               
               
                   
               
               
                 AD-1230898 
                 CCAAGAAUCUCCUUUAAUUUA 
                 930 
                 2329-2349 
                 UAAAUUAAAGGAGAUUCUUGGUU 
                 1052 
                 2327-2349 
               
               
                   
               
               
                 AD-1230899 
                 CAAGAAUCUCCUUUAAUUUCA 
                 931 
                 2330-2350 
                 UGAAAUUAAAGGAGAUUCUUGGU 
                 1053 
                 2328-2350 
               
               
                   
               
               
                 AD-1230900 
                 AAGAAUCUCCUUUAAUUUCUA 
                 932 
                 2331-2351 
                 UAGAAAUUAAAGGAGAUUCUUGG 
                 1054 
                 2329-2351 
               
               
                   
               
               
                 AD-1230901 
                 CUGCACCUAAAAAUGUGUCUA 
                 933 
                 2357-2377 
                 UAGACACAUUUUUAGGUGCAGUG 
                 1055 
                 2355-2377 
               
               
                   
               
               
                 AD-1230903 
                 UCACUUUCAUUUAAUCCAUUA 
                 934 
                 3166-3186 
                 UAAUGGAUUAAAUGAAAGUGAGC 
                 1056 
                 3164-3186 
               
               
                   
               
               
                 AD-1230905 
                 AAUUCAGAAUCUCACAGUCAA 
                 935 
                 561-581 
                 UUGACUGUGAGAUUCUGAAUUUC 
                 1057 
                 559-581 
               
               
                   
               
               
                 AD-1230906 
                 AUUCAGAAUCUCACAGUCAAA 
                 936 
                 562-582 
                 UUUGACUGUGAGAUUCUGAAUUU 
                 1058 
                 560-582 
               
               
                   
               
               
                 AD-1230907 
                 UUCAGAAUCUCACAGUCAAGA 
                 937 
                 563-583 
                 UCUUGACUGUGAGAUUCUGAAUU 
                 1059 
                 561-583 
               
               
                   
               
               
                 AD-1230908 
                 GCCUGAUAGAAACUCAUUUCA 
                 938 
                 3414-3434 
                 UGAAAUGAGUUUCUAUCAGGCAU 
                 1060 
                 3412-3434 
               
               
                   
               
               
                 AD-1230909 
                 CCAGGUUUGAAUGAAAUAAUA 
                 939 
                 736-756 
                 UAUUAUUUCAUUCAAACCUGGUU 
                 1061 
                 734-756 
               
               
                   
               
               
                 AD-1230917 
                 UCUGGGAAAAUUCCAUGCUAA 
                 940 
                 1280-1300 
                 UUAGCAUGGAAUUUUCCCAGAAU 
                 1062 
                 1278-1300 
               
               
                   
               
               
                 AD-1230918 
                 CACAACCUUUUCUGCUAAGAA 
                 941 
                 1460-1480 
                 UUCUUAGCAGAAAAGGUUGUGCA 
                 1063 
                 1458-1480 
               
               
                   
               
               
                 AD-1230919 
                 CAACCUUUUCUGCUAAGAAAA 
                 942 
                 1462-1482 
                 UUUUCUUAGCAGAAAAGGUUGUG 
                 1064 
                 1460-1482 
               
               
                   
               
               
                 AD-1230924 
                 GAAACAGAAAUAAACUUCCUA 
                 943 
                 1597-1617 
                 UAGGAAGUUUAUUUCUGUUUCAU 
                 1065 
                 1595-1617 
               
               
                   
               
               
                 AD-1230925 
                 UCAAACAAGCACUCACGAUUA 
                 944 
                 1619-1639 
                 UAAUCGUGAGUGCUUGUUUGAGC 
                 1066 
                 1617-1639 
               
               
                   
               
               
                 AD-1230926 
                 UCUGCCAUUUACUUACAUGUA 
                 945 
                 1647-1667 
                 UACAUGUAAGUAAAUGGCAGAGU 
                 1067 
                 1645-1667 
               
               
                   
               
               
                 AD-1230927 
                 CUGCCAUUUACUUACAUGUUA 
                 946 
                 1648-1668 
                 UAACAUGUAAGUAAAUGGCAGAG 
                 1068 
                 1646-1668 
               
               
                   
               
               
                 AD-1230929 
                 CGAAGACCUGUUCUAUCAAAA 
                 947 
                 408-428 
                 UUUUGAUAGAACAGGUCUUCGGC 
                 1069 
                 406-428 
               
               
                   
               
               
                 AD-1230933 
                 AAGACCUGUUCUAUCAAAGUA 
                 948 
                 410-430 
                 UACUUUGAUAGAACAGGUCUUCG 
                 1070 
                 408-430 
               
               
                   
               
               
                 AD-1230943 
                 AUCAAUGAUGCUUUCCGUCUA 
                 949 
                 2431-2451 
                 UAGACGGAAAGCAUCAUUGAUAC 
                 1071 
                 2429-2451 
               
               
                   
               
               
                 AD-1230944 
                 AAUGUCCAAAACAUGAAUAAA 
                 950 
                 472-492 
                 UUUAUUCAUGUUUUGGACAUUCU 
                 1072 
                 470-492 
               
               
                   
               
               
                 AD-1230945 
                 AUGUCCAAAACAUGAAUAAUA 
                 951 
                 473-493 
                 UAUUAUUCAUGUUUUGGACAUUC 
                 1073 
                 471-493 
               
               
                   
               
               
                 AD-1230952 
                 UCUGUCUCUGGAUUUGACUUA 
                 952 
                 2907-2927 
                 UAAGUCAAAUCCAGAGACAGAAA 
                 1074 
                 2905-2927 
               
               
                   
               
               
                 AD-1230953 
                 UCUGGAUUUGACUUCUGUUCA 
                 953 
                 2913-2933 
                 UGAACAGAAGUCAAAUCCAGAGA 
                 1075 
                 2911-2933 
               
               
                   
               
               
                 AD-1230954 
                 GAUUUGACUUCUGUUCUGUUA 
                 954 
                 2917-2937 
                 UAACAGAACAGAAGUCAAAUCCA 
                 1076 
                 2915-2937 
               
               
                   
               
               
                 AD-1230955 
                 CAGGGAUAAUCUAAAUGUAAA 
                 955 
                 3001-3021 
                 UUUACAUUUAGAUUAUCCCUGAA 
                 1077 
                 2999-3021 
               
               
                   
               
               
                 AD-1230956 
                 AGGGAUAAUCUAAAUGUAAAA 
                 956 
                 3002-3022 
                 UUUUACAUUUAGAUUAUCCCUGA 
                 1078 
                 3000-3022 
               
               
                   
               
               
                 AD-1230957 
                 UAAAUGUAAAUGUCUGUUGAA 
                 957 
                 3012-3032 
                 UUCAACAGACAUUUACAUUUAGA 
                 1079 
                 3010-3032 
               
               
                   
               
               
                 AD-1230958 
                 AUGUCUGUUGAAUUUCUGAAA 
                 958 
                 3021-3041 
                 UUUCAGAAAUUCAACAGACAUUU 
                 1080 
                 3019-3041 
               
               
                   
               
               
                 AD-1230959 
                 UCUGUUGAAUUUCUGAAGUUA 
                 959 
                 3024-3044 
                 UAACUUCAGAAAUUCAACAGACA 
                 1081 
                 3022-3044 
               
               
                   
               
               
                 AD-1230960 
                 CUCACUUUCAUUUAAUCCAUA 
                 960 
                 3165-3185 
                 UAUGGAUUAAAUGAAAGUGAGCU 
                 1082 
                 3163-3185 
               
               
                   
               
               
                 AD-1230961 
                 ACUUUCAUUUAAUCCAUUGUA 
                 961 
                 3168-3188 
                 UACAAUGGAUUAAAUGAAAGUGA 
                 1083 
                 3166-3188 
               
               
                   
               
               
                 AD-1230963 
                 GAAUCUCACAGUCAAGCUUCA 
                 962 
                 567-587 
                 UGAAGCUUGACUGUGAGAUUCUG 
                 1084 
                 565-587 
               
               
                   
               
               
                 AD-1230964 
                 AAUUCCAACUGUAUGUUCACA 
                 963 
                 3463-3483 
                 UGUGAACAUACAGUUGGAAUUUC 
                 1085 
                 3461-3483 
               
               
                   
               
               
                 AD-1230965 
                 AGUCUCUUAAAUCUUUUGUAA 
                 964 
                 3501-3521 
                 UUACAAAAGAUUUAAGAGACUGG 
                 1086 
                 3499-3521 
               
               
                   
               
               
                 AD-1230966 
                 GUCUCUUAAAUCUUUUGUAUA 
                 965 
                 3502-3522 
                 UAUACAAAAGAUUUAAGAGACUG 
                 1087 
                 3500-3522 
               
               
                   
               
               
                 AD-1230967 
                 CACUCAAUAAAUGCUAGAUUA 
                 966 
                 3561-3581 
                 UAAUCUAGCAUUUAUUGAGUGUC 
                 1088 
                 3559-3581 
               
               
                   
               
               
                 AD-1230968 
                 ACUCAAUAAAUGCUAGAUUUA 
                 967 
                 3562-3582 
                 UAAAUCUAGCAUUUAUUGAGUGU 
                 1089 
                 3560-3582 
               
               
                   
               
               
                 AD-1230974 
                 ACCAGGUUUGAAUGAAAUAAA 
                 968 
                 735-755 
                 UUUAUUUCAUUCAAACCUGGUUC 
                 1090 
                 733-755 
               
               
                   
               
               
                 AD-1230975 
                 ACCAUUAUAUGAACAUCUUCA 
                 969 
                 1002-1022 
                 UGAAGAUGUUCAUAUAAUGGUUU 
                 1091 
                 1000-1022 
               
               
                   
               
               
                 AD-1230976 
                 CCAUUAUAUGAACAUCUUCAA 
                 970 
                 1003-1023 
                 UUGAAGAUGUUCAUAUAAUGGUU 
                 1092 
                 1001-1023 
               
               
                   
               
               
                 AD-1230985 
                 UCUAGGGAAAGUCAUUCAGUA 
                 971 
                 254-274 
                 UACUGAAUGACUUUCCCUAGACU 
                 1093 
                 252-274 
               
               
                   
               
               
                 AD-1230991 
                 GCACAACCUUUUCUGCUAAGA 
                 972 
                 1459-1479 
                 UCUUAGCAGAAAAGGUUGUGCAG 
                 1094 
                 1457-1479 
               
               
                   
               
               
                 AD-1230995 
                 UGCCAUUUACUUACAUGUUAA 
                 973 
                 1649-1669 
                 UUAACAUGUAAGUAAAUGGCAGA 
                 1095 
                 1647-1669 
               
               
                   
               
               
                 AD-1231000 
                 GACCAGAACAAGAAUUCUUUA 
                 974 
                 2089-2109 
                 UAAAGAAUUCUUGUUCUGGUCUU 
                 1096 
                 2087-2109 
               
               
                   
               
               
                 AD-1231005 
                 UAUCAAUGAUGCUUUCCGUCA 
                 975 
                 2430-2450 
                 UGACGGAAAGCAUCAUUGAUACG 
                 1097 
                 2428-2450 
               
               
                   
               
               
                 AD-1231006 
                 UGUCCAAAACAUGAAUAAUGA 
                 976 
                 474-494 
                 UCAUUAUUCAUGUUUUGGACAUU 
                 1098 
                 472-494 
               
               
                   
               
               
                 AD-1231007 
                 CUCCUUUUAGAAAAAUCUAUA 
                 977 
                 2709-2729 
                 UAUAGAUUUUUCUAAAAGGAGGU 
                 1099 
                 2707-2729 
               
               
                   
               
               
                 AD-1231008 
                 AUUGUCCAAAGACAACAUGGA 
                 978 
                 2843-2863 
                 UCCAUGUUGUCUUUGGACAAUUU 
                 1100 
                 2841-2863 
               
               
                   
               
               
                 AD-1231013 
                 UUCUGUCUCUGGAUUUGACUA 
                 979 
                 2906-2926 
                 UAGUCAAAUCCAGAGACAGAAAU 
                 1101 
                 2904-2926 
               
               
                   
               
               
                 AD-1231014 
                 CUCUGGAUUUGACUUCUGUUA 
                 980 
                 2912-2932 
                 UAACAGAAGUCAAAUCCAGAGAC 
                 1102 
                 2910-2932 
               
               
                   
               
               
                 AD-1231015 
                 CUGGAUUUGACUUCUGUUCUA 
                 981 
                 2914-2934 
                 UAGAACAGAAGUCAAAUCCAGAG 
                 1103 
                 2912-2934 
               
               
                   
               
               
                 AD-1231016 
                 UGGAUUUGACUUCUGUUCUGA 
                 982 
                 2915-2935 
                 UCAGAACAGAAGUCAAAUCCAGA 
                 1104 
                 2913-2935 
               
               
                   
               
               
                 AD-1231017 
                 GGAUUUGACUUCUGUUCUGUA 
                 983 
                 2916-2936 
                 UACAGAACAGAAGUCAAAUCCAG 
                 1105 
                 2914-2936 
               
               
                   
               
               
                 AD-1231018 
                 GUAAAUGUCUGUUGAAUUUCA 
                 984 
                 3017-3037 
                 UGAAAUUCAACAGACAUUUACAU 
                 1106 
                 3015-3037 
               
               
                   
               
               
                 AD-1231019 
                 AAAUGUCUGUUGAAUUUCUGA 
                 985 
                 3019-3039 
                 UCAGAAAUUCAACAGACAUUUAC 
                 1107 
                 3017-3039 
               
               
                   
               
               
                 AD-1231020 
                 UGUCUGUUGAAUUUCUGAAGA 
                 986 
                 3022-3042 
                 UCUUCAGAAAUUCAACAGACAUU 
                 1108 
                 3020-3042 
               
               
                   
               
               
                 AD-1231021 
                 CUGUUGAAUUUCUGAAGUUGA 
                 987 
                 3025-3045 
                 UCAACUUCAGAAAUUCAACAGAC 
                 1109 
                 3023-3045 
               
               
                   
               
               
                 AD-1231023 
                 UCAGAAUCUCACAGUCAAGCA 
                 988 
                 564-584 
                 UGCUUGACUGUGAGAUUCUGAAU 
                 1110 
                 562-584 
               
               
                   
               
               
                 AD-1231024 
                 AUCUCACAGUCAAGCUUCAGA 
                 989 
                 569-589 
                 UCUGAAGCUUGACUGUGAGAUUC 
                 1111 
                 567-589 
               
               
                   
               
               
                 AD-1231025 
                 CUACUGUUCUCUAACUGUGGA 
                 990 
                 3433-3453 
                 UCCACAGUUAGAGAACAGUAGAA 
                 1112 
                 3431-3453 
               
               
                   
               
               
                 AD-1231026 
                 GAAUGGAAAUUCCAACUGUAA 
                 991 
                 3456-3476 
                 UUACAGUUGGAAUUUCCAUUCAC 
                 1113 
                 3454-3476 
               
               
                   
               
               
                 AD-1231027 
                 GAAAUUCCAACUGUAUGUUCA 
                 992 
                 3461-3481 
                 UGAACAUACAGUUGGAAUUUCCA 
                 1114 
                 3459-3481 
               
               
                   
               
               
                 AD-1231028 
                 CCAGUCUCUUAAAUCUUUUGA 
                 993 
                 3499-3519 
                 UCAAAAGAUUUAAGAGACUGGGU 
                 1115 
                 3497-3519 
               
               
                   
               
               
                 AD-1231029 
                 ACAAAGCAGACACUCAAUAAA 
                 994 
                 3551-3571 
                 UUUAUUGAGUGUCUGCUUUGUGC 
                 1116 
                 3549-3571 
               
               
                   
               
               
                 AD-1231030 
                 AAAGCAGACACUCAAUAAAUA 
                 995 
                 3553-3573 
                 UAUUUAUUGAGUGUCUGCUUUGU 
                 1117 
                 3551-3573 
               
               
                   
               
               
                 AD-1231031 
                 GCAGACACUCAAUAAAUGCUA 
                 996 
                 3556-3576 
                 UAGCAUUUAUUGAGUGUCUGCUU 
                 1118 
                 3554-3576 
               
               
                   
               
               
                 AD-1231032 
                 AGACACUCAAUAAAUGCUAGA 
                 997 
                 3558-3578 
                 UCUAGCAUUUAUUGAGUGUCUGC 
                 1119 
                 3556-3578 
               
               
                   
               
               
                 AD-1231040 
                 CAUACCUUUGAAGAGAUUAAA 
                 998 
                 982-1002 
                 UUUAAUCUCUUCAAAGGUAUGUU 
                 1120 
                 980-1002 
               
               
                   
               
               
                 AD-1231041 
                 AUACCUUUGAAGAGAUUAAAA 
                 999 
                 983-1003 
                 UUUUAAUCUCUUCAAAGGUAUGU 
                 1121 
                 981-1003 
               
               
                   
               
               
                 AD-1231042 
                 ACCUUUGAAGAGAUUAAACCA 
                 1000 
                 985-1005 
                 UGGUUUAAUCUCUUCAAAGGUAU 
                 1122 
                 983-1005 
               
               
                   
               
               
                 AD-1231043 
                 CAUUAUAUGAACAUCUUCAUA 
                 1001 
                 1004-1024 
                 UAUGAAGAUGUUCAUAUAAUGGU 
                 1123 
                 1002-1024 
               
               
                   
               
               
                 AD-1231056 
                 CACACCUAAGCAUUUAAAAUA 
                 1002 
                 1539-1559 
                 UAUUUUAAAUGCUUAGGUGUGGC 
                 1124 
                 1537-1559 
               
               
                   
               
               
                 AD-1231057 
                 ACCUAAGCAUUUAAAAUCCAA 
                 1003 
                 1542-1562 
                 UUGGAUUUUAAAUGCUUAGGUGU 
                 1125 
                 1540-1562 
               
               
                   
               
               
                 AD-1231059 
                 CAAUGAAACAGAAAUAAACUA 
                 1004 
                 1593-1613 
                 UAGUUUAUUUCUGUUUCAUUGUC 
                 1126 
                 1591-1613 
               
               
                   
               
               
                 AD-1231060 
                 AUGAAACAGAAAUAAACUUCA 
                 1005 
                 1595-1615 
                 UGAAGUUUAUUUCUGUUUCAUUG 
                 1127 
                 1593-1615 
               
               
                   
               
               
                 AD-1231061 
                 UGAAACAGAAAUAAACUUCCA 
                 1006 
                 1596-1616 
                 UGGAAGUUUAUUUCUGUUUCAUU 
                 1128 
                 1594-1616 
               
               
                   
               
               
                 AD-1231062 
                 CUCAAACAAGCACUCACGAUA 
                 1007 
                 1618-1638 
                 UAUCGUGAGUGCUUGUUUGAGCA 
                 1129 
                 1616-1638 
               
               
                   
               
               
                 AD-1231063 
                 AAACAAGCACUCACGAUUGUA 
                 1008 
                 1621-1641 
                 UACAAUCGUGAGUGCUUGUUUGA 
                 1130 
                 1619-1641 
               
               
                   
               
               
                 AD-1231064 
                 AACAAGCACUCACGAUUGUUA 
                 1009 
                 1622-1642 
                 UAACAAUCGUGAGUGCUUGUUUG 
                 1131 
                 1620-1642 
               
               
                   
               
               
                 AD-1231074 
                 CUAGCAUUGGAAAAUGUUGUA 
                 1010 
                 2002-2022 
                 UACAACAUUUUCCAAUGCUAGGG 
                 1132 
                 2000-2022 
               
               
                   
               
               
                 AD-1231078 
                 CCAGAACAAGAAUUCUUUUGA 
                 1011 
                 2091-2111 
                 UCAAAAGAAUUCUUGUUCUGGUC 
                 1133 
                 2089-2111 
               
               
                   
               
               
                 AD-1231079 
                 CAGAACAAGAAUUCUUUUGUA 
                 1012 
                 2092-2112 
                 UACAAAAGAAUUCUUGUUCUGGU 
                 1134 
                 2090-2112 
               
               
                   
               
               
                 AD-1231084 
                 CUAGGGAAAGUCAUUCAGUGA 
                 1013 
                 255-275 
                 UCACUGAAUGACUUUCCCUAGAC 
                 1135 
                 253-275 
               
               
                   
               
               
                 AD-1231086 
                 UGCACCUAAAAAUGUGUCUGA 
                 1014 
                 2358-2378 
                 UCAGACACAUUUUUAGGUGCAGU 
                 1136 
                 2356-2378 
               
               
                   
               
               
                 AD-1231088 
                 GUAUCAAUGAUGCUUUCCGUA 
                 1015 
                 2429-2449 
                 UACGGAAAGCAUCAUUGAUACGG 
                 1137 
                 2427-2449 
               
               
                   
               
               
                 AD-1231089 
                 AGAAAAUAAUCCAGGAUUCCA 
                 1016 
                 2667-2687 
                 UGGAAUCCUGGAUUAUUUUCUCC 
                 1138 
                 2665-2687 
               
               
                   
               
               
                 AD-1231090 
                 AAUCCAGGAUUCCAAAACACA 
                 1017 
                 2674-2694 
                 UGUGUUUUGGAAUCCUGGAUUAU 
                 1139 
                 2672-2694 
               
               
                   
               
               
                 AD-1231094 
                 UCCUUUUAGAAAAAUCUAUGA 
                 1018 
                 2710-2730 
                 UCAUAGAUUUUUCUAAAAGGAGG 
                 1140 
                 2708-2730 
               
               
                   
               
               
                 AD-1231095 
                 CCUCUUGAGGUGAUUUUGUUA 
                 1019 
                 2735-2755 
                 UAACAAAAUCACCUCAAGAGGAA 
                 1141 
                 2733-2755 
               
               
                   
               
               
                 AD-1231096 
                 AGCAUCUUCAUUGACAUUGCA 
                 1020 
                 2873-2893 
                 UGCAAUGUCAAUGAAGAUGCUCU 
                 1142 
                 2871-2893 
               
               
                   
               
               
                 AD-1231097 
                 GCAUCUUCAUUGACAUUGCUA 
                 1021 
                 2874-2894 
                 UAGCAAUGUCAAUGAAGAUGCUC 
                 1143 
                 2872-2894 
               
               
                   
               
               
                 AD-1231105 
                 UUAUUUCUGUCUCUGGAUUUA 
                 1022 
                 2902-2922 
                 UAAAUCCAGAGACAGAAAUAAAU 
                 1144 
                 2900-2922 
               
               
                   
               
               
                 AD-1231106 
                 UUUCUGUCUCUGGAUUUGACA 
                 1023 
                 2905-2925 
                 UGUCAAAUCCAGAGACAGAAAUA 
                 1145 
                 2903-2925 
               
               
                   
               
               
                 AD-1231107 
                 UCUCUGGAUUUGACUUCUGUA 
                 1024 
                 2911-2931 
                 UACAGAAGUCAAAUCCAGAGACA 
                 1146 
                 2909-2931 
               
               
                   
               
               
                 AD-1231109 
                 UGUUCAGGGAUAAUCUAAAUA 
                 1025 
                 2997-3017 
                 UAUUUAGAUUAUCCCUGAACAGC 
                 1147 
                 2995-3017 
               
               
                   
               
               
                 AD-1231110 
                 AAAUGUAAAUGUCUGUUGAAA 
                 1026 
                 3013-3033 
                 UUUCAACAGACAUUUACAUUUAG 
                 1148 
                 3011-3033 
               
               
                   
               
               
                 AD-1231111 
                 AAUGUCUGUUGAAUUUCUGAA 
                 1027 
                 3020-3040 
                 UUCAGAAAUUCAACAGACAUUUA 
                 1149 
                 3018-3040 
               
               
                   
               
               
                 AD-1231112 
                 UCAAGUACUAUGGUGAUUUGA 
                 1028 
                 3222-3242 
                 UCAAAUCACCAUAGUACUUGAAC 
                 1150 
                 3220-3242 
               
               
                   
               
               
                 AD-1231113 
                 CAAGUACUAUGGUGAUUUGCA 
                 1029 
                 3223-3243 
                 UGCAAAUCACCAUAGUACUUGAA 
                 1151 
                 3221-3243 
               
               
                   
               
               
                 AD-1231114 
                 UUCAAGGUGACAGGUCUAAAA 
                 1030 
                 3275-3295 
                 UUUUAGACCUGUCACCUUGAAGA 
                 1152 
                 3273-3295 
               
               
                   
               
               
                 AD-1231115 
                 GAAAUUCAGAAUCUCACAGUA 
                 1031 
                 559-579 
                 UACUGUGAGAUUCUGAAUUUCUU 
                 1153 
                 557-579 
               
               
                   
               
               
                 AD-1231116 
                 GAGGACAUUGCUUUUUCACUA 
                 1032 
                 3320-3340 
                 UAGUGAAAAAGCAAUGUCCUCUA 
                 1154 
                 3318-3340 
               
               
                   
               
               
                 AD-1231117 
                 GGACAUUGCUUUUUCACUUCA 
                 1033 
                 3322-3342 
                 UGAAGUGAAAAAGCAAUGUCCUC 
                 1155 
                 3320-3342 
               
               
                   
               
               
                 AD-1231119 
                 GAGUGAAUGGAAAUUCCAACA 
                 1034 
                 3452-3472 
                 UGUUGGAAUUUCCAUUCACUCCA 
                 1156 
                 3450-3472 
               
               
                   
               
               
                 AD-1231120 
                 UGAAUGGAAAUUCCAACUGUA 
                 1035 
                 3455-3475 
                 UACAGUUGGAAUUUCCAUUCACU 
                 1157 
                 3453-3475 
               
               
                   
               
            
           
         
       
     
     
       
         
           
               
             
               
                 TABLE 5 
               
             
            
               
                   
               
               
                 Modified Sense and Antisense Strand Cynomolgus Monkey ACE2 dsRNA Sequences 
               
            
           
           
               
               
               
               
               
               
               
            
               
                   
                   
                 SEQ 
                   
                 SEQ 
                   
                 SEQ 
               
               
                   
                 Sense 
                 ID 
                 Antisense 
                 ID 
                   
                 ID 
               
               
                 Duplex Name 
                 Sequence 5′ to 3′ 
                 NO: 
                 Sequence 5′ to 3′ 
                 NO: 
                 mRNA target sequence 
                 NO: 
               
               
                   
               
               
                 AD-1230824 
                 csusaag(Chd)Afu 
                 1158 
                 VPusAfsaugGfauu 
                 1277 
                 ACCUAAGCAUUUAAA 
                 1396 
               
               
                   
                 UfUfAfaaauccaus 
                   
                 uuaaAfuGfcuuags 
                   
                 AUCCAUUG 
                   
               
               
                   
                 usa 
                   
                 gsu 
                   
                   
                   
               
               
                   
               
               
                 AD-1230830 
                 csasucu(Uhd)Cfa 
                 1159 
                 VPusAfsagcAfaug 
                 1278 
                 AGCAUCUUCAUUGAC 
                 1397 
               
               
                   
                 UfUfGfacauugcus 
                   
                 ucaaUfgAfagaugs 
                   
                 AUUGCUUU 
                   
               
               
                   
                 usa 
                   
                 csu 
                   
                   
                   
               
               
                   
               
               
                 AD-1230834 
                 csusgggaAfaAfUf 
                 1160 
                 VPusUfsuagCfaug 
                 1279 
                 UUCUGGGAAAAUUCC 
                 1398 
               
               
                   
                 Ufcca(Uhd)gcuas 
                   
                 gaauUfuUfcccags 
                   
                 AUGCUAAC 
                   
               
               
                   
                 asa 
                   
                 asa 
                   
                   
                   
               
               
                   
               
               
                 AD-1230835 
                 cscsuaag(Chd)aU 
                 1161 
                 VPusAfsuggAfuuu 
                 1280 
                 CACCUAAGCAUUUAA 
                 1399 
               
               
                   
                 fUfUfaaaauccasu 
                   
                 uaaaUfgCfuuaggs 
                   
                 AAUCCAUU 
                   
               
               
                   
                 sa 
                   
                 usg 
                   
                   
                   
               
               
                   
               
               
                 AD-1230840 
                 gsasaga(Chd)Cfu 
                 1162 
                 VPusCfsuuuGfaua 
                 1281 
                 CCGAAGACCUGUUCU 
                 1400 
               
               
                   
                 GfUfUfcuaucaaas 
                   
                 gaacAfgGfucuucs 
                   
                 AUCAAAGU 
                   
               
               
                   
                 gsa 
                   
                 gsg 
                   
                   
                   
               
               
                   
               
               
                 AD-1230850 
                 asasaug(Uhd)Gfa 
                 407 
                 VPusGfsaguUfuga 
                 584 
                 ACAAAUGUGACAUCU 
                 1401 
               
               
                   
                 CfAfUfcucaaacus 
                   
                 gaugUfcAfcauuus 
                   
                 CAAACUCU 
                   
               
               
                   
                 csa 
                   
                 gsu 
                   
                   
                   
               
               
                   
               
               
                 AD-1230860 
                 usasaaug(Uhd)cU 
                 1163 
                 VPusAfsgaaAfuuc 
                 1282 
                 UGUAAAUGUCUGUUG 
                 1402 
               
               
                   
                 fGfUfugaauuucsu 
                   
                 aacaGfaCfauuuas 
                   
                 AAUUUCUG 
                   
               
               
                   
                 sa 
                   
                 csa 
                   
                   
                   
               
               
                   
               
               
                 AD-1230861 
                 gscsuca(Chd)Ufu 
                 1164 
                 VPusUfsggaUfuaa 
                 1283 
                 UAGUUCACUUUCAUU 
                 1403 
               
               
                   
                 UfCfAfuuuaauccs 
                   
                 augaAfaGfugagcs 
                   
                 UAAUCCAU 
                   
               
               
                   
                 asa 
                   
                 usa 
                   
                   
                   
               
               
                   
               
               
                 AD-1230862 
                 csascuu(Uhd)Cfa 
                 1165 
                 VPusCfsaauGfgau 
                 1284 
                 UUCACUUUCAUUUAA 
                 1404 
               
               
                   
                 UfUfUfaauccauus 
                   
                 uaaaUfgAfaagugs 
                   
                 UCCAUUGU 
                   
               
               
                   
                 gsa 
                   
                 asg 
                   
                   
                   
               
               
                   
               
               
                 AD-1230863 
                 asusugc(Uhd)Ufu 
                 1166 
                 VPusCfsuugGfaag 
                 1285 
                 ACAUUGCUUUUUCAC 
                 1405 
               
               
                   
                 UfUfCfacuuccaas 
                   
                 ugaaAfaAfgcaaus 
                   
                 UUCCAAGC 
                   
               
               
                   
                 gsa 
                   
                 gsu 
                   
                   
                   
               
               
                   
               
               
                 AD-1230864 
                 csasgaa(Uhd)Cfu 
                 1167 
                 VPusAfsgcuUfgac 
                 1286 
                 UUCAGAAUCUCACAG 
                 1406 
               
               
                   
                 CfAfCfagucaagcs 
                   
                 ugugAfgAfuucugs 
                   
                 UCAAGCUU 
                   
               
               
                   
                 usa 
                   
                 asa 
                   
                   
                   
               
               
                   
               
               
                 AD-1230865 
                 asgsaau(Chd)Ufc 
                 1168 
                 VPusAfsagcUfuga 
                 1287 
                 UCAGAAUCUCACAGU 
                 1407 
               
               
                   
                 AfCfAfgucaagcus 
                   
                 cuguGfaGfauucus 
                   
                 CAAGCUUC 
                   
               
               
                   
                 usa 
                   
                 gsa 
                   
                   
                   
               
               
                   
               
               
                 AD-1230866 
                 csasguc(Uhd)Cfu 
                 1169 
                 VPusAfscaaAfaga 
                 1288 
                 UCCAGUCUCUUAAAU 
                 1408 
               
               
                   
                 UfAfAfaucuuuugs 
                   
                 uuuaAfgAfgacugs 
                   
                 CUUUUGUA 
                   
               
               
                   
                 usa 
                   
                 gsg 
                   
                   
                   
               
               
                   
               
               
                 AD-1230871 
                 csusucc(Uhd)Afa 
                 420 
                 VPusCfscuuGfagu 
                 597 
                 GUCUUCCUAAUAUGA 
                 1409 
               
               
                   
                 UfAfUfgacucaags 
                   
                 cauaUfuAfggaags 
                   
                 CUCAAGGA 
                   
               
               
                   
                 gsa 
                   
                 asc 
                   
                   
                   
               
               
                   
               
               
                 AD-1230872 
                 usgsggaaAfaUfUf 
                 1170 
                 VPusGfsuuaGfcau 
                 1289 
                 UCUGGGAAAAUUCCA 
                 1410 
               
               
                   
                 Cfcaug(Chd)uaas 
                   
                 ggaaUfuUfucccas 
                   
                 UGCUAACU 
                   
               
               
                   
                 csa 
                   
                 gsa 
                   
                   
                   
               
               
                   
               
               
                 AD-1230893 
                 csuscaaa(Chd)uC 
                 1171 
                 VPusCfsagcUfucu 
                 1290 
                 AUCUCAAACUCUACA 
                 1411 
               
               
                   
                 fUfAfcagaagcusg 
                   
                 guagAfgUfuugags 
                   
                 GAAGCUGG 
                   
               
               
                   
                 sa 
                   
                 asu 
                   
                   
                   
               
               
                   
               
               
                 AD-1230898 
                 cscsaagaAfuCfUf 
                 442 
                 VPusAfsaauUfaaa 
                 619 
                 AACCAAGAAUCUCCU 
                 1412 
               
               
                   
                 Cfcuu(Uhd)aauus 
                   
                 ggagAfuUfcuuggs 
                   
                 UUAAUUUC 
                   
               
               
                   
                 usa 
                   
                 usu 
                   
                   
                   
               
               
                   
               
               
                 AD-1230899 
                 csasagaa(Uhd)cU 
                 1172 
                 VPusGfsaaaUfuaa 
                 1291 
                 ACCAAGAAUCUCCUU 
                 1413 
               
               
                   
                 fCfCfuuuaauuusc 
                   
                 aggaGfaUfucuugs 
                   
                 UAAUUUCU 
                   
               
               
                   
                 sa 
                   
                 gsu 
                   
                   
                   
               
               
                   
               
               
                 AD-1230900 
                 asasgaa(Uhd)Cfu 
                 1173 
                 VPusAfsgaaAfuua 
                 1292 
                 CCAAGAAUCUCCUUU 
                 1414 
               
               
                   
                 CfCfUfuuaauuucs 
                   
                 aaggAfgAfuucuus 
                   
                 AAUUUCUA 
                   
               
               
                   
                 usa 
                   
                 gsg 
                   
                   
                   
               
               
                   
               
               
                 AD-1230901 
                 csusgca(Chd)Cfu 
                 1174 
                 VPusAfsgacAfcau 
                 1293 
                 CACUGCACCUAAAAA 
                 1415 
               
               
                   
                 AfAfAfaaugugucs 
                   
                 uuuuAfgGfugcags 
                   
                 UGUGUCUG 
                   
               
               
                   
                 usa 
                   
                 usg 
                   
                   
                   
               
               
                   
               
               
                 AD-1230903 
                 uscsacu(Uhd)Ufc 
                 1175 
                 VPusAfsaugGfauu 
                 1294 
                 GUUCACUUUCAUUUA 
                 1416 
               
               
                   
                 AfUfUfuaauccaus 
                   
                 aaauGfaAfagugas 
                   
                 AUCCAUUG 
                   
               
               
                   
                 usa 
                   
                 gsc 
                   
                   
                   
               
               
                   
               
               
                 AD-1230905 
                 asasuu(Chd)aGfa 
                 1176 
                 VPusUfsgacUfgug 
                 1295 
                 GAAAUUCAGAAUCUC 
                 1417 
               
               
                   
                 AfUfCfucacagucs 
                   
                 agauUfcUfgaauus 
                   
                 ACAGUCAA 
                   
               
               
                   
                 asa 
                   
                 usc 
                   
                   
                   
               
               
                   
               
               
                 AD-1230906 
                 asusucagAfaUfCf 
                 1177 
                 VPusUfsugaCfugu 
                 1296 
                 AAAUUCAGAAUCUCA 
                 1418 
               
               
                   
                 Ufcacag(Uhd)cas 
                   
                 gagaUfuCfugaaus 
                   
                 CAGUCAAG 
                   
               
               
                   
                 asa 
                   
                 usu 
                   
                   
                   
               
               
                   
               
               
                 AD-1230907 
                 ususcagaAfuCfUf 
                 1178 
                 VPusCfsuugAfcug 
                 1297 
                 AAUUCAGAAUCUCAC 
                 1419 
               
               
                   
                 Cfacag(Uhd)caas 
                   
                 ugagAfuUfcugaas 
                   
                 AGUCAAGC 
                   
               
               
                   
                 gsa 
                   
                 usu 
                   
                   
                   
               
               
                   
               
               
                 AD-1230908 
                 gscscuga(Uhd)aG 
                 1179 
                 VPusGfsaaaUfgag 
                 1298 
                 AUGCCUGAUAGAAAC 
                 1420 
               
               
                   
                 fAfAfacucauuusc 
                   
                 uuucUfaUfcaggcs 
                   
                 UCAUUUCC 
                   
               
               
                   
                 sa 
                   
                 asu 
                   
                   
                   
               
               
                   
               
               
                 AD-1230909 
                 cscsagg(Uhd)Ufu 
                 1180 
                 VPusAfsuuaUfuuc 
                 1299 
                 AUCCAGGUUUGAAUG 
                 1421 
               
               
                   
                 GfAfAfugaaauaas 
                   
                 auucAfaAfccuggs 
                   
                 AAAUAAUG 
                   
               
               
                   
                 usa 
                   
                 usu 
                   
                   
                   
               
               
                   
               
               
                 AD-1230917 
                 uscsugggAfaAfAf 
                 1181 
                 VPusUfsagcAfugg 
                 1300 
                 AUUCUGGGAAAAUUC 
                 1422 
               
               
                   
                 Ufucca(Uhd)gcus 
                   
                 aauuUfuCfccagas 
                   
                 CAUGCUAA 
                   
               
               
                   
                 asa 
                   
                 asu 
                   
                   
                   
               
               
                   
               
               
                 AD-1230918 
                 csascaa(Chd)Cfu 
                 1182 
                 VPusUfscuuAfgca 
                 1301 
                 UGCACAACCUUUUCU 
                 1423 
               
               
                   
                 UfUfUfcugcuaags 
                   
                 gaaaAfgGfuugugs 
                   
                 GCUAAGAA 
                   
               
               
                   
                 asa 
                   
                 csa 
                   
                   
                   
               
               
                   
               
               
                 AD-1230919 
                 csasacc(Uhd)Ufu 
                 1183 
                 VPusUfsuucUfuag 
                 1302 
                 CACAACCUUUUCUGC 
                 1424 
               
               
                   
                 UfCfUfgcuaagaas 
                   
                 cagaAfaAfgguugs 
                   
                 UAAGAAAU 
                   
               
               
                   
                 asa 
                   
                 usg 
                   
                   
                   
               
               
                   
               
               
                 AD-1230924 
                 gsasaa(Chd)aGfa 
                 1184 
                 VPusAfsggaAfguu 
                 1303 
                 AUGAAACAGAAAUAA 
                 1425 
               
               
                   
                 AfAfUfaaacuuccs 
                   
                 uauuUfcUfguuucs 
                   
                 ACUUCCUG 
                   
               
               
                   
                 usa 
                   
                 asu 
                   
                   
                   
               
               
                   
               
               
                 AD-1230925 
                 uscsaaa(Chd)Afa 
                 1185 
                 VPusAfsaucGfuga 
                 1304 
                 GCUCAAACAAGCACU 
                 1426 
               
               
                   
                 GfCfAfcucacgaus 
                   
                 gugcUfuGfuuugas 
                   
                 CACGAUUG 
                   
               
               
                   
                 usa 
                   
                 gsc 
                   
                   
                   
               
               
                   
               
               
                 AD-1230926 
                 uscsugc(Chd)Afu 
                 1186 
                 VPusAfscauGfuaa 
                 1305 
                 ACUCUGCCAUUUACU 
                 1427 
               
               
                   
                 UfUfAfcuuacaugs 
                   
                 guaaAfuGfgcagas 
                   
                 UACAUGUU 
                   
               
               
                   
                 usa 
                   
                 gsu 
                   
                   
                   
               
               
                   
               
               
                 AD-1230927 
                 csusgcca(Uhd)uU 
                 1187 
                 VPusAfsacaUfgua 
                 1306 
                 CUCUGCCAUUUACUU 
                 1428 
               
               
                   
                 fAfCfuuacaugusu 
                   
                 aguaAfaUfggcags 
                   
                 ACAUGUUA 
                   
               
               
                   
                 sa 
                   
                 asg 
                   
                   
                   
               
               
                   
               
               
                 AD-1230929 
                 csgsaaga(Chd)cU 
                 1188 
                 VPusUfsuugAfuag 
                 1307 
                 GCCGAAGACCUGUUC 
                 1429 
               
               
                   
                 fGfUfucuaucaasa 
                   
                 aacaGfgUfcuucgs 
                   
                 UAUCAAAG 
                   
               
               
                   
                 sa 
                   
                 gsc 
                   
                   
                   
               
               
                   
               
               
                 AD-1230933 
                 asasgac(Chd)Ufg 
                 1189 
                 VPusAfscuuUfgau 
                 1308 
                 CGAAGACCUGUUCUA 
                 1430 
               
               
                   
                 UfUfCfuaucaaags 
                   
                 agaaCfaGfgucuus 
                   
                 UCAAAGUU 
                   
               
               
                   
                 usa 
                   
                 csg 
                   
                   
                   
               
               
                   
               
               
                 AD-1230943 
                 asuscaa(Uhd)Gfa 
                 1190 
                 VPusAfsgacGfgaa 
                 1309 
                 GUAUCAAUGAUGCUU 
                 1431 
               
               
                   
                 UfGfCfuuuccgucs 
                   
                 agcaUfcAfuugaus 
                   
                 UCCGUCUG 
                   
               
               
                   
                 usa 
                   
                 asc 
                   
                   
                   
               
               
                   
               
               
                 AD-1230944 
                 asasugu(Chd)Cfa 
                 1191 
                 VPusUfsuauUfcau 
                 1310 
                 AGAAUGUCCAAAACA 
                 1432 
               
               
                   
                 AfAfAfcaugaauas 
                   
                 guuuUfgGfacauus 
                   
                 UGAAUAAU 
                   
               
               
                   
                 asa 
                   
                 csu 
                   
                   
                   
               
               
                   
               
               
                 AD-1230945 
                 asusguc(Chd)Afa 
                 1192 
                 VPusAfsuuaUfuca 
                 1311 
                 GAAUGUCCAAAACAU 
                 1433 
               
               
                   
                 AfAfCfaugaauaas 
                   
                 uguuUfuGfgacaus 
                   
                 GAAUAAUG 
                   
               
               
                   
                 usa 
                   
                 usc 
                   
                   
                   
               
               
                   
               
               
                 AD-1230952 
                 uscsugu(Chd)Ufc 
                 1193 
                 VPusAfsaguCfaaa 
                 1312 
                 UUUCUGUCUCUGGAU 
                 1434 
               
               
                   
                 UfGfGfauuugacus 
                   
                 uccaGfaGfacagas 
                   
                 UUGACUUC 
                   
               
               
                   
                 usa 
                   
                 asa 
                   
                   
                   
               
               
                   
               
               
                 AD-1230953 
                 uscsugga(Uhd)uU 
                 1194 
                 VPusGfsaacAfgaa 
                 1313 
                 UCUCUGGAUUUGACU 
                 1435 
               
               
                   
                 fGfAfcuucuguusc 
                   
                 gucaAfaUfccagas 
                   
                 UCUGUUCU 
                   
               
               
                   
                 sa 
                   
                 gsa 
                   
                   
                   
               
               
                   
               
               
                 AD-1230954 
                 gsasuu(Uhd)gAfc 
                 1195 
                 VPusAfsacaGfaac 
                 1314 
                 UGGAUUUGACUUCUG 
                 1436 
               
               
                   
                 UfUfCfuguucugus 
                   
                 agaaGfuCfaaaucs 
                   
                 UUCUGUUU 
                   
               
               
                   
                 usa 
                   
                 csa 
                   
                   
                   
               
               
                   
               
               
                 AD-1230955 
                 csasggga(Uhd)aA 
                 1196 
                 VPusUfsuacAfuuu 
                 1315 
                 UUCAGGGAUAAUCUA 
                 1437 
               
               
                   
                 fUfCfuaaauguasa 
                   
                 agauUfaUfcccugs 
                   
                 AAUGUAAA 
                   
               
               
                   
                 sa 
                   
                 asa 
                   
                   
                   
               
               
                   
               
               
                 AD-1230956 
                 asgsgga(Uhd)Afa 
                 1197 
                 VPusUfsuuaCfauu 
                 1316 
                 UCAGGGAUAAUCUAA 
                 1438 
               
               
                   
                 UfCfUfaaauguaas 
                   
                 uagaUfuAfucccus 
                   
                 AUGUAAAU 
                   
               
               
                   
                 asa 
                   
                 gsa 
                   
                   
                   
               
               
                   
               
               
                 AD-1230957 
                 usasaaug(Uhd)aA 
                 1198 
                 VPusUfscaaCfaga 
                 1317 
                 UCUAAAUGUAAAUGU 
                 1439 
               
               
                   
                 fAfUfgucuguugsa 
                   
                 cauuUfaCfauuuas 
                   
                 CUGUUGAA 
                   
               
               
                   
                 sa 
                   
                 gsa 
                   
                   
                   
               
               
                   
               
               
                 AD-1230958 
                 asusguc(Uhd)Gfu 
                 1199 
                 VPusUfsucaGfaaa 
                 1318 
                 AAAUGUCUGUUGAAU 
                 1440 
               
               
                   
                 UfGfAfauuucugas 
                   
                 uucaAfcAfgacaus 
                   
                 UUCUGAAG 
                   
               
               
                   
                 asa 
                   
                 usu 
                   
                   
                   
               
               
                   
               
               
                 AD-1230959 
                 uscsugu(Uhd)Gfa 
                 1200 
                 VPusAfsacuUfcag 
                 1319 
                 UGUCUGUUGAAUUUC 
                 1441 
               
               
                   
                 AfUfUfucugaagus 
                   
                 aaauUfcAfacagas 
                   
                 UGAAGUUG 
                   
               
               
                   
                 usa 
                   
                 csa 
                   
                   
                   
               
               
                   
               
               
                 AD-1230960 
                 csuscac(Uhd)Ufu 
                 1201 
                 VPusAfsuggAfuua 
                 1320 
                 AGUUCACUUUCAUUU 
                 1442 
               
               
                   
                 CfAfUfuuaauccas 
                   
                 aaugAfaAfgugags 
                   
                 AAUCCAUU 
                   
               
               
                   
                 usa 
                   
                 csu 
                   
                   
                   
               
               
                   
               
               
                 AD-1230961 
                 ascsuuu(Chd)Afu 
                 1202 
                 VPusAfscaaUfgga 
                 1321 
                 UCACUUUCAUUUAAU 
                 1443 
               
               
                   
                 UfUfAfauccauugs 
                   
                 uuaaAfuGfaaagus 
                   
                 CCAUUGUU 
                   
               
               
                   
                 usa 
                   
                 gsa 
                   
                   
                   
               
               
                   
               
               
                 AD-1230963 
                 gsasauc(Uhd)Cfa 
                 1203 
                 VPusGfsaagCfuug 
                 1322 
                 CAGAAUCUCACAGUC 
                 1444 
               
               
                   
                 CfAfGfucaagcuus 
                   
                 acugUfgAfgauucs 
                   
                 AAGCUUCA 
                   
               
               
                   
                 csa 
                   
                 usg 
                   
                   
                   
               
               
                   
               
               
                 AD-1230964 
                 asasuuc(Chd)Afa 
                 1204 
                 VPusGfsugaAfcau 
                 1323 
                 GAAAUUCCAACUGUA 
                 1445 
               
               
                   
                 CfUfGfuauguucas 
                   
                 acagUfuGfgaauus 
                   
                 UGUUCACC 
                   
               
               
                   
                 csa 
                   
                 usc 
                   
                   
                   
               
               
                   
               
               
                 AD-1230965 
                 asgsucu(Chd)Ufu 
                 1205 
                 VPusUfsacaAfaag 
                 1324 
                 CCAGUCUCUUAAAUC 
                 1446 
               
               
                   
                 AfAfAfucuuuugus 
                   
                 auuuAfaGfagacus 
                   
                 UUUUGUAU 
                   
               
               
                   
                 asa 
                   
                 gsg 
                   
                   
                   
               
               
                   
               
               
                 AD-1230966 
                 gsuscuc(Uhd)Ufa 
                 1206 
                 VPusAfsuacAfaaa 
                 1325 
                 CAGUCUCUUAAAUCU 
                 1447 
               
               
                   
                 AfAfUfcuuuuguas 
                   
                 gauuUfaAfgagacs 
                   
                 UUUGUAUU 
                   
               
               
                   
                 usa 
                   
                 usg 
                   
                   
                   
               
               
                   
               
               
                 AD-1230967 
                 csascu(Chd)aAfu 
                 1207 
                 VPusAfsaucUfagc 
                 1326 
                 GACACUCAAUAAAUG 
                 1448 
               
               
                   
                 AfAfAfugcuagaus 
                   
                 auuuAfuUfgagugs 
                   
                 CUAGAUUU 
                   
               
               
                   
                 usa 
                   
                 usc 
                   
                   
                   
               
               
                   
               
               
                 AD-1230968 
                 ascsucaa(Uhd)aA 
                 1208 
                 VPusAfsaauCfuag 
                 1327 
                 ACACUCAAUAAAUGC 
                 1449 
               
               
                   
                 fAfUfgcuagauusu 
                   
                 cauuUfaUfugagus 
                   
                 UAGAUUUG 
                   
               
               
                   
                 sa 
                   
                 gsu 
                   
                   
                   
               
               
                   
               
               
                 AD-1230974 
                 ascscagg(Uhd)uU 
                 1209 
                 VPusUfsuauUfuca 
                 1328 
                 GAUCCAGGUUUGAAU 
                 1450 
               
               
                   
                 fGfAfaugaaauasa 
                   
                 uucaAfaCfcuggus 
                   
                 GAAAUAAU 
                   
               
               
                   
                 sa 
                   
                 usc 
                   
                   
                   
               
               
                   
               
               
                 AD-1230975 
                 ascscau(Uhd)Afu 
                 1210 
                 VPusGfsaagAfugu 
                 1329 
                 AAACCAUUAUAUGAA 
                 1451 
               
               
                   
                 AfUfGfaacaucuus 
                   
                 ucauAfuAfauggus 
                   
                 CAUCUUCA 
                   
               
               
                   
                 csa 
                   
                 usu 
                   
                   
                   
               
               
                   
               
               
                 AD-1230976 
                 escsauua(Uhd)aU 
                 1211 
                 VPusUfsgaaGfaug 
                 1330 
                 AACCAUUAUAUGAAC 
                 1452 
               
               
                   
                 fGfAfacaucuucsa 
                   
                 uucaUfaUfaauggs 
                   
                 AUCUUCAU 
                   
               
               
                   
                 sa 
                   
                 usu 
                   
                   
                   
               
               
                   
               
               
                 AD-1230985 
                 uscsuaggGfaAfAf 
                 1212 
                 VPusAfscugAfaug 
                 1331 
                 AGUCUAGGGAAAGUC 
                 1453 
               
               
                   
                 Gfucau(Uhd)cags 
                   
                 acuuUfcCfcuagas 
                   
                 AUUCAGUG 
                   
               
               
                   
                 usa 
                   
                 csu 
                   
                   
                   
               
               
                   
               
               
                 AD-1230991 
                 gscsacaa(Chd)cU 
                 1213 
                 VPusCfsuuaGfcag 
                 1332 
                 CUGCACAACCUUUUC 
                 1454 
               
               
                   
                 fUfUfucugcuaasg 
                   
                 aaaaGfgUfugugcs 
                   
                 UGCUAAGA 
                   
               
               
                   
                 sa 
                   
                 asg 
                   
                   
                   
               
               
                   
               
               
                 AD-1230995 
                 usgscca(Uhd)Ufu 
                 1214 
                 VPusUfsaacAfugu 
                 1333 
                 UCUGCCAUUUACUUA 
                 1455 
               
               
                   
                 AfCfUfuacauguus 
                   
                 aaguAfaAfuggcas 
                   
                 CAUGUUAG 
                   
               
               
                   
                 asa 
                   
                 gsa 
                   
                   
                   
               
               
                   
               
               
                 AD-1231000 
                 gsasccagAfaCfAf 
                 1215 
                 VPusAfsaagAfauu 
                 1334 
                 AAGACCAGAACAAGA 
                 1456 
               
               
                   
                 Afgaau(Uhd)cuus 
                   
                 cuugUfuCfuggucs 
                   
                 AUUCUUUU 
                   
               
               
                   
                 usa 
                   
                 usu 
                   
                   
                   
               
               
                   
               
               
                 AD-1231005 
                 usasucaa(Uhd)gA 
                 1216 
                 VPusGfsacgGfaaa 
                 1335 
                 CGUAUCAAUGAUGCU 
                 1457 
               
               
                   
                 fUfGfcuuuccgusc 
                   
                 gcauCfaUfugauas 
                   
                 UUCCGUCU 
                   
               
               
                   
                 sa 
                   
                 csg 
                   
                   
                   
               
               
                   
               
               
                 AD-1231006 
                 usgsuc(Chd)aAfa 
                 1217 
                 VPusCfsauuAfuuc 
                 1336 
                 AAUGUCCAAAACAUG 
                 1458 
               
               
                   
                 AfCfAfugaauaaus 
                   
                 auguUfuUfggacas 
                   
                 AAUAAUGC 
                   
               
               
                   
                 gsa 
                   
                 usu 
                   
                   
                   
               
               
                   
               
               
                 AD-1231007 
                 csusccu(Uhd)Ufu 
                 1218 
                 VPusAfsuagAfuuu 
                 1337 
                 ACCUCCUUUUAGAAA 
                 1459 
               
               
                   
                 AfGfAfaaaaucuas 
                   
                 uucuAfaAfaggags 
                   
                 AAUCUAUG 
                   
               
               
                   
                 usa 
                   
                 gsu 
                   
                   
                   
               
               
                   
               
               
                 AD-1231008 
                 asusugu(Chd)Cfa 
                 1219 
                 VPusCfscauGfuug 
                 1338 
                 AAAUUGUCCAAAGAC 
                 1460 
               
               
                   
                 AfAfGfacaacaugs 
                   
                 ucuuUfgGfacaaus 
                   
                 AACAUGGU 
                   
               
               
                   
                 gsa 
                   
                 usu 
                   
                   
                   
               
               
                   
               
               
                 AD-1231013 
                 ususcug(Uhd)Cfu 
                 1220 
                 VPusAfsgucAfaau 
                 1339 
                 AUUUCUGUCUCUGGA 
                 1461 
               
               
                   
                 CfUfGfgauuugacs 
                   
                 ccagAfgAfcagaas 
                   
                 UUUGACUU 
                   
               
               
                   
                 usa 
                   
                 asu 
                   
                   
                   
               
               
                   
               
               
                 AD-1231014 
                 csuscuggAfuUfUf 
                 1221 
                 VPusAfsacaGfaag 
                 1340 
                 GUCUCUGGAUUUGAC 
                 1462 
               
               
                   
                 Gfacuu(Chd)ugus 
                   
                 ucaaAfuCfcagags 
                   
                 UUCUGUUC 
                   
               
               
                   
                 usa 
                   
                 asc 
                   
                   
                   
               
               
                   
               
               
                 AD-1231015 
                 csusgga(Uhd)Ufu 
                 1222 
                 VPusAfsgaaCfaga 
                 1341 
                 CUCUGGAUUUGACUU 
                 1463 
               
               
                   
                 GfAfCfuucuguucs 
                   
                 agucAfaAfuccags 
                   
                 CUGUUCUG 
                   
               
               
                   
                 usa 
                   
                 asg 
                   
                   
                   
               
               
                   
               
               
                 AD-1231016 
                 usgsgau(Uhd)Ufg 
                 1223 
                 VPusCfsagaAfcag 
                 1342 
                 UCUGGAUUUGACUUC 
                 1464 
               
               
                   
                 AfCfUfucuguucus 
                   
                 aaguCfaAfauccas 
                   
                 UGUUCUGU 
                   
               
               
                   
                 gsa 
                   
                 gsa 
                   
                   
                   
               
               
                   
               
               
                 AD-1231017 
                 gsgsauu(Uhd)Gfa 
                 1224 
                 VPusAfscagAfaca 
                 1343 
                 CUGGAUUUGACUUCU 
                 1465 
               
               
                   
                 CfUfUfcuguucugs 
                   
                 gaagUfcAfaauccs 
                   
                 GUUCUGUU 
                   
               
               
                   
                 usa 
                   
                 asg 
                   
                   
                   
               
               
                   
               
               
                 AD-1231018 
                 gsusaaa(Uhd)Gfu 
                 1225 
                 VPusGfsaaaUfuca 
                 1344 
                 AUGUAAAUGUCUGUU 
                 1466 
               
               
                   
                 CfUfGfuugaauuus 
                   
                 acagAfcAfuuuacs 
                   
                 GAAUUUCU 
                   
               
               
                   
                 csa 
                   
                 asu 
                   
                   
                   
               
               
                   
               
               
                 AD-1231019 
                 asasaug(Uhd)Cfu 
                 1226 
                 VPusCfsagaAfauu 
                 1345 
                 GUAAAUGUCUGUUGA 
                 1467 
               
               
                   
                 GfUfUfgaauuucus 
                   
                 caacAfgAfcauuus 
                   
                 AUUUCUGA 
                   
               
               
                   
                 gsa 
                   
                 asc 
                   
                   
                   
               
               
                   
               
               
                 AD-1231020 
                 usgsucug(Uhd)uG 
                 1227 
                 VPusCfsuucAfgaa 
                 1346 
                 AAUGUCUGUUGAAUU 
                 1468 
               
               
                   
                 fAfAfuuucugaasg 
                   
                 auucAfaCfagacas 
                   
                 UCUGAAGU 
                   
               
               
                   
                 sa 
                   
                 usu 
                   
                   
                   
               
               
                   
               
               
                 AD-1231021 
                 csusgu(Uhd)gAfa 
                 1228 
                 VPusCfsaacUfuca 
                 1347 
                 GUCUGUUGAAUUUCU 
                 1469 
               
               
                   
                 UfUfUfcugaaguus 
                   
                 gaaaUfuCfaacags 
                   
                 GAAGUUGA 
                   
               
               
                   
                 gsa 
                   
                 asc 
                   
                   
                   
               
               
                   
               
               
                 AD-1231023 
                 uscsagaa(Uhd)cU 
                 1229 
                 VPusGfscuuGfacu 
                 1348 
                 AUUCAGAAUCUCACA 
                 1470 
               
               
                   
                 fCfAfcagucaagsc 
                   
                 gugaGfaUfucugas 
                   
                 GUCAAGCU 
                   
               
               
                   
                 sa 
                   
                 asu 
                   
                   
                   
               
               
                   
               
               
                 AD-1231024 
                 asuscuca(Chd)aG 
                 1230 
                 VPusCfsugaAfgcu 
                 1349 
                 GAAUCUCACAGUCAA 
                 1471 
               
               
                   
                 fUfCfaagcuucasg 
                   
                 ugacUfgUfgagaus 
                   
                 GCUUCAGU 
                   
               
               
                   
                 sa 
                   
                 usc 
                   
                   
                   
               
               
                   
               
               
                 AD-1231025 
                 csusacug(Uhd)uC 
                 1231 
                 VPusCfscacAfguu 
                 1350 
                 UUCCACUGUUCUCUA 
                 1472 
               
               
                   
                 fUfCfuaacugugsg 
                   
                 agagAfaCfaguags 
                   
                 ACUGUGGA 
                   
               
               
                   
                 sa 
                   
                 asa 
                   
                   
                   
               
               
                   
               
               
                 AD-1231026 
                 gsasauggAfaAfUf 
                 1232 
                 VPusUfsacaGfuug 
                 1351 
                 GUGAAUGGAAAUUCC 
                 1473 
               
               
                   
                 Ufccaa(Chd)ugus 
                   
                 gaauUfuCfcauucs 
                   
                 AACUGUAU 
                   
               
               
                   
                 asa 
                   
                 asc 
                   
                   
                   
               
               
                   
               
               
                 AD-1231027 
                 gsasaau(Uhd)Cfc 
                 1233 
                 VPusGfsaacAfuac 
                 1352 
                 UGGAAAUUCCAACUG 
                 1474 
               
               
                   
                 AfAfCfuguauguus 
                   
                 aguuGfgAfauuucs 
                   
                 UAUGUUCA 
                   
               
               
                   
                 csa 
                   
                 csa 
                   
                   
                   
               
               
                   
               
               
                 AD-1231028 
                 cscsagu(Chd)Ufc 
                 1234 
                 VPusCfsaaaAfgau 
                 1353 
                 AUCCAGUCUCUUAAA 
                 1475 
               
               
                   
                 UfUfAfaaucuuuus 
                   
                 uuaaGfaGfacuggs 
                   
                 UCUUUUGU 
                   
               
               
                   
                 gsa 
                   
                 gsu 
                   
                   
                   
               
               
                   
               
               
                 AD-1231029 
                 ascsaaag(Chd)aG 
                 1235 
                 VPusUfsuauUfgag 
                 1354 
                 GCACAAAGCAGACAC 
                 1476 
               
               
                   
                 fAfCfacucaauasa 
                   
                 ugucUfgCfuuugus 
                   
                 UCAAUAAA 
                   
               
               
                   
                 sa 
                   
                 gsc 
                   
                   
                   
               
               
                   
               
               
                 AD-1231030 
                 asasag(Chd)aGfa 
                 1236 
                 VPusAfsuuuAfuug 
                 1355 
                 ACAAAGCAGACACUC 
                 1477 
               
               
                   
                 CfAfCfucaauaaas 
                   
                 agugUfcUfgcuuus 
                   
                 AAUAAAUG 
                   
               
               
                   
                 usa 
                   
                 gsu 
                   
                   
                   
               
               
                   
               
               
                 AD-1231031 
                 gscsaga(Chd)Afc 
                 1237 
                 VPusAfsgcaUfuua 
                 1356 
                 AAGCAGACACUCAAU 
                 1478 
               
               
                   
                 UfCfAfauaaaugcs 
                   
                 uugaGfuGfucugcs 
                   
                 AAAUGCUA 
                   
               
               
                   
                 usa 
                   
                 usu 
                   
                   
                   
               
               
                   
               
               
                 AD-1231032 
                 asgsaca(Chd)Ufc 
                 1238 
                 VPusCfsuagCfauu 
                 1357 
                 GCAGACACUCAAUAA 
                 1479 
               
               
                   
                 AfAfUfaaaugcuas 
                   
                 uauuGfaGfugucus 
                   
                 AUGCUAGA 
                   
               
               
                   
                 gsa 
                   
                 gsc 
                   
                   
                   
               
               
                   
               
               
                 AD-1231040 
                 csasuac(Chd)Ufu 
                 1239 
                 VPusUfsuaaUfcuc 
                 1358 
                 AACGUACCUUUGAAG 
                 1480 
               
               
                   
                 UfGfAfagagauuas 
                   
                 uucaAfaGfguaugs 
                   
                 AGAUUAAA 
                   
               
               
                   
                 asa 
                   
                 usu 
                   
                   
                   
               
               
                   
               
               
                 AD-1231041 
                 asusacc(Uhd)Ufu 
                 1240 
                 VPusUfsuuaAfucu 
                 1359 
                 ACGUACCUUUGAAGA 
                 1481 
               
               
                   
                 GfAfAfgagauuaas 
                   
                 cuucAfaAfgguaus 
                   
                 GAUUAAAC 
                   
               
               
                   
                 asa 
                   
                 gsu 
                   
                   
                   
               
               
                   
               
               
                 AD-1231042 
                 ascscuu(Uhd)Gfa 
                 1241 
                 VPusGfsguuUfaau 
                 1360 
                 GUACCUUUGAAGAGA 
                 1482 
               
               
                   
                 AfGfAfgauuaaacs 
                   
                 cucuUfcAfaaggus 
                   
                 UUAAACCA 
                   
               
               
                   
                 csa 
                   
                 asu 
                   
                   
                   
               
               
                   
               
               
                 AD-1231043 
                 csasuua(Uhd)Afu 
                 1242 
                 VPusAfsugaAfgau 
                 1361 
                 ACCAUUAUAUGAACA 
                 1483 
               
               
                   
                 GfAfAfcaucuucas 
                   
                 guucAfuAfuaaugs 
                   
                 UCUUCAUG 
                   
               
               
                   
                 usa 
                   
                 gsu 
                   
                   
                   
               
               
                   
               
               
                 AD-1231056 
                 csascac(Chd)Ufa 
                 1243 
                 VPusAfsuuuUfaaa 
                 1362 
                 GCCACACCUAAGCAU 
                 1484 
               
               
                   
                 AfGfCfauuuaaaas 
                   
                 ugcuUfaGfgugugs 
                   
                 UUAAAAUC 
                   
               
               
                   
                 usa 
                   
                 gsc 
                   
                   
                   
               
               
                   
               
               
                 AD-1231057 
                 ascscuaaGfcAfUf 
                 1244 
                 VPusUfsggaUfuuu 
                 1363 
                 ACACCUAAGCAUUUA 
                 1485 
               
               
                   
                 Ufuaaaa(Uhd)ccs 
                   
                 aaauGfcUfuaggus 
                   
                 AAAUCCAU 
                   
               
               
                   
                 asa 
                   
                 gsu 
                   
                   
                   
               
               
                   
               
               
                 AD-1231059 
                 csasaugaAfaCfAf 
                 1245 
                 VPusAfsguuUfauu 
                 1364 
                 GACAAUGAAACAGAA 
                 1486 
               
               
                   
                 Gfaaa(Uhd)aaacs 
                   
                 ucugUfuUfcauugs 
                   
                 AUAAACUU 
                   
               
               
                   
                 usa 
                   
                 usc 
                   
                   
                   
               
               
                   
               
               
                 AD-1231060 
                 asusgaaa(Chd)aG 
                 1246 
                 VPusGfsaagUfuua 
                 1365 
                 CAAUGAAACAGAAAU 
                 1487 
               
               
                   
                 fAfAfauaaacuusc 
                   
                 uuucUfgUfuucaus 
                   
                 AAACUUCC 
                   
               
               
                   
                 sa 
                   
                 usg 
                   
                   
                   
               
               
                   
               
               
                 AD-1231061 
                 usgsaaa(Chd)Afg 
                 1247 
                 VPusGfsgaaGfuuu 
                 1366 
                 AAUGAAACAGAAAUA 
                 1488 
               
               
                   
                 AfAfAfuaaacuucs 
                   
                 auuuCfuGfuuucas 
                   
                 AACUUCCU 
                   
               
               
                   
                 csa 
                   
                 usu 
                   
                   
                   
               
               
                   
               
               
                 AD-1231062 
                 csuscaaa(Chd)aA 
                 1248 
                 VPusAfsucgUfgag 
                 1367 
                 UGCUCAAACAAGCAC 
                 1489 
               
               
                   
                 fGfCfacucacgasu 
                   
                 ugcuUfgUfuugags 
                   
                 UCACGAUU 
                   
               
               
                   
                 sa 
                   
                 csa 
                   
                   
                   
               
               
                   
               
               
                 AD-1231063 
                 asasacaaGfcAfCf 
                 1249 
                 VPusAfscaaUfcgu 
                 1368 
                 UCAAACAAGCACUCA 
                 1490 
               
               
                   
                 Ufcacga(Uhd)ugs 
                   
                 gaguGfcUfuguuus 
                   
                 CGAUUGUU 
                   
               
               
                   
                 usa 
                   
                 gsa 
                   
                   
                   
               
               
                   
               
               
                 AD-1231064 
                 asascaag(Chd)aC 
                 1250 
                 VPusAfsacaAfucg 
                 1369 
                 CAAACAAGCACUCAC 
                 1491 
               
               
                   
                 fUfCfacgauugusu 
                   
                 ugagUfgCfuuguus 
                   
                 GAUUGUUG 
                   
               
               
                   
                 sa 
                   
                 usg 
                   
                   
                   
               
               
                   
               
               
                 AD-1231074 
                 csusagca(Uhd)uG 
                 1251 
                 VPusAfscaaCfauu 
                 1370 
                 CCCUAGCAUUGGAAA 
                 1492 
               
               
                   
                 fGfAfaaauguugsu 
                   
                 uuccAfaUfgcuags 
                   
                 AUGUUGUA 
                   
               
               
                   
                 sa 
                   
                 gsg 
                   
                   
                   
               
               
                   
               
               
                 AD-1231078 
                 cscsagaa(Chd)aA 
                 1252 
                 VPusCfsaaaAfgaa 
                 1371 
                 GACCAGAACAAGAAU 
                 1493 
               
               
                   
                 fGfAfauucuuuusg 
                   
                 uucuUfgUfucuggs 
                   
                 UCUUUUGU 
                   
               
               
                   
                 sa 
                   
                 usc 
                   
                   
                   
               
               
                   
               
               
                 AD-1231079 
                 csasgaa(Chd)Afa 
                 1253 
                 VPusAfscaaAfaga 
                 1372 
                 ACCAGAACAAGAAUU 
                 1494 
               
               
                   
                 GfAfAfuucuuuugs 
                   
                 auucUfuGfuucugs 
                   
                 CUUUUGUG 
                   
               
               
                   
                 usa 
                   
                 gsu 
                   
                   
                   
               
               
                   
               
               
                 AD-1231084 
                 csusagggAfaAfGf 
                 1254 
                 VPusCfsacuGfaau 
                 1373 
                 GUCUAGGGAAAGUCA 
                 1495 
               
               
                   
                 Ufcauu(Chd)agus 
                   
                 gacuUfuCfccuags 
                   
                 UUCAGUGG 
                   
               
               
                   
                 gsa 
                   
                 asc 
                   
                   
                   
               
               
                   
               
               
                 AD-1231086 
                 usgscac(Chd)Ufa 
                 1255 
                 VPusCfsagaCfaca 
                 1374 
                 ACUGCACCUAAAAAU 
                 1496 
               
               
                   
                 AfAfAfaugugucus 
                   
                 uuuuUfaGfgugcas 
                   
                 GUGUCUGA 
                   
               
               
                   
                 gsa 
                   
                 gsu 
                   
                   
                   
               
               
                   
               
               
                 AD-1231088 
                 gsusau(Chd)aAfu 
                 1256 
                 VPusAfscggAfaag 
                 1375 
                 CCGUAUCAAUGAUGC 
                 1497 
               
               
                   
                 GfAfUfgcuuuccgs 
                   
                 caucAfuUfgauacs 
                   
                 UUUCCGUC 
                   
               
               
                   
                 usa 
                   
                 gsg 
                   
                   
                   
               
               
                   
               
               
                 AD-1231089 
                 asgsaaaa(Uhd)aA 
                 1257 
                 VPusGfsgaaUfccu 
                 1376 
                 GGAGAAAAUAAUCCA 
                 1498 
               
               
                   
                 fUfCfcaggauucsc 
                   
                 ggauUfaUfuuucus 
                   
                 GGAUUCCA 
                   
               
               
                   
                 sa 
                   
                 csc 
                   
                   
                   
               
               
                   
               
               
                 AD-1231090 
                 asasuc(Chd)aGfg 
                 1258 
                 VPusGfsuguUfuug 
                 1377 
                 AUAAUCCAGGAUUCC 
                 1499 
               
               
                   
                 AfUfUfccaaaacas 
                   
                 gaauCfcUfggauus 
                   
                 AAAACACU 
                   
               
               
                   
                 csa 
                   
                 asu 
                   
                   
                   
               
               
                   
               
               
                 AD-1231094 
                 uscscuu(Uhd)Ufa 
                 1259 
                 VPusCfsauaGfauu 
                 1378 
                 CCUCCUUUUAGAAAA 
                 1500 
               
               
                   
                 GfAfAfaaaucuaus 
                   
                 uuucUfaAfaaggas 
                   
                 AUCUAUGU 
                   
               
               
                   
                 gsa 
                   
                 gsg 
                   
                   
                   
               
               
                   
               
               
                 AD-1231095 
                 cscsucu(Uhd)Gfa 
                 1260 
                 VPusAfsacaAfaau 
                 1379 
                 UUUCUCUUGAGGUGA 
                 1501 
               
               
                   
                 GfGfUfgauuuugus 
                   
                 caccUfcAfagaggs 
                   
                 UUUUGUUG 
                   
               
               
                   
                 usa 
                   
                 asa 
                   
                   
                   
               
               
                   
               
               
                 AD-1231096 
                 asgscau(Chd)Ufu 
                 1261 
                 VPusGfscaaUfguc 
                 1380 
                 AGAGCAUCUUCAUUG 
                 1502 
               
               
                   
                 CfAfUfugacauugs 
                   
                 aaugAfaGfaugcus 
                   
                 ACAUUGCU 
                   
               
               
                   
                 csa 
                   
                 csu 
                   
                   
                   
               
               
                   
               
               
                 AD-1231097 
                 gscsauc(Uhd)Ufc 
                 1262 
                 VPusAfsgcaAfugu 
                 1381 
                 GAGCAUCUUCAUUGA 
                 1503 
               
               
                   
                 AfUfUfgacauugcs 
                   
                 caauGfaAfgaugcs 
                   
                 CAUUGCUU 
                   
               
               
                   
                 usa 
                   
                 usc 
                   
                   
                   
               
               
                   
               
               
                 AD-1231105 
                 ususauu(Uhd)Cfu 
                 1263 
                 VPusAfsaauCfcag 
                 1382 
                 AUUUAUUUCUGUCUC 
                 1504 
               
               
                   
                 GfUfCfucuggauus 
                   
                 agacAfgAfaauaas 
                   
                 UGGAUUUG 
                   
               
               
                   
                 usa 
                   
                 asu 
                   
                   
                   
               
               
                   
               
               
                 AD-1231106 
                 ususucug(Uhd)cU 
                 1264 
                 VPusGfsucaAfauc 
                 1383 
                 UAUUUCUGUCUCUGG 
                 1505 
               
               
                   
                 fCfUfggauuugasc 
                   
                 cagaGfaCfagaaas 
                   
                 AUUUGACU 
                   
               
               
                   
                 sa 
                   
                 usa 
                   
                   
                   
               
               
                   
               
               
                 AD-1231107 
                 uscsuc(Uhd)gGfa 
                 1265 
                 VPusAfscagAfagu 
                 1384 
                 UGUCUCUGGAUUUGA 
                 1506 
               
               
                   
                 UfUfUfgacuucugs 
                   
                 caaaUfcCfagagas 
                   
                 CUUCUGUU 
                   
               
               
                   
                 usa 
                   
                 csa 
                   
                   
                   
               
               
                   
               
               
                 AD-1231109 
                 usgsuu(Chd)aGfg 
                 1266 
                 VPusAfsuuuAfgau 
                 1385 
                 GCUGUUCAGGGAUAA 
                 1507 
               
               
                   
                 GfAfUfaaucuaaas 
                   
                 uaucCfcUfgaacas 
                   
                 UCUAAAUG 
                   
               
               
                   
                 usa 
                   
                 gsc 
                   
                   
                   
               
               
                   
               
               
                 AD-1231110 
                 asasaug(Uhd)Afa 
                 1267 
                 VPusUfsucaAfcag 
                 1386 
                 CUAAAUGUAAAUGUC 
                 1508 
               
               
                   
                 AfUfGfucuguugas 
                   
                 acauUfuAfcauuus 
                   
                 UGUUGAAU 
                   
               
               
                   
                 asa 
                   
                 asg 
                   
                   
                   
               
               
                   
               
               
                 AD-1231111 
                 asasugu(Chd)Ufg 
                 1268 
                 VPusUfscagAfaau 
                 1387 
                 UAAAUGUCUGUUGAA 
                 1509 
               
               
                   
                 UfUfGfaauuucugs 
                   
                 ucaaCfaGfacauus 
                   
                 UUUCUGAA 
                   
               
               
                   
                 asa 
                   
                 usa 
                   
                   
                   
               
               
                   
               
               
                 AD-1231112 
                 uscsaag(Uhd)Afc 
                 1269 
                 VPusCfsaaaUfcac 
                 1388 
                 AUCCAAGUACUAUGG 
                 1510 
               
               
                   
                 UfAfUfggugauuus 
                   
                 cauaGfuAfcuugas 
                   
                 UGAUUUGC 
                   
               
               
                   
                 gsa 
                   
                 asc 
                   
                   
                   
               
               
                   
               
               
                 AD-1231113 
                 csasagua(Chd)uA 
                 1270 
                 VPusGfscaaAfuca 
                 1389 
                 UCCAAGUACUAUGGU 
                 1511 
               
               
                   
                 fUfGfgugauuugsc 
                   
                 ccauAfgUfacuugs 
                   
                 GAUUUGCC 
                   
               
               
                   
                 sa 
                   
                 asa 
                   
                   
                   
               
               
                   
               
               
                 AD-1231114 
                 ususcaagGfuGfAf 
                 1271 
                 VPusUfsuuaGfacc 
                 1390 
                 UCUUCAAGGUGACAG 
                 1512 
               
               
                   
                 Cfaggu(Chd)uaas 
                   
                 ugucAfcCfuugaas 
                   
                 GUCUAAAG 
                   
               
               
                   
                 asa 
                   
                 gsa 
                   
                   
                   
               
               
                   
               
               
                 AD-1231115 
                 gsasaau(Uhd)Cfa 
                 1272 
                 VPusAfscugUfgag 
                 1391 
                 AAGAAAUUCAGAAUC 
                 1513 
               
               
                   
                 GfAfAfucucacags 
                   
                 auucUfgAfauuucs 
                   
                 UCACAGUC 
                   
               
               
                   
                 usa 
                   
                 usu 
                   
                   
                   
               
               
                   
               
               
                 AD-1231116 
                 gsasgga(Chd)Afu 
                 1273 
                 VPusAfsgugAfaaa 
                 1392 
                 UAGAGGACAUUGCUU 
                 1514 
               
               
                   
                 UfGfCfuuuuucacs 
                   
                 agcaAfuGfuccucs 
                   
                 UUUCACUU 
                   
               
               
                   
                 usa 
                   
                 usa 
                   
                   
                   
               
               
                   
               
               
                 AD-1231117 
                 gsgsaca(Uhd)Ufg 
                 1274 
                 VPusGfsaagUfgaa 
                 1393 
                 GAGGACAUUGCUUUU 
                 1515 
               
               
                   
                 CfUfUfuuucacuus 
                   
                 aaagCfaAfuguccs 
                   
                 UCACUUCC 
                   
               
               
                   
                 csa 
                   
                 usc 
                   
                   
                   
               
               
                   
               
               
                 AD-1231119 
                 gsasgugaAfuGfGf 
                 1275 
                 VPusGfsuugGfaau 
                 1394 
                 UGGAGUGAAUGGAAA 
                 1516 
               
               
                   
                 Afaauu(Chd)caas 
                   
                 uuccAfuUfcacucs 
                   
                 UUCCAACU 
                   
               
               
                   
                 csa 
                   
                 csa 
                   
                   
                   
               
               
                   
               
               
                 AD-1231120 
                 usgsaa(Uhd)gGfa 
                 1276 
                 VPusAfscagUfugg 
                 1395 
                 AGUGAAUGGAAAUUC 
                 1517 
               
               
                   
                 AfAfUfuccaacugs 
                   
                 aauuUfcCfauucas 
                   
                 CAACUGUA 
                   
               
               
                   
                 usa 
                   
                 csu 
               
               
                   
               
            
           
         
       
     
     
       
         
           
               
             
               
                 TABLE 6 
               
             
            
               
                   
               
               
                 ACE2 Single Dose Screens in PHH cells 
               
            
           
           
               
               
               
               
               
               
               
            
               
                   
                 10 nM 
                   
                 1 nM 
                   
                 0.1 nM 
                   
               
               
                 Duplex ID 
                 Avg 
                 SD 
                 Avg 
                 SD 
                 Avg 
                 SD 
               
               
                   
               
            
           
           
               
               
               
               
               
               
               
            
               
                 AD-1231127.1 
                 91.7 
                 4.8 
                 123.9 
                 26.7 
                 112.1 
                 14.7 
               
               
                 AD-1230985.1 
                 45.0 
                 17.9 
                 33.8 
                 8.9 
                 59.4 
                 8.7 
               
               
                 AD-1231084.1 
                 43.2 
                 6.6 
                 50.02 
                 10.60 
                 50.96 
                 18.81 
               
               
                 AD-1231052.1 
                 37.6 
                 10.5 
                 38.86 
                 12.20 
                 89.48 
                 9.79 
               
               
                 AD-1230929.1 
                 26.0 
                 9.5 
                 29.6 
                 3.6 
                 72.5 
                 17.5 
               
               
                 AD-1230840.1 
                 22.1 
                 5.8 
                 28.24 
                 5.32 
                 39.65 
                 4.94 
               
               
                 AD-1230933.1 
                 20.7 
                 3.2 
                 35.0 
                 6.5 
                 48.4 
                 16.5 
               
               
                 AD-1230885.1 
                 18.5 
                 1.8 
                 35.63 
                 7.85 
                 85.80 
                 19.63 
               
               
                 AD-1230844.1 
                 14.8 
                 6.3 
                 26.66 
                 8.15 
                 44.44 
                 18.08 
               
               
                 AD-1230886.1 
                 20.5 
                 6.2 
                 36.68 
                 8.56 
                 34.09 
                 5.99 
               
               
                 AD-1230846.1 
                 17.6 
                 6.3 
                 39.36 
                 14.19 
                 56.14 
                 24.78 
               
               
                 AD-1230822.1 
                 12.6 
                 3.8 
                 26.15 
                 6.36 
                 41.24 
                 16.49 
               
               
                 AD-1231121.1 
                 45.1 
                 17.6 
                 60.2 
                 22.4 
                 72.5 
                 11.9 
               
               
                 AD-1230847.1 
                 16.7 
                 2.0 
                 31.83 
                 7.54 
                 37.55 
                 9.26 
               
               
                 AD-1230848.1 
                 28.0 
                 6.6 
                 42.50 
                 3.64 
                 52.39 
                 9.32 
               
               
                 AD-1231072.1 
                 33.3 
                 11.8 
                 25.61 
                 6.69 
                 37.32 
                 9.85 
               
               
                 AD-1230849.1 
                 13.6 
                 1.4 
                 29.18 
                 9.09 
                 36.35 
                 6.07 
               
               
                 AD-1231130.1 
                 31.8 
                 13.1 
                 30.4 
                 11.3 
                 64.9 
                 9.7 
               
               
                 AD-1230851.1 
                 22.2 
                 12.9 
                 22.62 
                 9.16 
                 42.30 
                 20.24 
               
               
                 AD-1230821.1 
                 13.8 
                 2.0 
                 30.36 
                 17.09 
                 41.92 
                 12.18 
               
               
                 AD-1231123.1 
                 43.3 
                 8.9 
                 67.2 
                 6.0 
                 80.2 
                 10.0 
               
               
                 AD-1230855.1 
                 15.9 
                 6.4 
                 24.86 
                 5.46 
                 29.42 
                 10.68 
               
               
                 AD-1230856.1 
                 15.9 
                 5.2 
                 24.35 
                 5.67 
                 25.79 
                 11.56 
               
               
                 AD-1230894.1 
                 10.8 
                 2.7 
                 30.24 
                 14.60 
                 26.97 
                 4.06 
               
               
                 AD-1230895.1 
                 13.0 
                 2.9 
                 40.53 
                 7.97 
                 44.36 
                 10.52 
               
               
                 AD-1231073.1 
                 92.4 
                 34.7 
                 65.44 
                 17.65 
                 91.16 
                 21.23 
               
               
                 AD-1230937.1 
                 25.6 
                 9.7 
                 28.1 
                 12.8 
                 50.3 
                 15.0 
               
               
                 AD-1230896.1 
                 12.3 
                 2.4 
                 25.67 
                 4.59 
                 34.07 
                 3.16 
               
               
                 AD-1231075.1 
                 43.5 
                 9.2 
                 45.95 
                 17.34 
                 68.66 
                 25.05 
               
               
                 AD-1230999.1 
                 13.6 
                 1.2 
                 15.6 
                 2.7 
                 26.2 
                 3.3 
               
               
                 AD-1231076.1 
                 23.7 
                 4.2 
                 25.05 
                 6.65 
                 33.76 
                 10.94 
               
               
                 AD-1230897.1 
                 15.7 
                 4.2 
                 33.01 
                 7.35 
                 66.23 
                 15.14 
               
               
                 AD-1230938.1 
                 17.1 
                 7.2 
                 21.6 
                 5.4 
                 47.1 
                 4.2 
               
               
                 AD-1231077.1 
                 29.6 
                 3.9 
                 53.32 
                 16.07 
                 62.22 
                 14.14 
               
               
                 AD-1231080.1 
                 31.9 
                 6.9 
                 35.91 
                 1.75 
                 51.75 
                 11.10 
               
               
                 AD-1230939.1 
                 17.9 
                 3.0 
                 36.6 
                 9.7 
                 54.5 
                 13.6 
               
               
                 AD-1231085.1 
                 52.2 
                 1.3 
                 43.14 
                 6.30 
                 52.43 
                 10.90 
               
               
                 AD-1230944.1 
                 24.3 
                 7.0 
                 28.3 
                 7.8 
                 38.5 
                 11.6 
               
               
                 AD-1230945.1 
                 31.1 
                 7.4 
                 31.6 
                 12.7 
                 53.5 
                 18.1 
               
               
                 AD-1231006.1 
                 15.6 
                 6.4 
                 18.01 
                 2.07 
                 33.95 
                 4.95 
               
               
                 AD-1231022.1 
                 18.1 
                 4.0 
                 18.20 
                 3.26 
                 26.19 
                 3.46 
               
               
                 AD-1230962.1 
                 22.5 
                 7.6 
                 19.5 
                 1.5 
                 39.9 
                 10.1 
               
               
                 AD-1230904.1 
                 12.4 
                 5.4 
                 28.93 
                 4.31 
                 38.96 
                 8.75 
               
               
                 AD-1231115.1 
                 13.7 
                 1.9 
                 24.3 
                 3.9 
                 34.5 
                 7.6 
               
               
                 AD-1230905.1 
                 12.8 
                 3.0 
                 27.79 
                 7.41 
                 36.09 
                 4.40 
               
               
                 AD-1230906.1 
                 11.4 
                 2.8 
                 29.48 
                 5.12 
                 34.31 
                 8.53 
               
               
                 AD-1230907.1 
                 10.9 
                 4.4 
                 30.65 
                 3.01 
                 47.90 
                 2.38 
               
               
                 AD-1231023.1 
                 23.5 
                 5.1 
                 30.01 
                 2.89 
                 52.25 
                 8.09 
               
               
                 AD-1230864.1 
                 30.4 
                 7.2 
                 58.73 
                 4.37 
                 86.93 
                 25.34 
               
               
                 AD-1230865.1 
                 16.5 
                 3.7 
                 35.22 
                 6.16 
                 65.04 
                 15.88 
               
               
                 AD-1230963.1 
                 22.2 
                 6.0 
                 17.8 
                 11.3 
                 47.4 
                 14.1 
               
               
                 AD-1231024.1 
                 35.0 
                 9.5 
                 34.57 
                 11.49 
                 46.99 
                 3.10 
               
               
                 AD-1231033.1 
                 42.2 
                 18.4 
                 31.71 
                 5.49 
                 47.94 
                 8.30 
               
               
                 AD-1231034.1 
                 21.7 
                 5.9 
                 31.33 
                 9.03 
                 46.71 
                 3.92 
               
               
                 AD-1230969.1 
                 16.9 
                 7.4 
                 14.3 
                 2.5 
                 28.1 
                 5.3 
               
               
                 AD-1230867.1 
                 17.5 
                 7.6 
                 49.83 
                 8.80 
                 84.77 
                 27.19 
               
               
                 AD-1230868.1 
                 20.9 
                 7.6 
                 54.97 
                 6.47 
                 104.84 
                 30.54 
               
               
                 AD-1230869.1 
                 11.0 
                 6.4 
                 31.21 
                 11.40 
                 50.98 
                 10.86 
               
               
                 AD-1230970.1 
                 16.6 
                 7.7 
                 17.9 
                 5.1 
                 48.3 
                 20.8 
               
               
                 AD-1230971.1 
                 11.0 
                 3.6 
                 17.8 
                 7.7 
                 31.9 
                 4.6 
               
               
                 AD-1231035.1 
                 36.1 
                 13.6 
                 43.61 
                 8.31 
                 81.62 
                 6.73 
               
               
                 AD-1231122.1 
                 125.7 
                 27.3 
                 123.4 
                 14.1 
                 107.7 
                 24.0 
               
               
                 AD-1231132.1 
                 80.3 
                 11.2 
                 126.7 
                 18.7 
                 143.5 
                 27.5 
               
               
                 AD-1231124.1 
                 97.5 
                 21.3 
                 129.4 
                 12.8 
                 118.1 
                 15.2 
               
               
                 AD-1231125.1 
                 74.6 
                 5.6 
                 131.0 
                 36.5 
                 124.5 
                 21.5 
               
               
                 AD-1231036.1 
                 30.3 
                 13.4 
                 34.87 
                 4.93 
                 47.56 
                 4.22 
               
               
                 AD-1230972.1 
                 37.6 
                 11.9 
                 40.3 
                 27.5 
                 44.3 
                 22.0 
               
               
                 AD-1230973.1 
                 36.3 
                 12.3 
                 36.1 
                 16.8 
                 51.3 
                 5.7 
               
               
                 AD-1231037.1 
                 37.9 
                 7.4 
                 38.70 
                 4.38 
                 79.72 
                 18.31 
               
               
                 AD-1231038.1 
                 17.7 
                 5.4 
                 21.38 
                 2.59 
                 42.13 
                 1.18 
               
               
                 AD-1231039.1 
                 39.5 
                 15.0 
                 38.65 
                 8.60 
                 47.92 
                 3.57 
               
               
                 AD-1230974.1 
                 20.8 
                 6.7 
                 21.9 
                 6.9 
                 45.1 
                 9.2 
               
               
                 AD-1230909.1 
                 11.4 
                 2.1 
                 30.31 
                 7.17 
                 42.14 
                 10.01 
               
               
                 AD-1231040.1 
                 24.3 
                 9.4 
                 24.92 
                 7.62 
                 37.59 
                 3.57 
               
               
                 AD-1231041.1 
                 45.2 
                 13.2 
                 32.90 
                 11.10 
                 41.70 
                 7.45 
               
               
                 AD-1231042.1 
                 31.8 
                 10.3 
                 28.90 
                 3.61 
                 60.13 
                 15.27 
               
               
                 AD-1230975.1 
                 31.0 
                 6.4 
                 22.6 
                 9.7 
                 58.2 
                 13.7 
               
               
                 AD-1230976.1 
                 30.5 
                 9.2 
                 27.9 
                 7.7 
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                 14.3 
               
               
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                 54.1 
                 6.5 
                 39.36 
                 7.07 
                 91.91 
                 21.21 
               
               
                 AD-1231044.1 
                 18.8 
                 3.7 
                 23.76 
                 5.11 
                 72.60 
                 12.46 
               
               
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                 21.9 
                 5.9 
                 37.2 
                 11.8 
                 70.5 
                 13.7 
               
               
                 AD-1230978.1 
                 15.9 
                 9.8 
                 12.9 
                 6.0 
                 41.9 
                 9.1 
               
               
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                 14.0 
                 5.1 
                 30.19 
                 10.67 
                 36.07 
                 3.25 
               
               
                 AD-1230870.1 
                 15.2 
                 7.3 
                 36.95 
                 15.37 
                 77.89 
                 13.67 
               
               
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                 8.1 
                 2.3 
                 32.26 
                 5.38 
                 40.22 
                 5.49 
               
               
                 AD-1230979.1 
                 36.6 
                 16.5 
                 73.8 
                 23.5 
                 41.8 
                 21.8 
               
               
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                 19.0 
                 5.6 
                 19.85 
                 4.92 
                 45.80 
                 1.05 
               
               
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                 21.8 
                 10.4 
                 34.11 
                 1.99 
                 34.40 
                 10.89 
               
               
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                 14.2 
                 5.0 
                 25.98 
                 7.63 
                 51.38 
                 17.24 
               
               
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                 23.0 
                 4.0 
                 27.7 
                 15.1 
                 32.9 
                 21.8 
               
               
                 AD-1230912.1 
                 28.3 
                 13.5 
                 27.8 
                 13.2 
                 40.4 
                 12.1 
               
               
                 AD-1230980.1 
                 19.0 
                 6.3 
                 42.9 
                 10.1 
                 42.9 
                 6.9 
               
               
                 AD-1230981.1 
                 51.6 
                 8.6 
                 41.7 
                 5.0 
                 66.6 
                 9.2 
               
               
                 AD-1230982.1 
                 20.8 
                 6.8 
                 20.8 
                 2.1 
                 39.0 
                 5.5 
               
               
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                 37.4 
                 6.6 
                 53.8 
                 4.1 
                 94.9 
                 27.0 
               
               
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                 15.6 
                 1.1 
                 42.83 
                 8.17 
                 50.72 
                 9.89 
               
               
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                 38.7 
                 9.2 
                 42.6 
                 11.7 
                 48.9 
                 8.6 
               
               
                 AD-1230984.1 
                 72.6 
                 15.6 
                 64.5 
                 9.9 
                 98.1 
                 17.1 
               
               
                 AD-1231047.1 
                 34.7 
                 6.2 
                 47.28 
                 12.79 
                 76.03 
                 14.80 
               
               
                 AD-1231048.1 
                 47.4 
                 20.0 
                 39.55 
                 6.58 
                 84.57 
                 8.18 
               
               
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                 129.1 
                 30.0 
                 168.1 
                 30.4 
                 169.5 
                 36.2 
               
               
                 AD-1230823.1 
                 14.0 
                 4.3 
                 36.01 
                 4.75 
                 41.80 
                 8.42 
               
               
                 AD-1230871.1 
                 54.9 
                 16.9 
                 90.10 
                 16.36 
                 151.29 
                 11.49 
               
               
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                 93.7 
                 8.8 
                 95.5 
                 27.9 
                 86.1 
                 2.8 
               
               
                 AD-1230915.1 
                 24.9 
                 10.7 
                 21.6 
                 2.7 
                 57.7 
                 6.5 
               
               
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                 31.9 
                 4.1 
                 34.59 
                 5.98 
                 64.69 
                 13.14 
               
               
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                 28.8 
                 5.1 
                 44.4 
                 7.4 
                 38.6 
                 14.8 
               
               
                 AD-1230987.1 
                 51.1 
                 14.4 
                 52.7 
                 2.9 
                 54.7 
                 22.6 
               
               
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                 73.5 
                 4.4 
                 65.43 
                 17.25 
                 134.37 
                 34.95 
               
               
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                 38.6 
                 11.6 
                 73.6 
                 34.9 
                 80.2 
                 15.1 
               
               
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                 33.0 
                 7.9 
                 56.4 
                 7.6 
                 70.0 
                 5.0 
               
               
                 AD-1230916.1 
                 34.0 
                 13.5 
                 20.7 
                 8.0 
                 63.6 
                 11.7 
               
               
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                 28.6 
                 3.8 
                 33.3 
                 7.6 
                 64.0 
                 16.2 
               
               
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                 26.4 
                 12.5 
                 15.0 
                 4.7 
                 43.3 
                 11.5 
               
               
                 AD-1230834.1 
                 18.4 
                 3.4 
                 47.20 
                 10.25 
                 61.24 
                 25.59 
               
               
                 AD-1230872.1 
                 22.5 
                 10.5 
                 40.43 
                 8.64 
                 78.16 
                 13.70 
               
               
                 AD-1230991.1 
                 23.7 
                 3.8 
                 44.0 
                 17.0 
                 59.1 
                 11.3 
               
               
                 AD-1230918.1 
                 14.3 
                 5.3 
                 17.9 
                 7.9 
                 28.0 
                 4.9 
               
               
                 AD-1230873.1 
                 36.6 
                 24.5 
                 110.38 
                 13.27 
                 106.72 
                 12.39 
               
               
                 AD-1230919.1 
                 35.7 
                 14.3 
                 41.1 
                 5.4 
                 65.3 
                 13.1 
               
               
                 AD-1230874.1 
                 34.8 
                 24.8 
                 61.84 
                 20.74 
                 106.76 
                 35.91 
               
               
                 AD-1231051.1 
                 37.6 
                 10.6 
                 32.94 
                 5.69 
                 84.85 
                 23.12 
               
               
                 AD-1230992.1 
                 22.3 
                 3.7 
                 24.8 
                 8.8 
                 63.0 
                 5.8 
               
               
                 AD-1231053.1 
                 17.0 
                 6.4 
                 23.90 
                 4.23 
                 49.74 
                 12.41 
               
               
                 AD-1231054.1 
                 16.6 
                 12.2 
                 28.62 
                 2.33 
                 30.28 
                 10.68 
               
               
                 AD-1231055.1 
                 28.2 
                 9.8 
                 34.01 
                 10.21 
                 43.74 
                 9.27 
               
               
                 AD-1231056.1 
                 52.7 
                 18.2 
                 28.43 
                 7.84 
                 59.34 
                 2.82 
               
               
                 AD-1230875.1 
                 12.3 
                 7.7 
                 34.92 
                 3.34 
                 74.79 
                 24.31 
               
               
                 AD-1230876.1 
                 19.3 
                 6.1 
                 53.47 
                 18.20 
                 93.37 
                 10.37 
               
               
                 AD-1231057.1 
                 26.5 
                 10.4 
                 28.41 
                 0.98 
                 72.67 
                 14.83 
               
               
                 AD-1230835.1 
                 19.1 
                 6.0 
                 44.59 
                 8.06 
                 59.82 
                 3.58 
               
               
                 AD-1230824.1 
                 11.3 
                 3.3 
                 39.48 
                 2.81 
                 64.95 
                 10.14 
               
               
                 AD-1231058.1 
                 54.7 
                 30.2 
                 55.14 
                 12.32 
                 89.63 
                 29.24 
               
               
                 AD-1230920.1 
                 38.7 
                 12.6 
                 42.1 
                 2.7 
                 59.5 
                 10.2 
               
               
                 AD-1230993.1 
                 77.6 
                 13.4 
                 70.3 
                 15.0 
                 79.9 
                 14.0 
               
               
                 AD-1230921.1 
                 24.9 
                 5.0 
                 52.9 
                 19.3 
                 64.6 
                 8.7 
               
               
                 AD-1230922.1 
                 17.0 
                 1.6 
                 20.4 
                 3.7 
                 53.4 
                 9.5 
               
               
                 AD-1230923.1 
                 22.1 
                 8.4 
                 35.4 
                 17.4 
                 52.6 
                 8.4 
               
               
                 AD-1230994.1 
                 15.9 
                 2.9 
                 21.4 
                 4.1 
                 42.4 
                 14.7 
               
               
                 AD-1231059.1 
                 15.6 
                 7.9 
                 22.14 
                 12.55 
                 49.27 
                 7.82 
               
               
                 AD-1231060.1 
                 19.2 
                 5.4 
                 19.25 
                 8.99 
                 36.86 
                 9.41 
               
               
                 AD-1231061.1 
                 10.7 
                 5.0 
                 18.30 
                 2.60 
                 31.32 
                 19.52 
               
               
                 AD-1230924.1 
                 18.9 
                 16.1 
                 24.0 
                 9.8 
                 31.6 
                 9.3 
               
               
                 AD-1231062.1 
                 13.7 
                 5.8 
                 20.15 
                 3.19 
                 34.03 
                 12.00 
               
               
                 AD-1230925.1 
                 12.9 
                 6.4 
                 37.0 
                 18.1 
                 17.5 
                 3.8 
               
               
                 AD-1231063.1 
                 114.9 
                 16.8 
                 73.55 
                 10.83 
                 98.09 
                 16.11 
               
               
                 AD-1231064.1 
                 19.6 
                 8.4 
                 16.39 
                 3.18 
                 47.59 
                 15.88 
               
               
                 AD-1230926.1 
                 23.6 
                 7.3 
                 20.3 
                 7.2 
                 49.7 
                 4.6 
               
               
                 AD-1230927.1 
                 16.9 
                 2.6 
                 15.7 
                 3.0 
                 27.0 
                 7.1 
               
               
                 AD-1230995.1 
                 13.6 
                 1.8 
                 16.9 
                 4.8 
                 31.2 
                 4.7 
               
               
                 AD-1230928.1 
                 32.0 
                 9.2 
                 42.2 
                 13.5 
                 54.8 
                 7.8 
               
               
                 AD-1231065.1 
                 34.9 
                 5.8 
                 41.52 
                 10.52 
                 72.36 
                 13.62 
               
               
                 AD-1230930.1 
                 14.5 
                 5.5 
                 18.3 
                 13.4 
                 26.4 
                 12.4 
               
               
                 AD-1230836.1 
                 4.7 
                 2.1 
                 14.32 
                 2.68 
                 17.22 
                 5.11 
               
               
                 AD-1230931.1 
                 17.1 
                 7.5 
                 22.7 
                 7.2 
                 41.9 
                 5.1 
               
               
                 AD-1230837.1 
                 12.4 
                 5.6 
                 40.09 
                 3.80 
                 37.45 
                 10.14 
               
               
                 AD-1230877.1 
                 11.2 
                 4.3 
                 22.97 
                 3.56 
                 41.88 
                 12.37 
               
               
                 AD-1230878.1 
                 12.7 
                 5.9 
                 24.60 
                 5.67 
                 64.61 
                 7.91 
               
               
                 AD-1230879.1 
                 13.9 
                 2.5 
                 30.37 
                 5.01 
                 92.81 
                 6.97 
               
               
                 AD-1230838.1 
                 11.8 
                 7.8 
                 17.24 
                 3.46 
                 34.66 
                 10.88 
               
               
                 AD-1230839.1 
                 14.8 
                 5.9 
                 19.25 
                 6.73 
                 36.17 
                 13.54 
               
               
                 AD-1230932.1 
                 9.8 
                 5.6 
                 17.3 
                 13.0 
                 24.0 
                 4.0 
               
               
                 AD-1230996.1 
                 13.7 
                 6.0 
                 14.8 
                 5.4 
                 30.4 
                 7.5 
               
               
                 AD-1231066.1 
                 18.8 
                 6.9 
                 29.03 
                 11.10 
                 52.56 
                 16.33 
               
               
                 AD-1230880.1 
                 23.4 
                 14.5 
                 50.10 
                 8.52 
                 101.98 
                 39.70 
               
               
                 AD-1230841.1 
                 14.6 
                 3.7 
                 23.59 
                 5.39 
                 30.60 
                 7.30 
               
               
                 AD-1230881.1 
                 14.4 
                 5.9 
                 28.78 
                 7.69 
                 56.30 
                 17.95 
               
               
                 AD-1231067.1 
                 17.9 
                 5.0 
                 20.50 
                 6.42 
                 35.16 
                 6.81 
               
               
                 AD-1230934.1 
                 13.8 
                 4.9 
                 8.5 
                 7.4 
                 16.1 
                 15.0 
               
               
                 AD-1230842.1 
                 8.4 
                 3.5 
                 18.96 
                 4.46 
                 30.09 
                 4.58 
               
               
                 AD-1230882.1 
                 9.8 
                 3.2 
                 25.44 
                 5.68 
                 44.74 
                 8.92 
               
               
                 AD-1230825.1 
                 4.0 
                 1.3 
                 14.06 
                 7.33 
                 25.07 
                 10.26 
               
               
                 AD-1230825.2 
                 6.2 
                 1.7 
                 13.2 
                 2.3 
                 26.7 
                 4.1 
               
               
                 AD-1230826.1 
                 17.3 
                 1.8 
                 37.09 
                 10.75 
                 44.17 
                 8.31 
               
               
                 AD-1230843.1 
                 4.6 
                 1.2 
                 12.71 
                 5.54 
                 23.52 
                 5.33 
               
               
                 AD-1230883.1 
                 19.8 
                 9.8 
                 37.45 
                 8.76 
                 77.08 
                 11.14 
               
               
                 AD-1230884.1 
                 7.4 
                 1.3 
                 25.25 
                 2.52 
                 53.50 
                 5.67 
               
               
                 AD-1230845.1 
                 17.9 
                 5.0 
                 29.05 
                 7.12 
                 42.53 
                 22.22 
               
               
                 AD-1230887.1 
                 6.4 
                 1.7 
                 26.74 
                 5.91 
                 53.10 
                 14.78 
               
               
                 AD-1230935.1 
                 25.6 
                 5.2 
                 31.2 
                 23.4 
                 42.7 
                 3.4 
               
               
                 AD-1230888.1 
                 8.0 
                 3.4 
                 33.13 
                 9.60 
                 61.06 
                 21.41 
               
               
                 AD-1231068.1 
                 16.1 
                 3.7 
                 25.82 
                 7.95 
                 52.41 
                 15.27 
               
               
                 AD-1230997.1 
                 11.6 
                 3.1 
                 12.4 
                 3.7 
                 23.7 
                 4.4 
               
               
                 AD-1231069.1 
                 17.8 
                 2.6 
                 20.82 
                 4.94 
                 28.51 
                 5.82 
               
               
                 AD-1231070.1 
                 23.9 
                 8.9 
                 28.41 
                 7.11 
                 32.76 
                 6.12 
               
               
                 AD-1230889.1 
                 11.8 
                 3.0 
                 29.87 
                 13.43 
                 43.63 
                 13.38 
               
               
                 AD-1231071.1 
                 23.8 
                 4.4 
                 31.94 
                 9.06 
                 58.09 
                 4.45 
               
               
                 AD-1230998.1 
                 9.9 
                 2.6 
                 12.2 
                 2.3 
                 26.2 
                 6.1 
               
               
                 AD-1231131.1 
                 82.7 
                 12.4 
                 127.3 
                 10.7 
                 164.9 
                 84.4 
               
               
                 AD-1230936.1 
                 21.3 
                 7.7 
                 41.3 
                 10.6 
                 53.2 
                 4.2 
               
               
                 AD-1230890.1 
                 13.4 
                 7.9 
                 33.49 
                 3.24 
                 74.87 
                 12.91 
               
               
                 AD-1230850.1 
                 5.7 
                 2.5 
                 20.89 
                 5.67 
                 32.64 
                 6.93 
               
               
                 AD-1230827.1 
                 11.1 
                 4.7 
                 24.57 
                 6.91 
                 41.49 
                 13.64 
               
               
                 AD-1230828.1 
                 4.8 
                 1.6 
                 16.27 
                 2.68 
                 20.19 
                 3.88 
               
               
                 AD-1230852.1 
                 26.0 
                 12.1 
                 31.56 
                 2.89 
                 76.34 
                 27.94 
               
               
                 AD-1230891.1 
                 26.6 
                 11.5 
                 49.48 
                 16.60 
                 101.35 
                 10.03 
               
               
                 AD-1230892.1 
                 15.8 
                 5.9 
                 38.41 
                 9.59 
                 52.74 
                 21.72 
               
               
                 AD-1230853.1 
                 27.3 
                 9.4 
                 43.35 
                 12.63 
                 72.24 
                 23.92 
               
               
                 AD-1230854.1 
                 53.6 
                 8.2 
                 108.01 
                 9.95 
                 99.47 
                 26.40 
               
               
                 AD-1230893.1 
                 20.4 
                 8.0 
                 49.26 
                 6.40 
                 53.64 
                 3.29 
               
               
                 AD-1231074.1 
                 23.6 
                 6.8 
                 23.36 
                 8.79 
                 65.08 
                 6.02 
               
               
                 AD-1231000.1 
                 8.8 
                 2.4 
                 14.2 
                 4.0 
                 24.2 
                 6.3 
               
               
                 AD-1231078.1 
                 24.8 
                 7.7 
                 28.13 
                 8.95 
                 44.59 
                 7.53 
               
               
                 AD-1231079.1 
                 15.0 
                 5.3 
                 19.63 
                 7.84 
                 30.87 
                 5.31 
               
               
                 AD-1231081.1 
                 60.6 
                 20.9 
                 50.95 
                 13.92 
                 76.30 
                 19.18 
               
               
                 AD-1231082.1 
                 61.3 
                 14.6 
                 58.85 
                 6.33 
                 80.38 
                 4.31 
               
               
                 AD-1231083.1 
                 14.9 
                 5.9 
                 20.63 
                 2.27 
                 33.76 
                 10.47 
               
               
                 AD-1231001.1 
                 32.9 
                 9.3 
                 41.68 
                 25.80 
                 70.62 
                 23.58 
               
               
                 AD-1230940.1 
                 22.6 
                 10.7 
                 30.0 
                 19.6 
                 63.8 
                 8.1 
               
               
                 AD-1230829.1 
                 8.1 
                 4.2 
                 23.32 
                 7.59 
                 29.04 
                 10.58 
               
               
                 AD-1230941.1 
                 56.8 
                 16.3 
                 67.6 
                 36.2 
                 62.7 
                 1.1 
               
               
                 AD-1230857.1 
                 7.7 
                 1.2 
                 23.60 
                 4.19 
                 38.36 
                 12.48 
               
               
                 AD-1230858.1 
                 4.7 
                 2.6 
                 20.01 
                 3.05 
                 24.84 
                 6.65 
               
               
                 AD-1230898.1 
                 22.2 
                 13.0 
                 60.38 
                 15.85 
                 83.70 
                 28.45 
               
               
                 AD-1230899.1 
                 5.9 
                 1.7 
                 27.66 
                 8.37 
                 55.38 
                 10.01 
               
               
                 AD-1230900.1 
                 5.9 
                 1.8 
                 20.11 
                 2.88 
                 32.42 
                 5.58 
               
               
                 AD-1231142.1 
                 109.5 
                 27.5 
                 118.0 
                 10.4 
                 120.5 
                 29.5 
               
               
                 AD-1231128.1 
                 71.7 
                 12.1 
                 109.8 
                 11.8 
                 91.1 
                 24.6 
               
               
                 AD-1230901.1 
                 11.7 
                 6.7 
                 28.54 
                 2.48 
                 50.34 
                 15.63 
               
               
                 AD-1231086.1 
                 15.5 
                 4.5 
                 27.64 
                 7.84 
                 45.38 
                 9.97 
               
               
                 AD-1230942.1 
                 76.5 
                 9.9 
                 100.7 
                 40.7 
                 89.3 
                 11.0 
               
               
                 AD-1231002.1 
                 49.1 
                 5.6 
                 63.27 
                 9.13 
                 84.24 
                 12.61 
               
               
                 AD-1230859.1 
                 17.0 
                 4.5 
                 31.72 
                 2.92 
                 34.83 
                 5.08 
               
               
                 AD-1231003.1 
                 19.1 
                 9.7 
                 18.13 
                 6.23 
                 32.12 
                 2.47 
               
               
                 AD-1231087.1 
                 14.6 
                 3.4 
                 17.86 
                 5.16 
                 32.24 
                 10.06 
               
               
                 AD-1231004.1 
                 19.3 
                 5.4 
                 18.70 
                 5.83 
                 46.43 
                 10.03 
               
               
                 AD-1231088.1 
                 26.2 
                 3.6 
                 40.14 
                 4.72 
                 51.91 
                 5.45 
               
               
                 AD-1231005.1 
                 22.8 
                 7.3 
                 30.97 
                 4.25 
                 52.95 
                 14.15 
               
               
                 AD-1230943.1 
                 12.5 
                 6.0 
                 39.6 
                 19.1 
                 51.5 
                 13.3 
               
               
                 AD-1231090.1 
                 37.0 
                 6.7 
                 40.71 
                 2.70 
                 50.32 
                 10.97 
               
               
                 AD-1230946.1 
                 11.3 
                 2.3 
                 21.6 
                 8.0 
                 34.6 
                 5.2 
               
               
                 AD-1231092.1 
                 30.1 
                 5.4 
                 99.6 
                 12.6 
                 73.8 
                 15.6 
               
               
                 AD-1231093.1 
                 10.7 
                 4.0 
                 24.5 
                 10.7 
                 59.3 
                 13.6 
               
               
                 AD-1230947.1 
                 21.9 
                 7.1 
                 19.8 
                 7.3 
                 44.7 
                 6.9 
               
               
                 AD-1230948.1 
                 12.6 
                 5.2 
                 18.2 
                 5.1 
                 35.8 
                 3.8 
               
               
                 AD-1231007.1 
                 17.5 
                 4.3 
                 23.27 
                 6.84 
                 36.44 
                 7.58 
               
               
                 AD-1231094.1 
                 17.8 
                 4.0 
                 24.8 
                 5.2 
                 49.7 
                 3.2 
               
               
                 AD-1231095.1 
                 18.0 
                 3.4 
                 21.5 
                 3.8 
                 25.6 
                 4.4 
               
               
                 AD-1231008.1 
                 15.8 
                 2.9 
                 17.42 
                 5.15 
                 26.85 
                 11.05 
               
               
                 AD-1231096.1 
                 17.2 
                 3.8 
                 18.6 
                 3.1 
                 38.3 
                 12.6 
               
               
                 AD-1231097.1 
                 19.0 
                 2.6 
                 36.9 
                 9.4 
                 46.6 
                 12.9 
               
               
                 AD-1230830.1 
                 7.9 
                 1.8 
                 21.04 
                 7.33 
                 33.96 
                 15.64 
               
               
                 AD-1230949.1 
                 19.0 
                 14.8 
                 18.2 
                 18.4 
                 29.8 
                 5.3 
               
               
                 AD-1230950.1 
                 13.4 
                 3.6 
                 14.1 
                 2.5 
                 28.9 
                 9.8 
               
               
                 AD-1231009.1 
                 23.2 
                 4.3 
                 22.66 
                 7.75 
                 26.82 
                 4.35 
               
               
                 AD-1231098.1 
                 21.3 
                 10.6 
                 26.1 
                 10.1 
                 40.3 
                 12.1 
               
               
                 AD-1231010.1 
                 11.6 
                 2.1 
                 19.55 
                 5.04 
                 36.03 
                 10.54 
               
               
                 AD-1231099.1 
                 18.3 
                 4.8 
                 17.8 
                 4.8 
                 28.2 
                 10.9 
               
               
                 AD-1231011.1 
                 24.0 
                 5.0 
                 32.32 
                 9.21 
                 49.88 
                 7.94 
               
               
                 AD-1231100.1 
                 18.4 
                 6.8 
                 24.1 
                 6.4 
                 37.7 
                 13.8 
               
               
                 AD-1231101.1 
                 18.9 
                 2.9 
                 38.2 
                 8.6 
                 51.4 
                 17.8 
               
               
                 AD-1231102.1 
                 19.5 
                 1.2 
                 26.4 
                 6.3 
                 42.0 
                 7.9 
               
               
                 AD-1231103.1 
                 15.6 
                 2.5 
                 31.1 
                 9.3 
                 43.2 
                 12.9 
               
               
                 AD-1231104.1 
                 18.2 
                 6.4 
                 37.1 
                 7.0 
                 33.1 
                 10.5 
               
               
                 AD-1231012.1 
                 20.8 
                 4.6 
                 22.28 
                 4.49 
                 50.16 
                 11.63 
               
               
                 AD-1230951.1 
                 38.1 
                 18.8 
                 26.0 
                 4.4 
                 39.1 
                 10.1 
               
               
                 AD-1231105.1 
                 25.7 
                 0.5 
                 24.7 
                 7.9 
                 46.9 
                 9.4 
               
               
                 AD-1231106.1 
                 14.9 
                 7.9 
                 25.5 
                 3.1 
                 34.4 
                 7.0 
               
               
                 AD-1231013.1 
                 16.4 
                 3.1 
                 19.42 
                 1.16 
                 42.25 
                 13.12 
               
               
                 AD-1230952.1 
                 13.7 
                 6.0 
                 23.7 
                 9.5 
                 38.9 
                 6.6 
               
               
                 AD-1231107.1 
                 22.9 
                 2.5 
                 18.7 
                 7.9 
                 40.3 
                 14.4 
               
               
                 AD-1231014.1 
                 8.8 
                 0.7 
                 17.25 
                 2.99 
                 35.35 
                 3.12 
               
               
                 AD-1230953.1 
                 20.0 
                 7.0 
                 18.2 
                 3.3 
                 32.0 
                 4.7 
               
               
                 AD-1231015.1 
                 14.4 
                 1.2 
                 20.31 
                 3.14 
                 32.01 
                 15.32 
               
               
                 AD-1231016.1 
                 13.8 
                 1.0 
                 23.79 
                 3.56 
                 33.69 
                 7.05 
               
               
                 AD-1231017.1 
                 16.4 
                 1.8 
                 23.58 
                 5.23 
                 29.38 
                 11.43 
               
               
                 AD-1231129.1 
                 89.2 
                 14.2 
                 129.6 
                 15.2 
                 121.0 
                 26.9 
               
               
                 AD-1230954.1 
                 16.4 
                 6.3 
                 22.3 
                 6.4 
                 39.3 
                 11.7 
               
               
                 AD-1231108.1 
                 15.3 
                 2.7 
                 25.7 
                 8.4 
                 39.8 
                 6.4 
               
               
                 AD-1230902.1 
                 10.5 
                 3.4 
                 21.73 
                 3.16 
                 34.09 
                 4.81 
               
               
                 AD-1231109.1 
                 43.0 
                 6.2 
                 54.5 
                 12.8 
                 81.1 
                 25.4 
               
               
                 AD-1230955.1 
                 11.4 
                 5.0 
                 25.0 
                 4.5 
                 39.5 
                 7.7 
               
               
                 AD-1230956.1 
                 31.9 
                 10.1 
                 34.4 
                 20.7 
                 27.4 
                 23.3 
               
               
                 AD-1230957.1 
                 25.5 
                 7.3 
                 22.5 
                 11.2 
                 36.1 
                 11.5 
               
               
                 AD-1231110.1 
                 16.2 
                 4.6 
                 28.0 
                 6.4 
                 27.7 
                 9.0 
               
               
                 AD-1231018.1 
                 15.8 
                 8.0 
                 21.32 
                 7.80 
                 40.75 
                 8.52 
               
               
                 AD-1230860.1 
                 25.9 
                 9.4 
                 29.77 
                 15.13 
                 100.17 
                 34.11 
               
               
                 AD-1231019.1 
                 21.7 
                 2.9 
                 22.47 
                 4.04 
                 37.01 
                 7.04 
               
               
                 AD-1231111.1 
                 13.9 
                 3.0 
                 25.6 
                 5.7 
                 23.8 
                 4.0 
               
               
                 AD-1230958.1 
                 18.3 
                 11.7 
                 22.3 
                 9.2 
                 51.7 
                 16.0 
               
               
                 AD-1231020.1 
                 32.8 
                 9.0 
                 26.23 
                 5.77 
                 42.09 
                 9.94 
               
               
                 AD-1230959.1 
                 21.4 
                 7.9 
                 23.9 
                 6.1 
                 55.4 
                 11.5 
               
               
                 AD-1231021.1 
                 18.4 
                 4.6 
                 19.87 
                 6.31 
                 33.42 
                 5.79 
               
               
                 AD-1231133.1 
                 75.3 
                 13.6 
                 134.2 
                 19.5 
                 126.2 
                 27.6 
               
               
                 AD-1230861.1 
                 21.0 
                 4.3 
                 34.76 
                 15.77 
                 108.47 
                 16.83 
               
               
                 AD-1230960.1 
                 21.7 
                 9.6 
                 19.2 
                 5.1 
                 35.8 
                 7.8 
               
               
                 AD-1230903.1 
                 10.3 
                 1.4 
                 30.41 
                 3.26 
                 41.91 
                 19.64 
               
               
                 AD-1230862.1 
                 18.8 
                 7.3 
                 38.04 
                 10.54 
                 74.51 
                 27.01 
               
               
                 AD-1230961.1 
                 37.9 
                 10.4 
                 32.1 
                 6.9 
                 60.4 
                 11.7 
               
               
                 AD-1231141.1 
                 118.6 
                 24.2 
                 154.7 
                 29.6 
                 151.4 
                 37.4 
               
               
                 AD-1231134.1 
                 109.8 
                 25.6 
                 157.8 
                 40.8 
                 142.6 
                 14.8 
               
               
                 AD-1231135.1 
                 83.1 
                 29.9 
                 105.7 
                 26.2 
                 110.7 
                 18.8 
               
               
                 AD-1231112.1 
                 63.1 
                 7.4 
                 87.5 
                 6.5 
                 91.3 
                 15.9 
               
               
                 AD-1231113.1 
                 18.2 
                 3.2 
                 39.0 
                 9.5 
                 47.8 
                 5.8 
               
               
                 AD-1231136.1 
                 121.4 
                 8.9 
                 154.8 
                 38.2 
                 124.0 
                 13.5 
               
               
                 AD-1231114.1 
                 26.3 
                 10.8 
                 29.3 
                 14.0 
                 41.2 
                 10.3 
               
               
                 AD-1231116.1 
                 13.6 
                 1.2 
                 24.7 
                 10.6 
                 40.2 
                 8.0 
               
               
                 AD-1231117.1 
                 16.3 
                 5.4 
                 35.4 
                 10.1 
                 48.7 
                 21.1 
               
               
                 AD-1230863.1 
                 24.1 
                 15.5 
                 31.31 
                 5.43 
                 47.89 
                 12.23 
               
               
                 AD-1231138.1 
                 139.7 
                 50.3 
                 157.0 
                 36.2 
                 168.0 
                 66.9 
               
               
                 AD-1231139.1 
                 46.9 
                 3.8 
                 63.4 
                 6.5 
                 105.7 
                 31.8 
               
               
                 AD-1231140.1 
                 107.9 
                 27.0 
                 133.0 
                 25.6 
                 155.6 
                 17.9 
               
               
                 AD-1231118.1 
                 14.7 
                 4.5 
                 31.6 
                 5.7 
                 38.6 
                 2.1 
               
               
                 AD-1230908.1 
                 12.1 
                 2.6 
                 32.95 
                 3.83 
                 43.15 
                 9.69 
               
               
                 AD-1231126.1 
                 94.5 
                 10.5 
                 134.1 
                 18.0 
                 96.3 
                 35.2 
               
               
                 AD-1231025.1 
                 21.7 
                 2.4 
                 26.88 
                 8.92 
                 43.45 
                 8.81 
               
               
                 AD-1231119.1 
                 20.7 
                 4.5 
                 27.0 
                 4.2 
                 45.5 
                 8.9 
               
               
                 AD-1231120.1 
                 18.5 
                 4.1 
                 39.6 
                 14.0 
                 38.7 
                 10.9 
               
               
                 AD-1231026.1 
                 14.2 
                 1.8 
                 26.38 
                 7.85 
                 40.28 
                 13.98 
               
               
                 AD-1231027.1 
                 25.6 
                 11.3 
                 19.90 
                 2.21 
                 42.10 
                 11.73 
               
               
                 AD-1230964.1 
                 18.9 
                 5.6 
                 12.8 
                 7.5 
                 20.0 
                 8.7 
               
               
                 AD-1231028.1 
                 23.1 
                 3.3 
                 23.06 
                 4.74 
                 49.31 
                 10.19 
               
               
                 AD-1230866.1 
                 12.2 
                 3.2 
                 24.42 
                 7.65 
                 41.88 
                 5.69 
               
               
                 AD-1230965.1 
                 22.1 
                 10.6 
                 14.7 
                 6.4 
                 22.6 
                 3.9 
               
               
                 AD-1230966.1 
                 16.2 
                 4.9 
                 17.3 
                 1.5 
                 32.8 
                 13.2 
               
               
                 AD-1231143.1 
                 46.1 
                 38.8 
                 93.6 
                 18.1 
                 96.6 
                 21.4 
               
               
                 AD-1231029.1 
                 37.5 
                 10.4 
                 37.04 
                 9.59 
                 48.84 
                 10.67 
               
               
                 AD-1231030.1 
                 17.5 
                 2.4 
                 18.66 
                 2.70 
                 24.48 
                 12.30 
               
               
                 AD-1231031.1 
                 20.3 
                 8.3 
                 35.22 
                 10.71 
                 35.54 
                 10.28 
               
               
                 AD-1231032.1 
                 25.5 
                 3.8 
                 28.76 
                 8.44 
                 38.44 
                 5.06 
               
               
                 AD-1230967.1 
                 28.6 
                 10.8 
                 23.6 
                 4.8 
                 32.3 
                 8.9 
               
               
                 AD-1230968.1 
                 26.3 
                 14.0 
                 25.2 
                 6.7 
                 48.5 
                 17.5 
               
               
                   
               
            
           
         
       
     
     Example 3. In Vivo Screening of dsRNA Duplexes in Mice 
     siRNA molecules targeting the ACE2 gene, identified from the above in vitro studies, are evaluated in vivo. 
     Mice previously infected with a coronavirus, e.g., severe acute respiratory syndrome-2 (SARS-2)-CoV-2, are administered, via pulmonary or subcutaneous delivery, a dsRNA molecule at a dose of 0.1 mg/kg, 1 mg/kg or 10 mg/kg. Uptake of dsRNA in bronchioles and alveoli and expression level of target gene in whole lung of treated mice are measured. Expression levels of coronavirus target genes are further evaluated by in situ hybridization in mice bronchus and bronchiole. 
     Example 4. In Vivo Screening of dsRNA Duplexes in Mice 
     siRNA molecules targeting the ACE2 gene, identified from the above in vitro studies, were evaluated in vivo. 
     B6.Cg-Tg(K18-ACE2)2Prlmn/J transgenic mice (mice overexpressing the human ACE2 gene) were administered a single 10/kg dose via intranasal instillation in a total volume of 50 μL (25 μL/nostril) of duplex AD-1230825, AD-1230843, or AD-1230934, or PBS. At Day 10 post-dose, lungs were harvested and the level of human ACE2 activity was evaluated by RT-qPCR (N=3, per group). 
     As shown in Table 7, all of the intranasally instilled agents effectively and potently lowered human ACE-2 mRNA levels. 
     
       
         
           
               
             
               
                 TABLE 7 
               
             
            
               
                   
               
               
                 Human ACE-2 (hACE-2) % Transcript Remaning 
               
            
           
           
               
               
               
               
               
            
               
                 Animal # 
                 PBS 
                 AD-1230825 
                 AD-1230843 
                 AD-1230934 
               
               
                   
               
            
           
           
               
               
               
               
               
            
               
                 1 
                 104.49 
                 27.52 
                 23.79 
                 12.27 
               
               
                 2 
                 88.17 
                 27.33 
                 29.8 
                 44.85 
               
               
                 3 
                 108.55 
                 38.78 
                 46.28 
                 24.46 
               
               
                 Average 
                 100.4 
                 31.2 
                 33.3 
                 27.2 
               
               
                 Median 
                 104.5 
                 27.5 
                 29.8 
                 24.5 
               
               
                   
               
            
           
         
       
     
     Example 5. Intranasal Delivery of dsRNA Duplexes Prevents Coronavirus Infection 
     Experimental Design 
     To determine the efficacy of dsRNA agents administered intranasally, fifty-four (54) Male Syrian Golden hamsters, approximately 6-8 weeks of age were divided among seven groups, according to Table 10, below, in groups of 6 animals Group 1 was a control group administered PBS via intranasal (IN) dosing on day −7 pre-challenge. Group 2 was a control group administered a dsRNA agent targeting luciferase via intranasal (IN) dosing on day −7 pre-challenge. Groups 3-6 were administered either a combination of AD-1184150 and AD-1184137, both targeting COVID-19, or AD-1230934, targeting ACE2 (see Table 10) via intranasal (IN) dosing on day −7 pre-challenge. Group 7 was administered a combination of AD-1184150 and AD-1184137, via subcutaneous (SQ) dosing on day −7 pre-challenge. 
     Animals were challenged on study day 0 with SARS-CoV-2 via the intranasal route Animals were monitored to Day 7 post-challenge. Oral swabs were collected in the post-challenge period, days 1, 3, and 5. Terminal oral swabs, blood, and tissue collection occurred on day 7 post-challenge. 
     
       
         
           
               
             
               
                 TABLE 10 
               
             
            
               
                   
               
               
                 Experimental Design 
               
            
           
           
               
               
               
               
               
               
            
               
                   
                   
                   
                   
                 Dose at 
                 Treatment 
               
               
                 Group 
                 N 
                 Treatment 
                 Route 
                 each Treatment 
                 Days 
               
               
                   
               
            
           
           
               
               
               
               
               
               
            
               
                 1 
                 6 
                 PBS 
                 IN 
                  0 mg/kg 
                 SD −7 
               
               
                 2 
                 6 
                 Luciferase 
                 IN 
                 30 mg/kg 
                 SD −7 
               
               
                 3 
                 6 
                 AD-1184150 + 
                 IN 
                 30 mg/kg 
                 SD −7 
               
               
                   
                   
                 AD-1184137* 
               
               
                 4 
                 6 
                 AD-1230934 
                 IN 
                 30 mg/kg 
                 SD −7 
               
               
                 5 
                 6 
                 AD-1184150 + 
                 IN 
                 10 mg/kg 
                 SD −7 
               
               
                   
                   
                 AD-1184137 
               
               
                 6 
                 6 
                 AD-1184150 + 
                 IN 
                  1 mg/kg 
                 SD −7 
               
               
                   
                   
                 AD-1184137 
               
               
                 7 
                 5 
                 AD-1184150 + 
                 SQ 
                 30 mg/kg 
                 SD −7 
               
               
                   
                   
                 AD-1184137 
               
               
                   
               
               
                 *Described in patent application Attorney Docket No. 121301-12220 filed on Mar. 25, 2021, the entire contents of which are incorporated herein by reference. 
               
            
           
         
       
     
     For animals receiving a combination of AD-1184150 and AD-1184137, the two duplexes were mixed together and the weight administered to each animal, as indicated in Table 10, is the total weight of the mixture of the two duplexes. 
     Each animal received a dose volume for IN dosing of 100 μl per animal (50 μl per nostril) and 200 μl per animal for SQ dosing. 
     Virus Challenge with SARS-CoV-2 
     The intranasal inoculation (IN) was performed on Ketamine/Xylazine anesthetized hamsters. 
     Administration of virus was conducted as follows: using a calibrated P200 pipettor, 50 μL of the viral inoculum was administered dropwise into each nostril, for a total of 100 μL per animal. Anesthetized animals were held upright such that the nostrils of the hamster were pointing towards the ceiling. The tip of the syringe was placed into the first nostril and virus inoculum was slowly injecting into the nasal passage, and then removed. This was repeated for the second nostril. The animal&#39;s head was tilted back for about 20 seconds and then returned to its housing unit and monitored until fully recovered. 
     Body weights were determined each day post-challenge through Day 7 post-challenge to assess the effectiveness of the duplexes as assessed by the weight of the animals. 
     The results are provided in  FIG.  1    and demonstrate that intranasal administration of a single 30 mg/kg dose of AD-1230934 prevents SARS-CoV-2 infection as demonstrated by the maintenance of the weights of the hamsters. 
     EQUIVALENTS 
     Those skilled in the art will recognize, or be able to ascertain using no more than routine experimentation, many equivalents to the specific embodiments and methods described herein. Such equivalents are intended to be encompassed by the scope of the following claims. 
     
       
         
           
               
             
               
                   
               
               
                 INFORMAL SEQUENCE LISTING 
               
               
                   
               
             
            
               
                   
               
            
           
           
               
               
            
               
                 &lt;210&gt; 
                  1 
               
               
                 &lt;211&gt; 
                 3596 
               
               
                 &lt;212&gt; 
                 DNA 
               
               
                 &lt;213&gt;  
                 
                   Homo sapiens 
                 
               
               
                 &lt;400&gt; 
                  1 
               
            
           
           
               
               
            
               
                 ggcactcata catacactct ggcaatgagg acactgagct cgcttctgaa atttgacaag 
                   60 
               
               
                 ataaccacta aaatctcttt gaattctatg ttgttgtgat cccatggcta cagaggatca 
                  120 
               
               
                 ggagttgaca tagatactct ttggatttca taccatgtgg aggctttctt acttccacgt 
                  180 
               
               
                 gaccttgact gagttttgaa tagcgcccaa cccaagttca aaggctgata agagagaaaa 
                  240 
               
               
                 tctcatgagg aggttttagt ctagggaaag tcattcagtg gatgtgatct tggctcacag 
                  300 
               
               
                 gggacgatgt caagctcttc ctggctcctt ctcagccttg ttgctgtaac tgctgctcag 
                  360 
               
               
                 tccaccattg aggaacaggc caagacattt ttggacaagt ttaaccacga agccgaagac 
                  420 
               
               
                 ctgttctatc aaagttcact tgcttcttgg aattataaca ccaatattac tgaagagaat 
                  480 
               
               
                 gtccaaaaca tgaataatgc tggggacaaa tggtctgcct ttttaaagga acagtccaca 
                  540 
               
               
                 cttgcccaaa tgtatccact acaagaaatt cagaatctca cagtcaagct tcagctgcag 
                  600 
               
               
                 gctcttcagc aaaatgggtc ttcagtgctc tcagaagaca agagcaaacg gttgaacaca 
                  660 
               
               
                 attctaaata caatgagcac catctacagt actggaaaag tttgtaaccc agataatcca 
                  720 
               
               
                 caagaatgct tattacttga accaggtttg aatgaaataa tggcaaacag tttagactac 
                  780 
               
               
                 aatgagaggc tctgggcttg ggaaagctgg agatctgagg tcggcaagca gctgaggcca 
                  840 
               
               
                 ttatatgaag agtatgtggt cttgaaaaat gagatggcaa gagcaaatca ttatgaggac 
                  900 
               
               
                 tatggggatt attggagagg agactatgaa gtaaatgggg tagatggcta tgactacagc 
                  960 
               
               
                 cgcggccagt tgattgaaga tgtggaacat acctttgaag agattaaacc attatatgaa 
                 1020 
               
               
                 catcttcatg cctatgtgag ggcaaagttg atgaatgcct atccttccta tatcagtcca 
                 1080 
               
               
                 attggatgcc tccctgctca tttgcttggt gatatgtggg gtagattttg gacaaatctg 
                 1140 
               
               
                 tactctttga cagttccctt tggacagaaa ccaaacatag atgttactga tgcaatggtg 
                 1200 
               
               
                 gaccaggcct gggatgcaca gagaatattc aaggaggccg agaagttctt tgtatctgtt 
                 1260 
               
               
                 ggtcttccta atatgactca aggattctgg gaaaattcca tgctaacgga cccaggaaat 
                 1320 
               
               
                 gttcagaaag cagtctgcca tcccacagct tgggacctgg ggaagggcga cttcaggatc 
                 1380 
               
               
                 cttatgtgca caaaggtgac aatggacgac ttcctgacag ctcatcatga gatggggcat 
                 1440 
               
               
                 atccagtatg atatggcata tgctgcacaa ccttttctgc taagaaatgg agctaatgaa 
                 1500 
               
               
                 ggattccatg aagctgttgg ggaaatcatg tcactttctg cagccacacc taagcattta 
                 1560 
               
               
                 aaatccattg gtcttctgtc acccgatttt caagaagaca atgaaacaga aataaacttc 
                 1620 
               
               
                 ctgctcaaac aagcactcac gattgttggg actctgccat ttacttacat gttagagaag 
                 1680 
               
               
                 tggaggtgga tggtctttaa aggggaaatt cccaaagacc agtggatgaa aaagtggtgg 
                 1740 
               
               
                 gagatgaagc gagagatagt tggggtggtg gaacctgtgc cccatgatga aacatactgt 
                 1800 
               
               
                 gaccccgcat ctctgttcca tgtttctaat gattactcat tcattcgata ttacacaagg 
                 1860 
               
               
                 accctttacc aattccagtt tcaagaagca ctttgtcaag cagctaaaca tgaaggccct 
                 1920 
               
               
                 ctgcacaaat gtgacatctc aaactctaca gaagctggac agaaactgtt caatatgctg 
                 1980 
               
               
                 aggcttggaa aatcagaacc ctggacccta gcattggaaa atgttgtagg agcaaagaac 
                 2040 
               
               
                 atgaatgtaa ggccactgct caactacttt gagcccttat ttacctggct gaaagaccag 
                 2100 
               
               
                 aacaagaatt cttttgtggg atggagtacc gactggagtc catatgcaga ccaaagcatc 
                 2160 
               
               
                 aaagtgagga taagcctaaa atcagctctt ggagataaag catatgaatg gaacgacaat 
                 2220 
               
               
                 gaaatgtacc tgttccgatc atctgttgca tatgctatga ggcagtactt tttaaaagta 
                 2280 
               
               
                 aaaaatcaga tgattctttt tggggaggag gatgtgcgag tggctaattt gaaaccaaga 
                 2340 
               
               
                 atctccttta atttctttgt cactgcacct aaaaatgtgt ctgatatcat tcctagaact 
                 2400 
               
               
                 gaagttgaaa aggccatcag gatgtcccgg agccgtatca atgatgcttt ccgtctgaat 
                 2460 
               
               
                 gacaacagcc tagagtttct ggggatacag ccaacacttg gacctcctaa ccagccccct 
                 2520 
               
               
                 gtttccatat ggctgattgt ttttggagtt gtgatgggag tgatagtggt tggcattgtc 
                 2580 
               
               
                 atcctgatct tcactgggat cagagatcgg aagaagaaaa ataaagcaag aagtggagaa 
                 2640 
               
               
                 aatccttatg cctccatcga tattagcaaa ggagaaaata atccaggatt ccaaaacact 
                 2700 
               
               
                 gatgatgttc agacctcctt ttagaaaaat ctatgttttt cctcttgagg tgattttgtt 
                 2760 
               
               
                 gtatgtaaat gttaatttca tggtatagaa aatataagat gataaagata tcattaaatg 
                 2820 
               
               
                 tcaaaactat gactctgttc agaaaaaaaa ttgtccaaag acaacatggc caaggagaga 
                 2880 
               
               
                 gcatcttcat tgacattgct ttcagtattt atttctgtct ctggatttga cttctgttct 
                 2940 
               
               
                 gtttcttaat aaggattttg tattagagta tattagggaa agtgtgtatt tggtctcaca 
                 3000 
               
               
                 ggctgttcag ggataatcta aatgtaaatg tctgttgaat ttctgaagtt gaaaacaagg 
                 3060 
               
               
                 atatatcatt ggagcaagtg ttggatcttg tatggaatat ggatggatca cttgtaagga 
                 3120 
               
               
                 cagtgcctgg gaactggtgt agctgcaagg attgagaatg gcatgcatta gctcactttc 
                 3180 
               
               
                 atttaatcca ttgtcaagga tgacatgctt tcttcacagt aactcagttc aagtactatg 
                 3240 
               
               
                 gtgatttgcc tacagtgatg tttggaatcg atcatgcttt cttcaaggtg acaggtctaa 
                 3300 
               
               
                 agagagaaga atccagggaa caggtagagg acattgcttt ttcacttcca aggtgcttga 
                 3360 
               
               
                 tcaacatctc cctgacaaca caaaactaga gccaggggcc tccgtgaact cccagagcat 
                 3420 
               
               
                 gcctgataga aactcatttc tactgttctc taactgtgga gtgaatggaa attccaactg 
                 3480 
               
               
                 tatgttcacc ctctgaagtg ggtacccagt ctcttaaatc ttttgtattt gctcacagtg 
                 3540 
               
               
                 tttgagcagt gctgagcaca aagcagacac tcaataaatg ctagatttac acactc 
                 3596 
               
               
                   
               
            
           
           
               
               
            
               
                 &lt;210&gt; 
                  2 
               
               
                 &lt;211&gt; 
                 3339 
               
               
                 &lt;212&gt; 
                 DNA 
               
               
                 &lt;213&gt; 
                 
                   Homo sapiens 
                 
               
               
                 &lt;400&gt; 
                  2 
               
            
           
           
               
               
            
               
                 agtctaggga aagtcattca gtggatgtga tcttggctca caggggacga tgtcaagctc 
                   60 
               
               
                 ttcctggctc cttctcagcc ttgttgctgt aactgctgct cagtccacca ttgaggaaca 
                  120 
               
               
                 ggccaagaca tttttggaca agtttaacca cgaagccgaa gacctgttct atcaaagttc 
                  180 
               
               
                 acttgcttct tggaattata acaccaatat tactgaagag aatgtccaaa acatgaataa 
                  240 
               
               
                 tgctggggac aaatggtctg cctttttaaa ggaacagtcc acacttgccc aaatgtatcc 
                  300 
               
               
                 actacaagaa attcagaatc tcacagtcaa gcttcagctg caggctcttc agcaaaatgg 
                  360 
               
               
                 gtcttcagtg ctctcagaag acaagagcaa acggttgaac acaattctaa atacaatgag 
                  420 
               
               
                 caccatctac agtactggaa aagtttgtaa cccagataat ccacaagaat gcttattact 
                  480 
               
               
                 tgaaccaggt ttgaatgaaa taatggcaaa cagtttagac tacaatgaga ggctctgggc 
                  540 
               
               
                 ttgggaaagc tggagatctg aggtcggcaa gcagctgagg ccattatatg aagagtatgt 
                  600 
               
               
                 ggtcttgaaa aatgagatgg caagagcaaa tcattatgag gactatgggg attattggag 
                  660 
               
               
                 aggagactat gaagtaaatg gggtagatgg ctatgactac agccgcggcc agttgattga 
                  720 
               
               
                 agatgtggaa catacctttg aagagattaa accattatat gaacatcttc atgcctatgt 
                  780 
               
               
                 gagggcaaag ttgatgaatg cctatccttc ctatatcagt ccaattggat gcctccctgc 
                  840 
               
               
                 tcatttgctt ggtgatatgt ggggtagatt ttggacaaat ctgtactctt tgacagttcc 
                  900 
               
               
                 ctttggacag aaaccaaaca tagatgttac tgatgcaatg gtggaccagg cctgggatgc 
                  960 
               
               
                 acagagaata ttcaaggagg ccgagaagtt ctttgtatct gttggtcttc ctaatatgac 
                 1020 
               
               
                 tcaaggattc tgggaaaatt ccatgctaac ggacccagga aatgttcaga aagcagtctg 
                 1080 
               
               
                 ccatcccaca gcttgggacc tggggaaggg cgacttcagg atccttatgt gcacaaaggt 
                 1140 
               
               
                 gacaatggac gacttcctga cagctcatca tgagatgggg catatccagt atgatatggc 
                 1200 
               
               
                 atatgctgca caaccttttc tgctaagaaa tggagctaat gaaggattcc atgaagctgt 
                 1260 
               
               
                 tggggaaatc atgtcacttt ctgcagccac acctaagcat ttaaaatcca ttggtcttct 
                 1320 
               
               
                 gtcacccgat tttcaagaag acaatgaaac agaaataaac ttcctgctca aacaagcact 
                 1380 
               
               
                 cacgattgtt gggactctgc catttactta catgttagag aagtggaggt ggatggtctt 
                 1440 
               
               
                 taaaggggaa attcccaaag accagtggat gaaaaagtgg tgggagatga agcgagagat 
                 1500 
               
               
                 agttggggtg gtggaacctg tgccccatga tgaaacatac tgtgaccccg catctctgtt 
                 1560 
               
               
                 ccatgtttct aatgattact cattcattcg atattacaca aggacccttt accaattcca 
                 1620 
               
               
                 gtttcaagaa gcactttgtc aagcagctaa acatgaaggc cctctgcaca aatgtgacat 
                 1680 
               
               
                 ctcaaactct acagaagctg gacagaaact gttcaatatg ctgaggcttg gaaaatcaga 
                 1740 
               
               
                 accctggacc ctagcattgg aaaatgttgt aggagcaaag aacatgaatg taaggccact 
                 1800 
               
               
                 gctcaactac tttgagccct tatttacctg gctgaaagac cagaacaaga attcttttgt 
                 1860 
               
               
                 gggatggagt accgactgga gtccatatgc agaccaaagc atcaaagtga ggataagcct 
                 1920 
               
               
                 aaaatcagct cttggagata aagcatatga atggaacgac aatgaaatgt acctgttccg 
                 1980 
               
               
                 atcatctgtt gcatatgcta tgaggcagta ctttttaaaa gtaaaaaatc agatgattct 
                 2040 
               
               
                 ttttggggag gaggatgtgc gagtggctaa tttgaaacca agaatctcct ttaatttctt 
                 2100 
               
               
                 tgtcactgca cctaaaaatg tgtctgatat cattcctaga actgaagttg aaaaggccat 
                 2160 
               
               
                 caggatgtcc cggagccgta tcaatgatgc tttccgtctg aatgacaaca gcctagagtt 
                 2220 
               
               
                 tctggggata cagccaacac ttggacctcc taaccagccc cctgtttcca tatggctgat 
                 2280 
               
               
                 tgtttttgga gttgtgatgg gagtgatagt ggttggcatt gtcatcctga tcttcactgg 
                 2340 
               
               
                 gatcagagat cggaagaaga aaaataaagc aagaagtgga gaaaatcctt atgcctccat 
                 2400 
               
               
                 cgatattagc aaaggagaaa ataatccagg attccaaaac actgatgatg ttcagacctc 
                 2460 
               
               
                 cttttagaaa aatctatgtt tttcctcttg aggtgatttt gttgtatgta aatgttaatt 
                 2520 
               
               
                 tcatggtata gaaaatataa gatgataaag atatcattaa atgtcaaaac tatgactctg 
                 2580 
               
               
                 ttcagaaaaa aaattgtcca aagacaacat ggccaaggag agagcatctt cattgacatt 
                 2640 
               
               
                 gctttcagta tttatttctg tctctggatt tgacttctgt tctgtttctt aataaggatt 
                 2700 
               
               
                 ttgtattaga gtatattagg gaaagtgtgt atttggtctc acaggctgtt cagggataat 
                 2760 
               
               
                 ctaaatgtaa atgtctgttg aatttctgaa gttgaaaaca aggatatatc attggagcaa 
                 2820 
               
               
                 gtgttggatc ttgtatggaa tatggatgga tcacttgtaa ggacagtgcc tgggaactgg 
                 2880 
               
               
                 tgtagctgca aggattgaga atggcatgca ttagctcact ttcatttaat ccattgtcaa 
                 2940 
               
               
                 ggatgacatg ctttcttcac agtaactcag ttcaagtact atggtgattt gcctacagtg 
                 3000 
               
               
                 atgtttggaa tcgatcatgc tttcttcaag gtgacaggtc taaagagaga agaatccagg 
                 3060 
               
               
                 gaacaggtag aggacattgc tttttcactt ccaaggtgct tgatcaacat ctccctgaca 
                 3120 
               
               
                 acacaaaact agagccaggg gcctccgtga actcccagag catgcctgat agaaactcat 
                 3180 
               
               
                 ttctactgtt ctctaactgt ggagtgaatg gaaattccaa ctgtatgttc accctctgaa 
                 3240 
               
               
                 gtgggtaccc agtctcttaa atcttttgta tttgctcaca gtgtttgagc agtgctgagc 
                 3300 
               
               
                 acaaagcaga cactcaataa atgctagatt tacacactc 
                 3339 
               
               
                   
               
            
           
           
               
               
            
               
                 &lt;210&gt; 
                  3 
               
               
                 &lt;211&gt; 
                 3566 
               
               
                 &lt;212&gt; 
                 DNA 
               
               
                 &lt;213&gt; 
                 
                   Mus musculus 
                 
               
               
                 &lt;400&gt; 
                  3 
               
            
           
           
               
               
            
               
                 aggcccatga gccctgccat ttaaagtggc tcctctctta cactctggga atgaggacac 
                   60 
               
               
                 ggagccagct gctgaacttc accaggataa ccattaaaat tgctttggag ttcatatttc 
                  120 
               
               
                 cacgatccca tgcctatgga tgccaaggac ttgtcatgga tgcgctttgg atttcataat 
                  180 
               
               
                 gcagagtcat tattacttcc ttgagttctc agctgagttg taagcagtgc ccaacccaag 
                  240 
               
               
                 ttcaaaggct gatgagagag aaaaactcat gaagagattt tactctaggg aaagttgctc 
                  300 
               
               
                 agtggatggg atcttggcgc acggggaaag atgtccagct cctcctggct ccttctcagc 
                  360 
               
               
                 cttgttgctg ttactactgc tcagtccctc accgaggaaa atgccaagac atttttaaac 
                  420 
               
               
                 aactttaatc aggaagctga agacctgtct tatcaaagtt cacttgcttc ttggaattat 
                  480 
               
               
                 aatactaaca ttactgaaga aaatgcccaa aagatgagtg aggctgcagc caaatggtct 
                  540 
               
               
                 gccttttatg aagaacagtc taagactgcc caaagtttct cactacaaga aatccagact 
                  600 
               
               
                 ccgatcatca agcgtcaact acaggccctt cagcaaagtg ggtcttcagc actctcagca 
                  660 
               
               
                 gacaagaaca aacagttgaa cacaattctg aacaccatga gcaccattta cagtactgga 
                  720 
               
               
                 aaagtttgca acccaaagaa cccacaagaa tgcttattac ttgagccagg attggatgaa 
                  780 
               
               
                 ataatggcga caagcacaga ctacaactct aggctctggg catgggaggg ctggagggct 
                  840 
               
               
                 gaggttggca agcagctgag gccgttgtat gaagagtatg tggtcctgaa aaacgagatg 
                  900 
               
               
                 gcaagagcaa acaattataa cgactatggg gattattgga gaggggacta tgaagcagag 
                  960 
               
               
                 ggagcagatg gctacaacta taaccgtaac cagttgattg aagatgtaga acgtaccttc 
                 1020 
               
               
                 gcagagatca agccattgta tgagcatctt catgcctatg tgaggaggaa gttgatggat 
                 1080 
               
               
                 acctaccctt cctacatcag ccccactgga tgcctccctg cccatttgct tggtgatatg 
                 1140 
               
               
                 tggggtagat tttggacaaa tctgtaccct ttgactgttc cctttgcaca gaaaccaaac 
                 1200 
               
               
                 atagatgtta ctgatgcaat gatgaatcag ggctgggatg cagaaaggat atttcaagag 
                 1260 
               
               
                 gcagagaaat tctttgtttc tgttggcctt cctcatatga ctcaaggatt ctgggcaaac 
                 1320 
               
               
                 tctatgctga ctgagccagc agatggccgg aaagttgtct gccaccccac agcttgggat 
                 1380 
               
               
                 ctgggacacg gagacttcag aatcaagatg tgtacaaagg tcacaatgga caacttcttg 
                 1440 
               
               
                 acagcccatc acgagatggg acacatccaa tatgacatgg catatgccag gcaacctttc 
                 1500 
               
               
                 ctgctaagaa acggagccaa tgaagggttc catgaagctg ttggagaaat catgtcactt 
                 1560 
               
               
                 tctgcagcta cccccaagca tctgaaatcc attggtcttc tgccatccga ttttcaagaa 
                 1620 
               
               
                 gatagcgaaa cagagataaa cttcctactg aaacaggcat tgacaattgt tggaacacta 
                 1680 
               
               
                 ccgtttactt acatgttaga gaagtggagg tggatggtct ttcggggtga aattcccaaa 
                 1740 
               
               
                 gagcagtgga tgaaaaagtg gtgggagatg aagcgggaga tcgttggtgt ggtggagcct 
                 1800 
               
               
                 ctgcctcatg atgaaacata ctgtgaccct gcatctctgt tccatgtttc taatgattac 
                 1860 
               
               
                 tcattcattc gatattacac aaggaccatt taccaattcc agtttcaaga agctctttgt 
                 1920 
               
               
                 caagcagcta agtataatgg ttctctgcac aaatgtgaca tctcaaattc cactgaagct 
                 1980 
               
               
                 gggcagaagt tgctcaagat gctgagtctt ggaaattcag agccctggac caaagccttg 
                 2040 
               
               
                 gaaaatgtgg taggagcaag gaatatggat gtaaaaccac tgctcaatta cttccaaccg 
                 2100 
               
               
                 ttgtttgact ggctgaaaga gcagaacaga aattcttttg tggggtggaa cactgaatgg 
                 2160 
               
               
                 agcccatatg ccgaccaaag cattaaagtg aggataagcc taaaatcagc tcttggagct 
                 2220 
               
               
                 aatgcatatg aatggaccaa caacgaaatg ttcctgttcc gatcatctgt tgcatatgcc 
                 2280 
               
               
                 atgagaaagt atttttcaat aatcaaaaac cagacagttc cttttctaga ggaagatgta 
                 2340 
               
               
                 cgagtgagtg atttgaaacc aagagtctcc ttctacttct ttgtcacctc accccaaaat 
                 2400 
               
               
                 gtgtctgatg tcattcctag aagtgaagtt gaagatgcca tcaggatgtc tcggggccgc 
                 2460 
               
               
                 atcaatgatg tctttggcct gaatgataac agcctggagt ttctggggat tcacccaaca 
                 2520 
               
               
                 cttgagccac cttaccagcc tcctgtcacc atatggctga ttatttttgg tgttgtgatg 
                 2580 
               
               
                 gcactggtag tggttggcat catcatcctg attgtcactg ggatcaaagg tcgaaagaag 
                 2640 
               
               
                 aaaaatgaaa caaaaagaga agagaaccct tatgactcga tggacattgg aaaaggagaa 
                 2700 
               
               
                 agcaatgcag gattccaaaa cagtgatgat gctcagactt ccttttagca aagcacttgt 
                 2760 
               
               
                 catcttcctg tatgtaaatg ctaacttcat agtacacaaa atatgagagt atacacatgt 
                 2820 
               
               
                 cattagctat caaaactatg atctgttcag taaacgttgt ccaaagagca tcagacttga 
                 2880 
               
               
                 gtggacatct tcactgacat tgctttcagt atttatttct gcctaaggat ttgacatctc 
                 2940 
               
               
                 ttctgtttat taatagagat gtttatctta gcataaaaga gggaaatgtg cctttggcct 
                 3000 
               
               
                 cacagtctat ccagggtgat atggttgggt aactggagtt agaagatgag atgatgtctc 
                 3060 
               
               
                 ttgggggcaa gtgttggctt cggtgtggca tctgggctgt gaactggtgg gactgttgag 
                 3120 
               
               
                 gttgagaatg gtgctcgctg gtcacttgaa tccaagtgtg acgtcatgct ctgtggcttc 
                 3180 
               
               
                 tgccttcaca cttctcactt caagtactgt aggaatttgt tcacagtaat acttgaaatg 
                 3240 
               
               
                 gactgtccct tctttggagg tgcagttcaa cggagaaaga agccagacat caggtagaga 
                 3300 
               
               
                 ccatgacctt ttctcttcca aacttgatca acatctctct aacaagacac agctagcaca 
                 3360 
               
               
                 ggaaactcca cgaacccaga gcatgcctgt cagaaactac ttccattatt ctcccattgt 
                 3420 
               
               
                 ggagtaaggg aaaattccag atgaatgctc gatctgtgag atgggtgccc agtctctgaa 
                 3480 
               
               
                 attgtttgta ttttctcaca gggtctgagc aatggtgaac acaaagccga cctcaataaa 
                 3540 
               
               
                 tacttattag atttagacac tcctct 
                 3566 
               
               
                   
               
            
           
           
               
               
            
               
                 &lt;210&gt; 
                  4 
               
               
                 &lt;211&gt; 
                 3418 
               
               
                 &lt;212&gt; 
                 DNA 
               
               
                 &lt;213&gt; 
                 
                   Mus musculus 
                 
               
               
                 &lt;400&gt;  
                  4 
               
            
           
           
               
               
            
               
                 ttaacttcat attggtccag cagcttgttt actgttctct tctgtttctt cttctgcttt 
                   60 
               
               
                 ttttttcttc tcttctcagt gcccaaccca agttcaaagg ctgatgagag agaaaaactc 
                  120 
               
               
                 atgaagagat tttactctag ggaaagttgc tcagtggatg ggatcttggc gcacggggaa 
                  180 
               
               
                 agatgtccag ctcctcctgg ctccttctca gccttgttgc tgttactact gctcagtccc 
                  240 
               
               
                 tcaccgagga aaatgccaag acatttttaa acaactttaa tcaggaagct gaagacctgt 
                  300 
               
               
                 cttatcaaag ttcacttgct tcttggaatt ataatactaa cattactgaa gaaaatgccc 
                  360 
               
               
                 aaaagatgag tgaggctgca gccaaatggt ctgcctttta tgaagaacag tctaagactg 
                  420 
               
               
                 cccaaagttt ctcactacaa gaaatccaga ctccgatcat caagcgtcaa ctacaggccc 
                  480 
               
               
                 ttcagcaaag tgggtcttca gcactctcag cagacaagaa caaacagttg aacacaattc 
                  540 
               
               
                 tgaacaccat gagcaccatt tacagtactg gaaaagtttg caacccaaag aacccacaag 
                  600 
               
               
                 aatgcttatt acttgagcca ggattggatg aaataatggc gacaagcaca gactacaact 
                  660 
               
               
                 ctaggctctg ggcatgggag ggctggaggg ctgaggttgg caagcagctg aggccgttgt 
                  720 
               
               
                 atgaagagta tgtggtcctg aaaaacgaga tggcaagagc aaacaattat aacgactatg 
                  780 
               
               
                 gggattattg gagaggggac tatgaagcag agggagcaga tggctacaac tataaccgta 
                  840 
               
               
                 accagttgat tgaagatgta gaacgtacct tcgcagagat caagccattg tatgagcatc 
                  900 
               
               
                 ttcatgccta tgtgaggagg aagttgatgg atacctaccc ttcctacatc agccccactg 
                  960 
               
               
                 gatgcctccc tgcccatttg cttggtgata tgtggggtag attttggaca aatctgtacc 
                 1020 
               
               
                 ctttgactgt tccctttgca cagaaaccaa acatagatgt tactgatgca atgatgaatc 
                 1080 
               
               
                 agggctggga tgcagaaagg atatttcaag aggcagagaa attctttgtt tctgttggcc 
                 1140 
               
               
                 ttcctcatat gactcaagga ttctgggcaa actctatgct gactgagcca gcagatggcc 
                 1200 
               
               
                 ggaaagttgt ctgccacccc acagcttggg atctgggaca cggagacttc agaatcaaga 
                 1260 
               
               
                 tgtgtacaaa ggtcacaatg gacaacttct tgacagccca tcacgagatg ggacacatcc 
                 1320 
               
               
                 aatatgacat ggcatatgcc aggcaacctt tcctgctaag aaacggagcc aatgaagggt 
                 1380 
               
               
                 tccatgaagc tgttggagaa atcatgtcac tttctgcagc tacccccaag catctgaaat 
                 1440 
               
               
                 ccattggtct tctgccatcc gattttcaag aagatagcga aacagagata aacttcctac 
                 1500 
               
               
                 tgaaacaggc attgacaatt gttggaacac taccgtttac ttacatgtta gagaagtgga 
                 1560 
               
               
                 ggtggatggt ctttcggggt gaaattccca aagagcagtg gatgaaaaag tggtgggaga 
                 1620 
               
               
                 tgaagcggga gatcgttggt gtggtggagc ctctgcctca tgatgaaaca tactgtgacc 
                 1680 
               
               
                 ctgcatctct gttccatgtt tctaatgatt actcattcat tcgatattac acaaggacca 
                 1740 
               
               
                 tttaccaatt ccagtttcaa gaagctcttt gtcaagcagc taagtataat ggttctctgc 
                 1800 
               
               
                 acaaatgtga catctcaaat tccactgaag ctgggcagaa gttgctcaag atgctgagtc 
                 1860 
               
               
                 ttggaaattc agagccctgg accaaagcct tggaaaatgt ggtaggagca aggaatatgg 
                 1920 
               
               
                 atgtaaaacc actgctcaat tacttccaac cgttgtttga ctggctgaaa gagcagaaca 
                 1980 
               
               
                 gaaattcttt tgtggggtgg aacactgaat ggagcccata tgccgaccaa agcattaaag 
                 2040 
               
               
                 tgaggataag cctaaaatca gctcttggag ctaatgcata tgaatggacc aacaacgaaa 
                 2100 
               
               
                 tgttcctgtt ccgatcatct gttgcatatg ccatgagaaa gtatttttca ataatcaaaa 
                 2160 
               
               
                 accagacagt tccttttcta gaggaagatg tacgagtgag tgatttgaaa ccaagagtct 
                 2220 
               
               
                 ccttctactt ctttgtcacc tcaccccaaa atgtgtctga tgtcattcct agaagtgaag 
                 2280 
               
               
                 ttgaagatgc catcaggatg tctcggggcc gcatcaatga tgtctttggc ctgaatgata 
                 2340 
               
               
                 acagcctgga gtttctgggg attcacccaa cacttgagcc accttaccag cctcctgtca 
                 2400 
               
               
                 ccatatggct gattattttt ggtgttgtga tggcactggt agtggttggc atcatcatcc 
                 2460 
               
               
                 tgattgtcac tgggatcaaa ggtcgaaaga agaaaaatga aacaaaaaga gaagagaacc 
                 2520 
               
               
                 cttatgactc gatggacatt ggaaaaggag aaagcaatgc aggattccaa aacagtgatg 
                 2580 
               
               
                 atgctcagac ttccttttag caaagcactt gtcatcttcc tgtatgtaaa tgctaacttc 
                 2640 
               
               
                 atagtacaca aaatatgaga gtatacacat gtcattagct atcaaaacta tgatctgttc 
                 2700 
               
               
                 agtaaacgtt gtccaaagag catcagactt gagtggacat cttcactgac attgctttca 
                 2760 
               
               
                 gtatttattt ctgcctaagg atttgacatc tcttctgttt attaatagag atgtttatct 
                 2820 
               
               
                 tagcataaaa gagggaaatg tgcctttggc ctcacagtct atccagggtg atatggttgg 
                 2880 
               
               
                 gtaactggag ttagaagatg agatgatgtc tcttgggggc aagtgttggc ttcggtgtgg 
                 2940 
               
               
                 catctgggct gtgaactggt gggactgttg aggttgagaa tggtgctcgc tggtcacttg 
                 3000 
               
               
                 aatccaagtg tgacgtcatg ctctgtggct tctgccttca cacttctcac ttcaagtact 
                 3060 
               
               
                 gtaggaattt gttcacagta atacttgaaa tggactgtcc cttctttgga ggtgcagttc 
                 3120 
               
               
                 aacggagaaa gaagccagac atcaggtaga gaccatgacc ttttctcttc caaacttgat 
                 3180 
               
               
                 caacatctct ctaacaagac acagctagca caggaaactc cacgaaccca gagcatgcct 
                 3240 
               
               
                 gtcagaaact acttccatta ttctcccatt gtggagtaag ggaaaattcc agatgaatgc 
                 3300 
               
               
                 tcgatctgtg agatgggtgc ccagtctctg aaattgtttg tattttctca cagggtctga 
                 3360 
               
               
                 gcaatggtga acacaaagcc gacctcaata aatacttatt agatttagac actcctct 
                 3418 
               
               
                   
               
            
           
           
               
               
            
               
                 &lt;210&gt; 
                  5 
               
               
                 &lt;211&gt; 
                 2418 
               
               
                 &lt;212&gt; 
                 DNA 
               
               
                 &lt;213&gt; 
                 
                   Rattus norvegicus 
                 
               
               
                 &lt;400&gt; 
                  5 
               
            
           
           
               
               
            
               
                 atgtcaagct cctgctggct ccttctcagc cttgttgctg ttgctactgc tcagtccctc 
                   60 
               
               
                 atcgaggaaa aggccgagag ctttttaaac aagtttaacc aggaagctga agacctgtct 
                  120 
               
               
                 tatcaaagtt cacttgcttc ttggaattac aacaccaaca ttacggagga gaatgcccaa 
                  180 
               
               
                 aagatgaacg aggctgcggc caaatggtct gccttttatg aagaacagtc caagatcgcc 
                  240 
               
               
                 caaaatttct cactacaaga aattcagaat gcgaccatca agcgtcaact gaaggccctt 
                  300 
               
               
                 cagcagagcg ggtcttcagc gctgtcacca gacaagaaca aacagttgaa cacaattcta 
                  360 
               
               
                 aacaccatga gcaccattta cagtactgga aaagtttgca actcaatgaa tccacaagaa 
                  420 
               
               
                 tgttttttac ttgaaccagg attggacgaa ataatggcaa caagcacaga ctacaatcgt 
                  480 
               
               
                 aggctctggg cttgggaggg ctggagggct gaggtcggca agcagctgag gccgttatat 
                  540 
               
               
                 gaagagtatg tggtcctgaa aaatgagatg gcaagagcaa acaattatga agactatggg 
                  600 
               
               
                 gattattggc gaggggatta tgaagcagag ggagtagaag gttacaacta caaccgaaac 
                  660 
               
               
                 cagttgatcg aagacgtaga aaataccttc aaagagatca aaccgttgta tgagcaactt 
                  720 
               
               
                 catgcctatg tgagaacgaa gttgatggaa gtgtaccctt cttacatcag ccctactgga 
                  780 
               
               
                 tgcctccctg ctcatttgct tggtgatatg tggggtaggt tttggacaaa tctgtaccct 
                  840 
               
               
                 ttgactactc cctttcttca gaaaccaaac atagatgtta ctgatgcaat ggtgaatcag 
                  900 
               
               
                 agctgggatg cagaaagaat atttaaagag gcagagaagt tcttcgtttc tgttggcctt 
                  960 
               
               
                 cctcaaatga ctccgggatt ctggacaaac tccatgctga ctgagccagg agatgaccgg 
                 1020 
               
               
                 aaagttgtct gccaccccac agcttgggat ctgggacatg gagacttcag aatcaagatg 
                 1080 
               
               
                 tgcacaaagg tcacaatgga caacttcttg acagcccatc atgagatggg acacatccaa 
                 1140 
               
               
                 tatgacatgg catatgccaa gcaacctttc ctgctaagaa acggagccaa tgaagggttc 
                 1200 
               
               
                 catgaagccg ttggagaaat catgtcactt tctgcagcta cccccaaaca tttgaaatct 
                 1260 
               
               
                 attggtcttc tgccatccaa ttttcaagaa gacaatgaaa cagaaataaa cttcctactc 
                 1320 
               
               
                 aaacaggcat tgacaattgt tggaacgctg ccatttactt acatgttaga gaagtggagg 
                 1380 
               
               
                 tggatggtct ttcaggataa aattcccaga gaacagtgga ccaaaaagtg gtgggagatg 
                 1440 
               
               
                 aagcgggaga tcgttggtgt ggtggagcct ctgcctcatg atgaaacata ctgtgaccct 
                 1500 
               
               
                 gcatctctgt tccatgtctc taatgattac tcattcattc gatattacac aaggaccatt 
                 1560 
               
               
                 tatcaattcc agtttcaaga agctctttgt caagcagcta aacatgatgg cccactacac 
                 1620 
               
               
                 aaatgtgaca tctcaaattc cactgaagct gggcagaagt tgctcaatat gctgagtctt 
                 1680 
               
               
                 ggaaactcag ggccctggac cctagccttg gaaaatgtgg taggatcaag gaatatggat 
                 1740 
               
               
                 gtaaaaccac tgctcaatta cttccaacca ttgtttgtct ggctgaaaga gcagaacagg 
                 1800 
               
               
                 aattcgactg tggggtggag cactgactgg agcccatatg ccgaccaaag cattaaagtg 
                 1860 
               
               
                 aggataagcc taaaatcagc tcttgggaaa aatgcgtatg aatggaccga caacgaaatg 
                 1920 
               
               
                 tacctattcc gatcatctgt tgcctatgcc atgagagagt atttttcaag ggaaaagaac 
                 1980 
               
               
                 cagacagttc cttttgggga ggcagacgta tgggtgagtg atttgaaacc aagagtctcc 
                 2040 
               
               
                 ttcaacttct ttgtcacttc acccaaaaat gtgtctgaca tcattcccag aagtgaagtt 
                 2100 
               
               
                 gaagaggcca tcaggatgtc tcggggccgt atcaatgata tttttggtct gaatgataac 
                 2160 
               
               
                 agcctggagt ttctggggat ctacccaaca cttaagccac cttacgagcc tcctgtcacc 
                 2220 
               
               
                 atatggctga ttatttttgg tgtcgtgatg ggaacggtag tggttggcat tgttatcctg 
                 2280 
               
               
                 atcgtcactg ggatcaaagg tcgaaagaag aaaaatgaaa caaaaagaga agagaatcct 
                 2340 
               
               
                 tatgactcca tggacattgg caaaggagaa agtaacgcag gattccaaaa cagtgatgat 
                 2400 
               
               
                 gctcaaactt cattctaa 
                 2418 
               
               
                   
               
            
           
           
               
               
            
               
                 &lt;210&gt;  
                  6 
               
               
                 &lt;211&gt; 
                 3589 
               
               
                 &lt;212&gt; 
                 DNA 
               
               
                 &lt;213&gt; 
                 
                   Macaca fascicularis 
                 
               
               
                 &lt;400&gt;  
                  6 
               
            
           
           
               
               
            
               
                 catacataca ctctagtaat gaggacactg agctcgcgtc tgaaatttaa caagataacc 
                   60 
               
               
                 actaaaatct ctttgaattc tatattgttg tgatcctgtg gctacagaga atcaggagtt 
                  120 
               
               
                 gacgtagaca ctctttggat ttcatactgt atggaggctt tcttacttcc acgtgacctt 
                  180 
               
               
                 gactgagttt tgaatagtgc ccaacccaag ttcaaaggct gataagagag aaaatctcat 
                  240 
               
               
                 gaggaggttt tagtctaggg aaagtcattc agtggatgtg atcttggctc acaggggacg 
                  300 
               
               
                 atgtcaggct cttcctggct ccttctcagc cttgttgctg taactgctgc tcagtccacc 
                  360 
               
               
                 attgaggaac aggccaagac atttttggac aagtttaacc acgaagccga agacctgttc 
                  420 
               
               
                 tatcaaagtt cacttgcttc ttggaattat aacaccaata ttactgaaga gaatgtccaa 
                  480 
               
               
                 aacatgaata atgctgggga aaaatggtct gcctttttaa aagaacagtc cacacttgcc 
                  540 
               
               
                 caaatgtatc cactgcaaga aattcagaat ctcacagtca agcttcagtt gcaggctctt 
                  600 
               
               
                 cagcaaaatg ggtcttcagt gctctcagaa gacaagagca aacggttgaa cacaattcta 
                  660 
               
               
                 aatacaatga gcaccatcta cagtactgga aaagtttgta acccaaataa tccccaggaa 
                  720 
               
               
                 tgcttattac ttgatccagg tttgaatgaa ataatggaaa agagtttaga ctacaatgag 
                  780 
               
               
                 aggctctggg cttgggaagg ctggagatct gaggtcggca agcagctgag gccattatat 
                  840 
               
               
                 gaagagtatg tggtcttgaa aaatgagatg gcaagagcaa atcattataa ggactatggg 
                  900 
               
               
                 gattattgga gaggaaacta tgaagtaaac ggggtagatg gctatgacta caaccgcgac 
                  960 
               
               
                 cagttgattg aagatgtgga acgtaccttt gaagagatta aaccattata tgaacatctt 
                 1020 
               
               
                 catgcctatg tgagggcaaa gttgatgaat gcctaccctt cctatattag tccaactgga 
                 1080 
               
               
                 tgccttcctg ctcatttgct tggtgatatg tggggtagat tttggacaaa tctgtactct 
                 1140 
               
               
                 ttgacagttc cctttggaca gaaaccaaac atagatgtta ctgatgcaat ggtgaaccag 
                 1200 
               
               
                 gcctggaatg cacagagaat attcaaggag gccgagaagt tctttgtatc tgttggtctt 
                 1260 
               
               
                 cctaatatga ctcaaggatt ctgggaaaat tccatgctaa ctgatccagg aaatgttcag 
                 1320 
               
               
                 aaagtagtct gccaccccac agcttgggac ctggggaagg gtgacttcag gatcattatg 
                 1380 
               
               
                 tgcacaaagg tgacaatgga cgacttcctg acagctcatc atgagatggg gcatatccaa 
                 1440 
               
               
                 tatgatatgg catatgctgc acaacctttt ctgctaagaa atggagctaa tgaaggattc 
                 1500 
               
               
                 catgaagctg ttggggaaat catgtcactt tctgcagcca cacctaagca tttaaaatcc 
                 1560 
               
               
                 attggtcttc tgtcacctga ttttcaagaa gacaatgaaa cagaaataaa cttcctgctc 
                 1620 
               
               
                 aaacaagcac tcacgattgt tgggactctg ccatttactt acatgttaga gaagtggagg 
                 1680 
               
               
                 tggatggtct ttaaaggtga aattcccaaa gaccagtgga tgaaaaagtg gtgggagatg 
                 1740 
               
               
                 aagcgagaga tagttggggt ggtggaacct gtgccccatg atgaaacata ctgtgacccc 
                 1800 
               
               
                 gcatctctgt tccatgtttc taatgattac tcattcattc gatattacac aaggaccctt 
                 1860 
               
               
                 taccaattcc agtttcaaga agcactttgt caagcagcta aacacgaagg ccctctgcac 
                 1920 
               
               
                 aaatgtgaca tctcaaactc tacagaagct ggacagaaat tgctcaatat gctgaagctt 
                 1980 
               
               
                 ggaaaatcag aaccctggac cctagcattg gaaaatgttg taggagcaaa gaacatgaat 
                 2040 
               
               
                 gtaagaccac tgctcaacta ctttgagccc ttgtttacct ggctgaaaga ccagaacaag 
                 2100 
               
               
                 aattcttttg tgggatggag taccgactgg agtccgtatg ctgaccaaag catcaaagtg 
                 2160 
               
               
                 aggataagct taaaatcagc tcttggagat aaagcatatg aatggaacga caatgaaatg 
                 2220 
               
               
                 tacctgttcc gatcatctgt tgcatatgcc atgaggacgt actttttaga aatcaaacat 
                 2280 
               
               
                 cagacgattc tttttgggga ggaggatgtg cgagttgctg atttgaaacc aagaatctcc 
                 2340 
               
               
                 tttaatttct atgtcactgc acctaaaaat gtgtctgaca tcattcctag aactgaagtt 
                 2400 
               
               
                 gaagaggcca tcaggatctc caggagccgt atcaatgatg ctttccgtct gaatgacaac 
                 2460 
               
               
                 agcctggagt ttctggggat acagacaaca cttgcacctc cttaccagtc ccccgttacc 
                 2520 
               
               
                 acatggctaa ttgtttttgg agttgtgatg ggagtgatag tggctggcat tgtcgtcctg 
                 2580 
               
               
                 atcttcactg ggatcagaga tcgaaagaag aaaaatcaag caagaagtga agaaaatcct 
                 2640 
               
               
                 tatgcctcca tcgatattaa caaaggagaa aataatccag gattccaaaa cactgatgat 
                 2700 
               
               
                 gttcagacct ccttttagaa aaatctatgt tttttctctt gaggtgattt tgttgtaggt 
                 2760 
               
               
                 aaatgttaat ttcatggtat cgaacatatg agatgataaa tatatcatta aatgtcaaaa 
                 2820 
               
               
                 ctatgaatct gttcagaaaa aaattgtcca aagacaacat ggtcaaggag agagcatctt 
                 2880 
               
               
                 cattgacatt gctttcagta tttatttctg tctctggatt tgacttctgt tctgtttcct 
                 2940 
               
               
                 aataaggatt ttgtattaga gtgtattagg gaaagtgtgt atttggtctc acgggctgtt 
                 3000 
               
               
                 cagggataat ctaaatgtaa atgtctgttg aatttctgaa gttgaaaaca aggatatatc 
                 3060 
               
               
                 atgggagcaa gtattggacc ttgtatggaa tatggatgga tcacttgtaa ggacagtgcc 
                 3120 
               
               
                 tgggaactgg tacagctgca aggattgaga atggcatgca ctagttcact ttcatttaat 
                 3180 
               
               
                 ccattgttaa ggatgacata ctttcttcac attaacttaa tccaagtact atggtgattt 
                 3240 
               
               
                 gcctacagtg atgtttggaa tagatcacgc tttcttcaag gtgacaggtc taaagagaga 
                 3300 
               
               
                 agaatccagg gaacaggtag aggacattgc tttttcactt ccaagctgct tgatcaacat 
                 3360 
               
               
                 ctccctgata acacaaaact aacgccaggg gcctccgtga actcccagag catgcctgat 
                 3420 
               
               
                 agaaactcat ttccactgtt ctctaactgt ggagtgaatg gaaattccaa ctgtatgttc 
                 3480 
               
               
                 accctctgaa gtgggtatcc agtctcttaa atcttttgta tttgcccaca gtgtttgagt 
                 3540 
               
               
                 agtgctgagc acaaagcaga cactcaataa atgctagatt tgcacactc 
                 3589 
               
               
                   
               
            
           
           
               
               
            
               
                 &lt;210&gt; 
                  7 
               
               
                 &lt;211&gt; 
                 3596 
               
               
                 &lt;212&gt; 
                 DNA 
               
               
                 &lt;213&gt; 
                 
                   Homo sapiens 
                 
               
               
                 &lt;400&gt; 
                  7 
               
            
           
           
               
               
            
               
                 gagtgtgtaa atctagcatt tattgagtgt ctgctttgtg ctcagcactg ctcaaacact 
                   60 
               
               
                 gtgagcaaat acaaaagatt taagagactg ggtacccact tcagagggtg aacatacagt 
                  120 
               
               
                 tggaatttcc attcactcca cagttagaga acagtagaaa tgagtttcta tcaggcatgc 
                  180 
               
               
                 tctgggagtt cacggaggcc cctggctcta gttttgtgtt gtcagggaga tgttgatcaa 
                  240 
               
               
                 gcaccttgga agtgaaaaag caatgtcctc tacctgttcc ctggattctt ctctctttag 
                  300 
               
               
                 acctgtcacc ttgaagaaag catgatcgat tccaaacatc actgtaggca aatcaccata 
                  360 
               
               
                 gtacttgaac tgagttactg tgaagaaagc atgtcatcct tgacaatgga ttaaatgaaa 
                  420 
               
               
                 gtgagctaat gcatgccatt ctcaatcctt gcagctacac cagttcccag gcactgtcct 
                  480 
               
               
                 tacaagtgat ccatccatat tccatacaag atccaacact tgctccaatg atatatcctt 
                  540 
               
               
                 gttttcaact tcagaaattc aacagacatt tacatttaga ttatccctga acagcctgtg 
                  600 
               
               
                 agaccaaata cacactttcc ctaatatact ctaatacaaa atccttatta agaaacagaa 
                  660 
               
               
                 cagaagtcaa atccagagac agaaataaat actgaaagca atgtcaatga agatgctctc 
                  720 
               
               
                 tccttggcca tgttgtcttt ggacaatttt ttttctgaac agagtcatag ttttgacatt 
                  780 
               
               
                 taatgatatc tttatcatct tatattttct ataccatgaa attaacattt acatacaaca 
                  840 
               
               
                 aaatcacctc aagaggaaaa acatagattt ttctaaaagg aggtctgaac atcatcagtg 
                  900 
               
               
                 ttttggaatc ctggattatt ttctcctttg ctaatatcga tggaggcata aggattttct 
                  960 
               
               
                 ccacttcttg ctttattttt cttcttccga tctctgatcc cagtgaagat caggatgaca 
                 1020 
               
               
                 atgccaacca ctatcactcc catcacaact ccaaaaacaa tcagccatat ggaaacaggg 
                 1080 
               
               
                 ggctggttag gaggtccaag tgttggctgt atccccagaa actctaggct gttgtcattc 
                 1140 
               
               
                 agacggaaag catcattgat acggctccgg gacatcctga tggccttttc aacttcagtt 
                 1200 
               
               
                 ctaggaatga tatcagacac atttttaggt gcagtgacaa agaaattaaa ggagattctt 
                 1260 
               
               
                 ggtttcaaat tagccactcg cacatcctcc tccccaaaaa gaatcatctg attttttact 
                 1320 
               
               
                 tttaaaaagt actgcctcat agcatatgca acagatgatc ggaacaggta catttcattg 
                 1380 
               
               
                 tcgttccatt catatgcttt atctccaaga gctgatttta ggcttatcct cactttgatg 
                 1440 
               
               
                 ctttggtctg catatggact ccagtcggta ctccatccca caaaagaatt cttgttctgg 
                 1500 
               
               
                 tctttcagcc aggtaaataa gggctcaaag tagttgagca gtggccttac attcatgttc 
                 1560 
               
               
                 tttgctccta caacattttc caatgctagg gtccagggtt ctgattttcc aagcctcagc 
                 1620 
               
               
                 atattgaaca gtttctgtcc agcttctgta gagtttgaga tgtcacattt gtgcagaggg 
                 1680 
               
               
                 ccttcatgtt tagctgcttg acaaagtgct tcttgaaact ggaattggta aagggtcctt 
                 1740 
               
               
                 gtgtaatatc gaatgaatga gtaatcatta gaaacatgga acagagatgc ggggtcacag 
                 1800 
               
               
                 tatgtttcat catggggcac aggttccacc accccaacta tctctcgctt catctcccac 
                 1860 
               
               
                 cactttttca tccactggtc tttgggaatt tcccctttaa agaccatcca cctccacttc 
                 1920 
               
               
                 tctaacatgt aagtaaatgg cagagtccca acaatcgtga gtgcttgttt gagcaggaag 
                 1980 
               
               
                 tttatttctg tttcattgtc ttcttgaaaa tcgggtgaca gaagaccaat ggattttaaa 
                 2040 
               
               
                 tgcttaggtg tggctgcaga aagtgacatg atttccccaa cagcttcatg gaatccttca 
                 2100 
               
               
                 ttagctccat ttcttagcag aaaaggttgt gcagcatatg ccatatcata ctggatatgc 
                 2160 
               
               
                 cccatctcat gatgagctgt caggaagtcg tccattgtca cctttgtgca cataaggatc 
                 2220 
               
               
                 ctgaagtcgc ccttccccag gtcccaagct gtgggatggc agactgcttt ctgaacattt 
                 2280 
               
               
                 cctgggtccg ttagcatgga attttcccag aatccttgag tcatattagg aagaccaaca 
                 2340 
               
               
                 gatacaaaga acttctcggc ctccttgaat attctctgtg catcccaggc ctggtccacc 
                 2400 
               
               
                 attgcatcag taacatctat gtttggtttc tgtccaaagg gaactgtcaa agagtacaga 
                 2460 
               
               
                 tttgtccaaa atctacccca catatcacca agcaaatgag cagggaggca tccaattgga 
                 2520 
               
               
                 ctgatatagg aaggataggc attcatcaac tttgccctca cataggcatg aagatgttca 
                 2580 
               
               
                 tataatggtt taatctcttc aaaggtatgt tccacatctt caatcaactg gccgcggctg 
                 2640 
               
               
                 tagtcatagc catctacccc atttacttca tagtctcctc tccaataatc cccatagtcc 
                 2700 
               
               
                 tcataatgat ttgctcttgc catctcattt ttcaagacca catactcttc atataatggc 
                 2760 
               
               
                 ctcagctgct tgccgacctc agatctccag ctttcccaag cccagagcct ctcattgtag 
                 2820 
               
               
                 tctaaactgt ttgccattat ttcattcaaa cctggttcaa gtaataagca ttcttgtgga 
                 2880 
               
               
                 ttatctgggt tacaaacttt tccagtactg tagatggtgc tcattgtatt tagaattgtg 
                 2940 
               
               
                 ttcaaccgtt tgctcttgtc ttctgagagc actgaagacc cattttgctg aagagcctgc 
                 3000 
               
               
                 agctgaagct tgactgtgag attctgaatt tcttgtagtg gatacatttg ggcaagtgtg 
                 3060 
               
               
                 gactgttcct ttaaaaaggc agaccatttg tccccagcat tattcatgtt ttggacattc 
                 3120 
               
               
                 tcttcagtaa tattggtgtt ataattccaa gaagcaagtg aactttgata gaacaggtct 
                 3180 
               
               
                 tcggcttcgt ggttaaactt gtccaaaaat gtcttggcct gttcctcaat ggtggactga 
                 3240 
               
               
                 gcagcagtta cagcaacaag gctgagaagg agccaggaag agcttgacat cgtcccctgt 
                 3300 
               
               
                 gagccaagat cacatccact gaatgacttt ccctagacta aaacctcctc atgagatttt 
                 3360 
               
               
                 ctctcttatc agcctttgaa cttgggttgg gcgctattca aaactcagtc aaggtcacgt 
                 3420 
               
               
                 ggaagtaaga aagcctccac atggtatgaa atccaaagag tatctatgtc aactcctgat 
                 3480 
               
               
                 cctctgtagc catgggatca caacaacata gaattcaaag agattttagt ggttatcttg 
                 3540 
               
               
                 tcaaatttca gaagcgagct cagtgtcctc attgccagag tgtatgtatg agtgcc 
                 3596 
               
               
                   
               
            
           
           
               
               
            
               
                 &lt;210&gt; 
                  8 
               
               
                 &lt;211&gt; 
                 3339 
               
               
                 &lt;212&gt; 
                 DNA 
               
               
                 &lt;213&gt; 
                 
                   Homo sapiens 
                 
               
               
                 &lt;400&gt; 
                  8 
               
            
           
           
               
               
            
               
                 gagtgtgtaa atctagcatt tattgagtgt ctgctttgtg ctcagcactg ctcaaacact 
                   60 
               
               
                 gtgagcaaat acaaaagatt taagagactg ggtacccact tcagagggtg aacatacagt 
                  120 
               
               
                 tggaatttcc attcactcca cagttagaga acagtagaaa tgagtttcta tcaggcatgc 
                  180 
               
               
                 tctgggagtt cacggaggcc cctggctcta gttttgtgtt gtcagggaga tgttgatcaa 
                  240 
               
               
                 gcaccttgga agtgaaaaag caatgtcctc tacctgttcc ctggattctt ctctctttag 
                  300 
               
               
                 acctgtcacc ttgaagaaag catgatcgat tccaaacatc actgtaggca aatcaccata 
                  360 
               
               
                 gtacttgaac tgagttactg tgaagaaagc atgtcatcct tgacaatgga ttaaatgaaa 
                  420 
               
               
                 gtgagctaat gcatgccatt ctcaatcctt gcagctacac cagttcccag gcactgtcct 
                  480 
               
               
                 tacaagtgat ccatccatat tccatacaag atccaacact tgctccaatg atatatcctt 
                  540 
               
               
                 gttttcaact tcagaaattc aacagacatt tacatttaga ttatccctga acagcctgtg 
                  600 
               
               
                 agaccaaata cacactttcc ctaatatact ctaatacaaa atccttatta agaaacagaa 
                  660 
               
               
                 cagaagtcaa atccagagac agaaataaat actgaaagca atgtcaatga agatgctctc 
                  720 
               
               
                 tccttggcca tgttgtcttt ggacaatttt ttttctgaac agagtcatag ttttgacatt 
                  780 
               
               
                 taatgatatc tttatcatct tatattttct ataccatgaa attaacattt acatacaaca 
                  840 
               
               
                 aaatcacctc aagaggaaaa acatagattt ttctaaaagg aggtctgaac atcatcagtg 
                  900 
               
               
                 ttttggaatc ctggattatt ttctcctttg ctaatatcga tggaggcata aggattttct 
                  960 
               
               
                 ccacttcttg ctttattttt cttcttccga tctctgatcc cagtgaagat caggatgaca 
                 1020 
               
               
                 atgccaacca ctatcactcc catcacaact ccaaaaacaa tcagccatat ggaaacaggg 
                 1080 
               
               
                 ggctggttag gaggtccaag tgttggctgt atccccagaa actctaggct gttgtcattc 
                 1140 
               
               
                 agacggaaag catcattgat acggctccgg gacatcctga tggccttttc aacttcagtt 
                 1200 
               
               
                 ctaggaatga tatcagacac atttttaggt gcagtgacaa agaaattaaa ggagattctt 
                 1260 
               
               
                 ggtttcaaat tagccactcg cacatcctcc tccccaaaaa gaatcatctg attttttact 
                 1320 
               
               
                 tttaaaaagt actgcctcat agcatatgca acagatgatc ggaacaggta catttcattg 
                 1380 
               
               
                 tcgttccatt catatgcttt atctccaaga gctgatttta ggcttatcct cactttgatg 
                 1440 
               
               
                 ctttggtctg catatggact ccagtcggta ctccatccca caaaagaatt cttgttctgg 
                 1500 
               
               
                 tctttcagcc aggtaaataa gggctcaaag tagttgagca gtggccttac attcatgttc 
                 1560 
               
               
                 tttgctccta caacattttc caatgctagg gtccagggtt ctgattttcc aagcctcagc 
                 1620 
               
               
                 atattgaaca gtttctgtcc agcttctgta gagtttgaga tgtcacattt gtgcagaggg 
                 1680 
               
               
                 ccttcatgtt tagctgcttg acaaagtgct tcttgaaact ggaattggta aagggtcctt 
                 1740 
               
               
                 gtgtaatatc gaatgaatga gtaatcatta gaaacatgga acagagatgc ggggtcacag 
                 1800 
               
               
                 tatgtttcat catggggcac aggttccacc accccaacta tctctcgctt catctcccac 
                 1860 
               
               
                 cactttttca tccactggtc tttgggaatt tcccctttaa agaccatcca cctccacttc 
                 1920 
               
               
                 tctaacatgt aagtaaatgg cagagtccca acaatcgtga gtgcttgttt gagcaggaag 
                 1980 
               
               
                 tttatttctg tttcattgtc ttcttgaaaa tcgggtgaca gaagaccaat ggattttaaa 
                 2040 
               
               
                 tgcttaggtg tggctgcaga aagtgacatg atttccccaa cagcttcatg gaatccttca 
                 2100 
               
               
                 ttagctccat ttcttagcag aaaaggttgt gcagcatatg ccatatcata ctggatatgc 
                 2160 
               
               
                 cccatctcat gatgagctgt caggaagtcg tccattgtca cctttgtgca cataaggatc 
                 2220 
               
               
                 ctgaagtcgc ccttccccag gtcccaagct gtgggatggc agactgcttt ctgaacattt 
                 2280 
               
               
                 cctgggtccg ttagcatgga attttcccag aatccttgag tcatattagg aagaccaaca 
                 2340 
               
               
                 gatacaaaga acttctcggc ctccttgaat attctctgtg catcccaggc ctggtccacc 
                 2400 
               
               
                 attgcatcag taacatctat gtttggtttc tgtccaaagg gaactgtcaa agagtacaga 
                 2460 
               
               
                 tttgtccaaa atctacccca catatcacca agcaaatgag cagggaggca tccaattgga 
                 2520 
               
               
                 ctgatatagg aaggataggc attcatcaac tttgccctca cataggcatg aagatgttca 
                 2580 
               
               
                 tataatggtt taatctcttc aaaggtatgt tccacatctt caatcaactg gccgcggctg 
                 2640 
               
               
                 tagtcatagc catctacccc atttacttca tagtctcctc tccaataatc cccatagtcc 
                 2700 
               
               
                 tcataatgat ttgctcttgc catctcattt ttcaagacca catactcttc atataatggc 
                 2760 
               
               
                 ctcagctgct tgccgacctc agatctccag ctttcccaag cccagagcct ctcattgtag 
                 2820 
               
               
                 tctaaactgt ttgccattat ttcattcaaa cctggttcaa gtaataagca ttcttgtgga 
                 2880 
               
               
                 ttatctgggt tacaaacttt tccagtactg tagatggtgc tcattgtatt tagaattgtg 
                 2940 
               
               
                 ttcaaccgtt tgctcttgtc ttctgagagc actgaagacc cattttgctg aagagcctgc 
                 3000 
               
               
                 agctgaagct tgactgtgag attctgaatt tcttgtagtg gatacatttg ggcaagtgtg 
                 3060 
               
               
                 gactgttcct ttaaaaaggc agaccatttg tccccagcat tattcatgtt ttggacattc 
                 3120 
               
               
                 tcttcagtaa tattggtgtt ataattccaa gaagcaagtg aactttgata gaacaggtct 
                 3180 
               
               
                 tcggcttcgt ggttaaactt gtccaaaaat gtcttggcct gttcctcaat ggtggactga 
                 3240 
               
               
                 gcagcagtta cagcaacaag gctgagaagg agccaggaag agcttgacat cgtcccctgt 
                 3300 
               
               
                 gagccaagat cacatccact gaatgacttt ccctagact 
                 3339 
               
               
                   
               
            
           
           
               
               
            
               
                 &lt;210&gt; 
                  9 
               
               
                 &lt;211&gt; 
                 3566 
               
               
                 &lt;212&gt; 
                 DNA 
               
               
                 &lt;213&gt; 
                 
                   Mus musculus 
                 
               
               
                 &lt;400&gt; 
                  9 
               
            
           
           
               
               
            
               
                 agaggagtgt ctaaatctaa taagtattta ttgaggtcgg ctttgtgttc accattgctc 
                   60 
               
               
                 agaccctgtg agaaaataca aacaatttca gagactgggc acccatctca cagatcgagc 
                  120 
               
               
                 attcatctgg aattttccct tactccacaa tgggagaata atggaagtag tttctgacag 
                  180 
               
               
                 gcatgctctg ggttcgtgga gtttcctgtg ctagctgtgt cttgttagag agatgttgat 
                  240 
               
               
                 caagtttgga agagaaaagg tcatggtctc tacctgatgt ctggcttctt tctccgttga 
                  300 
               
               
                 actgcacctc caaagaaggg acagtccatt tcaagtatta ctgtgaacaa attcctacag 
                  360 
               
               
                 tacttgaagt gagaagtgtg aaggcagaag ccacagagca tgacgtcaca cttggattca 
                  420 
               
               
                 agtgaccagc gagcaccatt ctcaacctca acagtcccac cagttcacag cccagatgcc 
                  480 
               
               
                 acaccgaagc caacacttgc ccccaagaga catcatctca tcttctaact ccagttaccc 
                  540 
               
               
                 aaccatatca ccctggatag actgtgaggc caaaggcaca tttccctctt ttatgctaag 
                  600 
               
               
                 ataaacatct ctattaataa acagaagaga tgtcaaatcc ttaggcagaa ataaatactg 
                  660 
               
               
                 aaagcaatgt cagtgaagat gtccactcaa gtctgatgct ctttggacaa cgtttactga 
                  720 
               
               
                 acagatcata gttttgatag ctaatgacat gtgtatactc tcatattttg tgtactatga 
                  780 
               
               
                 agttagcatt tacatacagg aagatgacaa gtgctttgct aaaaggaagt ctgagcatca 
                  840 
               
               
                 tcactgtttt ggaatcctgc attgctttct ccttttccaa tgtccatcga gtcataaggg 
                  900 
               
               
                 ttctcttctc tttttgtttc atttttcttc tttcgacctt tgatcccagt gacaatcagg 
                  960 
               
               
                 atgatgatgc caaccactac cagtgccatc acaacaccaa aaataatcag ccatatggtg 
                 1020 
               
               
                 acaggaggct ggtaaggtgg ctcaagtgtt gggtgaatcc ccagaaactc caggctgtta 
                 1080 
               
               
                 tcattcaggc caaagacatc attgatgcgg ccccgagaca tcctgatggc atcttcaact 
                 1140 
               
               
                 tcacttctag gaatgacatc agacacattt tggggtgagg tgacaaagaa gtagaaggag 
                 1200 
               
               
                 actcttggtt tcaaatcact cactcgtaca tcttcctcta gaaaaggaac tgtctggttt 
                 1260 
               
               
                 ttgattattg aaaaatactt tctcatggca tatgcaacag atgatcggaa caggaacatt 
                 1320 
               
               
                 tcgttgttgg tccattcata tgcattagct ccaagagctg attttaggct tatcctcact 
                 1380 
               
               
                 ttaatgcttt ggtcggcata tgggctccat tcagtgttcc accccacaaa agaatttctg 
                 1440 
               
               
                 ttctgctctt tcagccagtc aaacaacggt tggaagtaat tgagcagtgg ttttacatcc 
                 1500 
               
               
                 atattccttg ctcctaccac attttccaag gctttggtcc agggctctga atttccaaga 
                 1560 
               
               
                 ctcagcatct tgagcaactt ctgcccagct tcagtggaat ttgagatgtc acatttgtgc 
                 1620 
               
               
                 agagaaccat tatacttagc tgcttgacaa agagcttctt gaaactggaa ttggtaaatg 
                 1680 
               
               
                 gtccttgtgt aatatcgaat gaatgagtaa tcattagaaa catggaacag agatgcaggg 
                 1740 
               
               
                 tcacagtatg tttcatcatg aggcagaggc tccaccacac caacgatctc ccgcttcatc 
                 1800 
               
               
                 tcccaccact ttttcatcca ctgctctttg ggaatttcac cccgaaagac catccacctc 
                 1860 
               
               
                 cacttctcta acatgtaagt aaacggtagt gttccaacaa ttgtcaatgc ctgtttcagt 
                 1920 
               
               
                 aggaagttta tctctgtttc gctatcttct tgaaaatcgg atggcagaag accaatggat 
                 1980 
               
               
                 ttcagatgct tgggggtagc tgcagaaagt gacatgattt ctccaacagc ttcatggaac 
                 2040 
               
               
                 ccttcattgg ctccgtttct tagcaggaaa ggttgcctgg catatgccat gtcatattgg 
                 2100 
               
               
                 atgtgtccca tctcgtgatg ggctgtcaag aagttgtcca ttgtgacctt tgtacacatc 
                 2160 
               
               
                 ttgattctga agtctccgtg tcccagatcc caagctgtgg ggtggcagac aactttccgg 
                 2220 
               
               
                 ccatctgctg gctcagtcag catagagttt gcccagaatc cttgagtcat atgaggaagg 
                 2280 
               
               
                 ccaacagaaa caaagaattt ctctgcctct tgaaatatcc tttctgcatc ccagccctga 
                 2340 
               
               
                 ttcatcattg catcagtaac atctatgttt ggtttctgtg caaagggaac agtcaaaggg 
                 2400 
               
               
                 tacagatttg tccaaaatct accccacata tcaccaagca aatgggcagg gaggcatcca 
                 2460 
               
               
                 gtggggctga tgtaggaagg gtaggtatcc atcaacttcc tcctcacata ggcatgaaga 
                 2520 
               
               
                 tgctcataca atggcttgat ctctgcgaag gtacgttcta catcttcaat caactggtta 
                 2580 
               
               
                 cggttatagt tgtagccatc tgctccctct gcttcatagt cccctctcca ataatcccca 
                 2640 
               
               
                 tagtcgttat aattgtttgc tcttgccatc tcgtttttca ggaccacata ctcttcatac 
                 2700 
               
               
                 aacggcctca gctgcttgcc aacctcagcc ctccagccct cccatgccca gagcctagag 
                 2760 
               
               
                 ttgtagtctg tgcttgtcgc cattatttca tccaatcctg gctcaagtaa taagcattct 
                 2820 
               
               
                 tgtgggttct ttgggttgca aacttttcca gtactgtaaa tggtgctcat ggtgttcaga 
                 2880 
               
               
                 attgtgttca actgtttgtt cttgtctgct gagagtgctg aagacccact ttgctgaagg 
                 2940 
               
               
                 gcctgtagtt gacgcttgat gatcggagtc tggatttctt gtagtgagaa actttgggca 
                 3000 
               
               
                 gtcttagact gttcttcata aaaggcagac catttggctg cagcctcact catcttttgg 
                 3060 
               
               
                 gcattttctt cagtaatgtt agtattataa ttccaagaag caagtgaact ttgataagac 
                 3120 
               
               
                 aggtcttcag cttcctgatt aaagttgttt aaaaatgtct tggcattttc ctcggtgagg 
                 3180 
               
               
                 gactgagcag tagtaacagc aacaaggctg agaaggagcc aggaggagct ggacatcttt 
                 3240 
               
               
                 ccccgtgcgc caagatccca tccactgagc aactttccct agagtaaaat ctcttcatga 
                 3300 
               
               
                 gtttttctct ctcatcagcc tttgaacttg ggttgggcac tgcttacaac tcagctgaga 
                 3360 
               
               
                 actcaaggaa gtaataatga ctctgcatta tgaaatccaa agcgcatcca tgacaagtcc 
                 3420 
               
               
                 ttggcatcca taggcatggg atcgtggaaa tatgaactcc aaagcaattt taatggttat 
                 3480 
               
               
                 cctggtgaag ttcagcagct ggctccgtgt cctcattccc agagtgtaag agaggagcca 
                 3540 
               
               
                 ctttaaatgg cagggctcat gggcct 
                 3566 
               
               
                   
               
            
           
           
               
               
            
               
                 &lt;210&gt; 
                 10 
               
               
                 &lt;211&gt; 
                 3418 
               
               
                 &lt;212&gt; 
                 DNA 
               
               
                 &lt;213&gt; 
                 
                   Mus musculus 
                 
               
               
                 &lt;400&gt; 
                 10 
               
            
           
           
               
               
            
               
                 agaggagtgt ctaaatctaa taagtattta ttgaggtcgg ctttgtgttc accattgctc 
                   60 
               
               
                 agaccctgtg agaaaataca aacaatttca gagactgggc acccatctca cagatcgagc 
                  120 
               
               
                 attcatctgg aattttccct tactccacaa tgggagaata atggaagtag tttctgacag 
                  180 
               
               
                 gcatgctctg ggttcgtgga gtttcctgtg ctagctgtgt cttgttagag agatgttgat 
                  240 
               
               
                 caagtttgga agagaaaagg tcatggtctc tacctgatgt ctggcttctt tctccgttga 
                  300 
               
               
                 actgcacctc caaagaaggg acagtccatt tcaagtatta ctgtgaacaa attcctacag 
                  360 
               
               
                 tacttgaagt gagaagtgtg aaggcagaag ccacagagca tgacgtcaca cttggattca 
                  420 
               
               
                 agtgaccagc gagcaccatt ctcaacctca acagtcccac cagttcacag cccagatgcc 
                  480 
               
               
                 acaccgaagc caacacttgc ccccaagaga catcatctca tcttctaact ccagttaccc 
                  540 
               
               
                 aaccatatca ccctggatag actgtgaggc caaaggcaca tttccctctt ttatgctaag 
                  600 
               
               
                 ataaacatct ctattaataa acagaagaga tgtcaaatcc ttaggcagaa ataaatactg 
                  660 
               
               
                 aaagcaatgt cagtgaagat gtccactcaa gtctgatgct ctttggacaa cgtttactga 
                  720 
               
               
                 acagatcata gttttgatag ctaatgacat gtgtatactc tcatattttg tgtactatga 
                  780 
               
               
                 agttagcatt tacatacagg aagatgacaa gtgctttgct aaaaggaagt ctgagcatca 
                  840 
               
               
                 tcactgtttt ggaatcctgc attgctttct ccttttccaa tgtccatcga gtcataaggg 
                  900 
               
               
                 ttctcttctc tttttgtttc atttttcttc tttcgacctt tgatcccagt gacaatcagg 
                  960 
               
               
                 atgatgatgc caaccactac cagtgccatc acaacaccaa aaataatcag ccatatggtg 
                 1020 
               
               
                 acaggaggct ggtaaggtgg ctcaagtgtt gggtgaatcc ccagaaactc caggctgtta 
                 1080 
               
               
                 tcattcaggc caaagacatc attgatgcgg ccccgagaca tcctgatggc atcttcaact 
                 1140 
               
               
                 tcacttctag gaatgacatc agacacattt tggggtgagg tgacaaagaa gtagaaggag 
                 1200 
               
               
                 actcttggtt tcaaatcact cactcgtaca tcttcctcta gaaaaggaac tgtctggttt 
                 1260 
               
               
                 ttgattattg aaaaatactt tctcatggca tatgcaacag atgatcggaa caggaacatt 
                 1320 
               
               
                 tcgttgttgg tccattcata tgcattagct ccaagagctg attttaggct tatcctcact 
                 1380 
               
               
                 ttaatgcttt ggtcggcata tgggctccat tcagtgttcc accccacaaa agaatttctg 
                 1440 
               
               
                 ttctgctctt tcagccagtc aaacaacggt tggaagtaat tgagcagtgg ttttacatcc 
                 1500 
               
               
                 atattccttg ctcctaccac attttccaag gctttggtcc agggctctga atttccaaga 
                 1560 
               
               
                 ctcagcatct tgagcaactt ctgcccagct tcagtggaat ttgagatgtc acatttgtgc 
                 1620 
               
               
                 agagaaccat tatacttagc tgcttgacaa agagcttctt gaaactggaa ttggtaaatg 
                 1680 
               
               
                 gtccttgtgt aatatcgaat gaatgagtaa tcattagaaa catggaacag agatgcaggg 
                 1740 
               
               
                 tcacagtatg tttcatcatg aggcagaggc tccaccacac caacgatctc ccgcttcatc 
                 1800 
               
               
                 tcccaccact ttttcatcca ctgctctttg ggaatttcac cccgaaagac catccacctc 
                 1860 
               
               
                 cacttctcta acatgtaagt aaacggtagt gttccaacaa ttgtcaatgc ctgtttcagt 
                 1920 
               
               
                 aggaagttta tctctgtttc gctatcttct tgaaaatcgg atggcagaag accaatggat 
                 1980 
               
               
                 ttcagatgct tgggggtagc tgcagaaagt gacatgattt ctccaacagc ttcatggaac 
                 2040 
               
               
                 ccttcattgg ctccgtttct tagcaggaaa ggttgcctgg catatgccat gtcatattgg 
                 2100 
               
               
                 atgtgtccca tctcgtgatg ggctgtcaag aagttgtcca ttgtgacctt tgtacacatc 
                 2160 
               
               
                 ttgattctga agtctccgtg tcccagatcc caagctgtgg ggtggcagac aactttccgg 
                 2220 
               
               
                 ccatctgctg gctcagtcag catagagttt gcccagaatc cttgagtcat atgaggaagg 
                 2280 
               
               
                 ccaacagaaa caaagaattt ctctgcctct tgaaatatcc tttctgcatc ccagccctga 
                 2340 
               
               
                 ttcatcattg catcagtaac atctatgttt ggtttctgtg caaagggaac agtcaaaggg 
                 2400 
               
               
                 tacagatttg tccaaaatct accccacata tcaccaagca aatgggcagg gaggcatcca 
                 2460 
               
               
                 gtggggctga tgtaggaagg gtaggtatcc atcaacttcc tcctcacata ggcatgaaga 
                 2520 
               
               
                 tgctcataca atggcttgat ctctgcgaag gtacgttcta catcttcaat caactggtta 
                 2580 
               
               
                 cggttatagt tgtagccatc tgctccctct gcttcatagt cccctctcca ataatcccca 
                 2640 
               
               
                 tagtcgttat aattgtttgc tcttgccatc tcgtttttca ggaccacata ctcttcatac 
                 2700 
               
               
                 aacggcctca gctgcttgcc aacctcagcc ctccagccct cccatgccca gagcctagag 
                 2760 
               
               
                 ttgtagtctg tgcttgtcgc cattatttca tccaatcctg gctcaagtaa taagcattct 
                 2820 
               
               
                 tgtgggttct ttgggttgca aacttttcca gtactgtaaa tggtgctcat ggtgttcaga 
                 2880 
               
               
                 attgtgttca actgtttgtt cttgtctgct gagagtgctg aagacccact ttgctgaagg  
                 2940 
               
               
                 gcctgtagtt gacgcttgat gatcggagtc tggatttctt gtagtgagaa actttgggca 
                 3000 
               
               
                 gtcttagact gttcttcata aaaggcagac catttggctg cagcctcact catcttttgg 
                 3060 
               
               
                 gcattttctt cagtaatgtt agtattataa ttccaagaag caagtgaact ttgataagac 
                 3120 
               
               
                 aggtcttcag cttcctgatt aaagttgttt aaaaatgtct tggcattttc ctcggtgagg 
                 3180 
               
               
                 gactgagcag tagtaacagc aacaaggctg agaaggagcc aggaggagct ggacatcttt 
                 3240 
               
               
                 ccccgtgcgc caagatccca tccactgagc aactttccct agagtaaaat ctcttcatga 
                 3300 
               
               
                 gtttttctct ctcatcagcc tttgaacttg ggttgggcac tgagaagaga agaaaaaaaa 
                 3360 
               
               
                 agcagaagaa gaaacagaag agaacagtaa acaagctgct ggaccaatat gaagttaa 
                 3418 
               
               
                   
               
            
           
           
               
               
            
               
                 &lt;210&gt; 
                 11 
               
               
                 &lt;211&gt; 
                 2418 
               
               
                 &lt;212&gt; 
                 DNA 
               
               
                 &lt;213&gt; 
                 
                   Rattus norvegicus 
                 
               
               
                 &lt;400&gt; 
                 11 
               
            
           
           
               
               
            
               
                 ttagaatgaa gtttgagcat catcactgtt ttggaatcct gcgttacttt ctcctttgcc 
                   60 
               
               
                 aatgtccatg gagtcataag gattctcttc tctttttgtt tcatttttct tctttcgacc 
                  120 
               
               
                 tttgatccca gtgacgatca ggataacaat gccaaccact accgttccca tcacgacacc 
                  180 
               
               
                 aaaaataatc agccatatgg tgacaggagg ctcgtaaggt ggcttaagtg ttgggtagat 
                  240 
               
               
                 ccccagaaac tccaggctgt tatcattcag accaaaaata tcattgatac ggccccgaga 
                  300 
               
               
                 catcctgatg gcctcttcaa cttcacttct gggaatgatg tcagacacat ttttgggtga 
                  360 
               
               
                 agtgacaaag aagttgaagg agactcttgg tttcaaatca ctcacccata cgtctgcctc 
                  420 
               
               
                 cccaaaagga actgtctggt tcttttccct tgaaaaatac tctctcatgg cataggcaac 
                  480 
               
               
                 agatgatcgg aataggtaca tttcgttgtc ggtccattca tacgcatttt tcccaagagc 
                  540 
               
               
                 tgattttagg cttatcctca ctttaatgct ttggtcggca tatgggctcc agtcagtgct 
                  600 
               
               
                 ccaccccaca gtcgaattcc tgttctgctc tttcagccag acaaacaatg gttggaagta 
                  660 
               
               
                 attgagcagt ggttttacat ccatattcct tgatcctacc acattttcca aggctagggt 
                  720 
               
               
                 ccagggccct gagtttccaa gactcagcat attgagcaac ttctgcccag cttcagtgga 
                  780 
               
               
                 atttgagatg tcacatttgt gtagtgggcc atcatgttta gctgcttgac aaagagcttc 
                  840 
               
               
                 ttgaaactgg aattgataaa tggtccttgt gtaatatcga atgaatgagt aatcattaga 
                  900 
               
               
                 gacatggaac agagatgcag ggtcacagta tgtttcatca tgaggcagag gctccaccac 
                  960 
               
               
                 accaacgatc tcccgcttca tctcccacca ctttttggtc cactgttctc tgggaatttt 
                 1020 
               
               
                 atcctgaaag accatccacc tccacttctc taacatgtaa gtaaatggca gcgttccaac 
                 1080 
               
               
                 aattgtcaat gcctgtttga gtaggaagtt tatttctgtt tcattgtctt cttgaaaatt 
                 1140 
               
               
                 ggatggcaga agaccaatag atttcaaatg tttgggggta gctgcagaaa gtgacatgat 
                 1200 
               
               
                 ttctccaacg gcttcatgga acccttcatt ggctccgttt cttagcagga aaggttgctt 
                 1260 
               
               
                 ggcatatgcc atgtcatatt ggatgtgtcc catctcatga tgggctgtca agaagttgtc 
                 1320 
               
               
                 cattgtgacc tttgtgcaca tcttgattct gaagtctcca tgtcccagat cccaagctgt 
                 1380 
               
               
                 ggggtggcag acaactttcc ggtcatctcc tggctcagtc agcatggagt ttgtccagaa 
                 1440 
               
               
                 tcccggagtc atttgaggaa ggccaacaga aacgaagaac ttctctgcct ctttaaatat 
                 1500 
               
               
                 tctttctgca tcccagctct gattcaccat tgcatcagta acatctatgt ttggtttctg 
                 1560 
               
               
                 aagaaaggga gtagtcaaag ggtacagatt tgtccaaaac ctaccccaca tatcaccaag 
                 1620 
               
               
                 caaatgagca gggaggcatc cagtagggct gatgtaagaa gggtacactt ccatcaactt 
                 1680 
               
               
                 cgttctcaca taggcatgaa gttgctcata caacggtttg atctctttga aggtattttc 
                 1740 
               
               
                 tacgtcttcg atcaactggt ttcggttgta gttgtaacct tctactccct ctgcttcata 
                 1800 
               
               
                 atcccctcgc caataatccc catagtcttc ataattgttt gctcttgcca tctcattttt 
                 1860 
               
               
                 caggaccaca tactcttcat ataacggcct cagctgcttg ccgacctcag ccctccagcc 
                 1920 
               
               
                 ctcccaagcc cagagcctac gattgtagtc tgtgcttgtt gccattattt cgtccaatcc 
                 1980 
               
               
                 tggttcaagt aaaaaacatt cttgtggatt cattgagttg caaacttttc cagtactgta 
                 2040 
               
               
                 aatggtgctc atggtgttta gaattgtgtt caactgtttg ttcttgtctg gtgacagcgc 
                 2100 
               
               
                 tgaagacccg ctctgctgaa gggccttcag ttgacgcttg atggtcgcat tctgaatttc 
                 2160 
               
               
                 ttgtagtgag aaattttggg cgatcttgga ctgttcttca taaaaggcag accatttggc 
                 2220 
               
               
                 cgcagcctcg ttcatctttt gggcattctc ctccgtaatg ttggtgttgt aattccaaga 
                 2280 
               
               
                 agcaagtgaa ctttgataag acaggtcttc agcttcctgg ttaaacttgt ttaaaaagct 
                 2340 
               
               
                 ctcggccttt tcctcgatga gggactgagc agtagcaaca gcaacaaggc tgagaaggag 
                 2400 
               
               
                 ccagcaggag cttgacat 
                 2418 
               
               
                   
               
            
           
           
               
               
            
               
                 &lt;210&gt; 
                 12 
               
               
                 &lt;211&gt; 
                 3589 
               
               
                 &lt;212&gt; 
                 DNA 
               
               
                 &lt;213&gt; 
                 
                   Macaca fascicularis 
                 
               
               
                 &lt;400&gt; 
                 12 
               
            
           
           
               
               
            
               
                 gagtgtgcaa atctagcatt tattgagtgt ctgctttgtg ctcagcacta ctcaaacact 
                   60 
               
               
                 gtgggcaaat acaaaagatt taagagactg gatacccact tcagagggtg aacatacagt 
                  120 
               
               
                 tggaatttcc attcactcca cagttagaga acagtggaaa tgagtttcta tcaggcatgc 
                  180 
               
               
                 tctgggagtt cacggaggcc cctggcgtta gttttgtgtt atcagggaga tgttgatcaa 
                  240 
               
               
                 gcagcttgga agtgaaaaag caatgtcctc tacctgttcc ctggattctt ctctctttag 
                  300 
               
               
                 acctgtcacc ttgaagaaag cgtgatctat tccaaacatc actgtaggca aatcaccata 
                  360 
               
               
                 gtacttggat taagttaatg tgaagaaagt atgtcatcct taacaatgga ttaaatgaaa 
                  420 
               
               
                 gtgaactagt gcatgccatt ctcaatcctt gcagctgtac cagttcccag gcactgtcct 
                  480 
               
               
                 tacaagtgat ccatccatat tccatacaag gtccaatact tgctcccatg atatatcctt 
                  540 
               
               
                 gttttcaact tcagaaattc aacagacatt tacatttaga ttatccctga acagcccgtg 
                  600 
               
               
                 agaccaaata cacactttcc ctaatacact ctaatacaaa atccttatta ggaaacagaa 
                  660 
               
               
                 cagaagtcaa atccagagac agaaataaat actgaaagca atgtcaatga agatgctctc 
                  720 
               
               
                 tccttgacca tgttgtcttt ggacaatttt tttctgaaca gattcatagt tttgacattt 
                  780 
               
               
                 aatgatatat ttatcatctc atatgttcga taccatgaaa ttaacattta cctacaacaa 
                  840 
               
               
                 aatcacctca agagaaaaaa catagatttt tctaaaagga ggtctgaaca tcatcagtgt 
                  900 
               
               
                 tttggaatcc tggattattt tctcctttgt taatatcgat ggaggcataa ggattttctt 
                  960 
               
               
                 cacttcttgc ttgatttttc ttctttcgat ctctgatccc agtgaagatc aggacgacaa 
                 1020 
               
               
                 tgccagccac tatcactccc atcacaactc caaaaacaat tagccatgtg gtaacggggg 
                 1080 
               
               
                 actggtaagg aggtgcaagt gttgtctgta tccccagaaa ctccaggctg ttgtcattca 
                 1140 
               
               
                 gacggaaagc atcattgata cggctcctgg agatcctgat ggcctcttca acttcagttc 
                 1200 
               
               
                 taggaatgat gtcagacaca tttttaggtg cagtgacata gaaattaaag gagattcttg 
                 1260 
               
               
                 gtttcaaatc agcaactcgc acatcctcct ccccaaaaag aatcgtctga tgtttgattt 
                 1320 
               
               
                 ctaaaaagta cgtcctcatg gcatatgcaa cagatgatcg gaacaggtac atttcattgt 
                 1380 
               
               
                 cgttccattc atatgcttta tctccaagag ctgattttaa gcttatcctc actttgatgc 
                 1440 
               
               
                 tttggtcagc atacggactc cagtcggtac tccatcccac aaaagaattc ttgttctggt 
                 1500 
               
               
                 ctttcagcca ggtaaacaag ggctcaaagt agttgagcag tggtcttaca ttcatgttct 
                 1560 
               
               
                 ttgctcctac aacattttcc aatgctaggg tccagggttc tgattttcca agcttcagca 
                 1620 
               
               
                 tattgagcaa tttctgtcca gcttctgtag agtttgagat gtcacatttg tgcagagggc 
                 1680 
               
               
                 cttcgtgttt agctgcttga caaagtgctt cttgaaactg gaattggtaa agggtccttg 
                 1740 
               
               
                 tgtaatatcg aatgaatgag taatcattag aaacatggaa cagagatgcg gggtcacagt 
                 1800 
               
               
                 atgtttcatc atggggcaca ggttccacca ccccaactat ctctcgcttc atctcccacc 
                 1860 
               
               
                 actttttcat ccactggtct ttgggaattt cacctttaaa gaccatccac ctccacttct 
                 1920 
               
               
                 ctaacatgta agtaaatggc agagtcccaa caatcgtgag tgcttgtttg agcaggaagt 
                 1980 
               
               
                 ttatttctgt ttcattgtct tcttgaaaat caggtgacag aagaccaatg gattttaaat 
                 2040 
               
               
                 gcttaggtgt ggctgcagaa agtgacatga tttccccaac agcttcatgg aatccttcat 
                 2100 
               
               
                 tagctccatt tcttagcaga aaaggttgtg cagcatatgc catatcatat tggatatgcc 
                 2160 
               
               
                 ccatctcatg atgagctgtc aggaagtcgt ccattgtcac ctttgtgcac ataatgatcc 
                 2220 
               
               
                 tgaagtcacc cttccccagg tcccaagctg tggggtggca gactactttc tgaacatttc 
                 2280 
               
               
                 ctggatcagt tagcatggaa ttttcccaga atccttgagt catattagga agaccaacag 
                 2340 
               
               
                 atacaaagaa cttctcggcc tccttgaata ttctctgtgc attccaggcc tggttcacca 
                 2400 
               
               
                 ttgcatcagt aacatctatg tttggtttct gtccaaaggg aactgtcaaa gagtacagat 
                 2460 
               
               
                 ttgtccaaaa tctaccccac atatcaccaa gcaaatgagc aggaaggcat ccagttggac 
                 2520 
               
               
                 taatatagga agggtaggca ttcatcaact ttgccctcac ataggcatga agatgttcat 
                 2580 
               
               
                 ataatggttt aatctcttca aaggtacgtt ccacatcttc aatcaactgg tcgcggttgt 
                 2640 
               
               
                 agtcatagcc atctaccccg tttacttcat agtttcctct ccaataatcc ccatagtcct 
                 2700 
               
               
                 tataatgatt tgctcttgcc atctcatttt tcaagaccac atactcttca tataatggcc 
                 2760 
               
               
                 tcagctgctt gccgacctca gatctccagc cttcccaagc ccagagcctc tcattgtagt 
                 2820 
               
               
                 ctaaactctt ttccattatt tcattcaaac ctggatcaag taataagcat tcctggggat 
                 2880 
               
               
                 tatttgggtt acaaactttt ccagtactgt agatggtgct cattgtattt agaattgtgt 
                 2940 
               
               
                 tcaaccgttt gctcttgtct tctgagagca ctgaagaccc attttgctga agagcctgca 
                 3000 
               
               
                 actgaagctt gactgtgaga ttctgaattt cttgcagtgg atacatttgg gcaagtgtgg 
                 3060 
               
               
                 actgttcttt taaaaaggca gaccattttt ccccagcatt attcatgttt tggacattct 
                 3120 
               
               
                 cttcagtaat attggtgtta taattccaag aagcaagtga actttgatag aacaggtctt 
                 3180 
               
               
                 cggcttcgtg gttaaacttg tccaaaaatg tcttggcctg ttcctcaatg gtggactgag 
                 3240 
               
               
                 cagcagttac agcaacaagg ctgagaagga gccaggaaga gcctgacatc gtcccctgtg 
                 3300 
               
               
                 agccaagatc acatccactg aatgactttc cctagactaa aacctcctca tgagattttc 
                 3360 
               
               
                 tctcttatca gcctttgaac ttgggttggg cactattcaa aactcagtca aggtcacgtg 
                 3420 
               
               
                 gaagtaagaa agcctccata cagtatgaaa tccaaagagt gtctacgtca actcctgatt 
                 3480 
               
               
                 ctctgtagcc acaggatcac aacaatatag aattcaaaga gattttagtg gttatcttgt 
                 3540 
               
               
                 taaatttcag acgcgagctc agtgtcctca ttactagagt gtatgtatg 
                 3589