Patent Publication Number: US-2022228144-A1

Title: SERPIN FAMILY F MEMBER 2 (SERPINF2) 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/US2020/044400, filed on Jul. 31, 2020, which claims the benefit of priority to U.S. Provisional Patent Application No. 62/881,556, filed on Aug. 1, 2019. The entire contents of each of the foregoing applications are incorporated herein by reference. 
    
    
     SEQUENCE LISTING 
     The instant application contains a Sequence Listing which has been submitted electronically in ASCII format and is hereby incorporated by reference in its entirety. Said ASCII copy, created on Jan. 14, 2022, is named 121301_09702_SL.txt and is 157,441 bytes in size. 
     BACKGROUND OF THE INVENTION 
     SERPINF2, also referred to as A2AP, is a member of the serine proteinase inhibitor (serpin) superfamily. SERPINF2 is a plasma protease inhibitor that inhibits proteins such as plasmin, trypsin, matriptase-3/TMPRSS7, and chymotrypsin. Since its original description, α2-antiplasmin (A2AP) or α2-plasmin inhibitor (SERPINF2) has been known as the primary inhibitor of plasmin. A2AP rapidly inactivates plasmin, forming covalent plasmin-A2AP (protease-serpin) complexes, whose presence in the blood is a marker of plasmin generation in clinical states associated with plasminogen activation. Activated factor XIII crosslinks A2AP to fibrin during fibrin formation. Crosslinking of A2AP to the fibrin significantly enhances the resistance of fibrin to degradation by plasmin. 
     Increased levels of A2AP are associated with increased risk of developing disorders associated with thrombosis, e.g., venous thrombosis, venous thromboembolism, deep vein thrombosis, pulmonary embolism, prosthetic valve thrombosis, post-thrombotic syndrome, atherothrombosis, heart attack, stroke, fibrinolytic disorders, hypofibrinolysis, disfibrinolysis, ischemic stroke, atrial fibrillation, myocardial infarction, peripheral arterial disease, dynamic thrombosis, coronary artery disease, microvascular thrombosis, ischemic brain injury, brain swelling, brain hemorrhage and death after thromboembolic stroke. 
     Thrombosis is the formation of a blood clot within a blood vessel. It prevents blood from flowing normally through the circulatory system. When a blood clot forms in the veins, it is known as venous thromboembolism. This can cause deep vein thrombosis and pulmonary embolisms. When a clot forms in the arteries, it is called atherothrombosis, which can lead to heart attack and stroke. 
     The common treatment for thrombosis and is typically non-selective anti-coagulant therapy. Unfortunately, however, the lack of specificity of such therapies can lead to excessive bleeding. Accordingly, there is a need in the art for more effective treatments for disorders associated with thrombosis, e.g., plasminogen deficiency, venous thrombosis, venous thromboembolism, deep vein thrombosis, pulmonary embolism, prosthetic valve thrombosis, post-thrombotic syndrome, atherothrombosis, heart attack, stroke, fibrinolytic disorders, hypofibrinolysis, disfibrinolysis, ischemic stroke, atrial fibrillation, myocardial infarction, peripheral arterial disease, dynamic thrombosis, coronary artery disease, microvascular thrombosis, ischemic brain injury, brain swelling, brain hemorrhage and death after thromboembolic stroke. 
     SUMMARY OF THE INVENTION 
     The present invention provides iRNA compositions which affect the RNA-induced silencing complex (RISC)-mediated cleavage of RNA transcripts of a gene encoding serpin family F member 2 (SERPINF2). The SERPINF2 may be within a cell, e.g., a cell within a subject, such as a human subject. 
     In an aspect, the invention provides a double stranded ribonucleic acid (dsRNA) agent for inhibiting expression of serpin family F member 2 (SERPINF2) in a cell, wherein the dsRNA agent comprises a sense strand and an antisense strand, wherein the sense strand comprises at least 15 contiguous nucleotides differing by no more than 3 nucleotides from the nucleotide sequence of SEQ ID NO:1 and the antisense strand comprises at least 15 contiguous nucleotides differing by no more than 3 nucleotides from the nucleotide sequence of SEQ ID NO:2. 
     In an aspect, the invention provides a double stranded ribonucleic acid (dsRNA) agent for inhibiting expression of serpin family F member 2 (SERPINF2), wherein the dsRNA agent comprises a sense strand and an antisense strand forming a double stranded region, wherein the sense strand comprises at least 15 contiguous nucleotides from any one of the nucleotide sequence of nucleotides 299-321, 580-602, 674-696, 754-776, 969-991, 2123-2145, 2242-2264, or 2244-2266 of SEQ ID NO:1, and the antisense strand comprises at least 15 contiguous nucleotides from the corresponding nucleotide sequence of SEQ ID NO:2. 
     In certain embodiments, the sense strand comprises at least 21 contiguous nucleotides of any one of the nucleotide sequence of nucleotides 299-321, 580-602, 674-696, 754-776, 969-991, 2123-2145, 2242-2264, or 2244-2266 of SEQ ID NO:1. 
     In certain embodiments, the antisense strand comprises at least 19 contiguous nucleotides from any one of the antisense strand nucleotide sequences of a duplex selected from the group consisting of AD-202054.2, AD-202129.2, AD-202191.2, AD-203169.2, AD-203171.2, AD-204882.2, AD-205458.2, AD-206288.2, and AD-329749. 
     In certain embodiments, the sense strand comprises at least 19 contiguous nucleotides from any one of the sense strand nucleotide sequences of a duplex selected from the group consisting of AD-202054.2, AD-202129.2, AD-202191.2, AD-203169.2, AD-203171.2, AD-204882.2, AD-205458.2, AD-206288.2, and AD-329749. 
     In certain embodiments, the sense and antisense strands comprise nucleotide sequences of any one of the duplexes selected from the group consisting of AD-202054.2, AD-202129.2, AD-202191.2, AD-203169.2, AD-203171.2, AD-204882.2, AD-205458.2, AD-206288.2, and AD-329749. 
     In certain embodiments, the dsRNA agent comprises at least one modified nucleotide. In certain embodiments, substantially all of the nucleotides of the sense strand and substantially all of the nucleotides of the antisense strand comprise a modification. In certain embodiments, all of the nucleotides of the sense strand and all of the nucleotides of the antisense strand comprise a modification. In certain embodiments, at least one of the modified nucleotides is selected from the group of 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, 2′-hydroxly-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 phosphorothioate group, a nucleotide comprising a methylphosphonate group, a nucleotide comprising a 5′-phosphate, a nucleotide comprising a 5′-phosphate mimic, a thermally destabilizing nucleotide, a glycol modified nucleotide (GNA), and a 2-O—(N-methylacetamide) modified nucleotide; and combinations thereof. In certain embodiments, the modifications on the nucleotides are selected from the group consisting of LNA, HNA, CeNA, 2′-methoxyethyl, 2′-O-alkyl, 2′-O-allyl, 2′-C-allyl, 2′-fluoro, 2′-deoxy, 2′-hydroxyl, GNA, and combinations thereof. In certain embodiments, the modifications on the nucleotides are 2′-O-methyl or 2′-fluoro modifications. In certain embodiments, at least one of the modified nucleotides is selected from the group consisting of a deoxy-nucleotide, a 2′-O-methyl modified nucleotide, a 2′-fluoro modified nucleotide, a 2′-deoxy-modified nucleotide, a glycol modified nucleotide (GNA), vinyl-phosphonate (VP), and a 2-O—(N-methylacetamide) modified nucleotide; and combinations thereof. In certain embodiments, at least one of the nucleotide modification is a thermally destabilizing nucleotide modification. In certain embodiments, the thermally destabilizing nucleotide modification is selected from the group consisting of an abasic modification; a mismatch with the opposing nucleotide in the duplex; and destabilizing sugar modification, a 2′-deoxy modification, an acyclic nucleotide, an unlocked nucleic acids (UNA), and a glycerol nucleic acid (GNA) 
     In certain embodiments, the double stranded region is 19-30 nucleotides in length. In certain embodiments, the double stranded region is 19-25 nucleotide pairs in length. In certain embodiments, the double stranded region is 19-23 nucleotides in length. In certain embodiments, the double stranded region is 23-27 nucleotides in length. In certain embodiments, the double stranded region is 21-23 nucleotides in length. In certain embodiments, each strand of the dsRNA agent is independently no more than 30 nucleotides in length. In certain embodiments, at least one strand of the dsRNA agent comprises a 3′ overhang of at least 1 nucleotide. In certain embodiments, at least one strand of the dsRNA agent comprises a 3′ overhang of at least 2 nucleotides. 
     In certain embodiments, dsRNA agent further comprises a ligand. In certain embodiments, the ligand is conjugated to the 3′ end of the sense strand of the dsRNA agent. In certain embodiments, the ligand is one or more GalNAc derivatives attached through a monovalent, bivalent, or trivalent branched linker. In certain embodiments, the ligand is an N-acetylgalactosamine (GalNAc) derivative, e.g., wherein the ligand is 
     
       
         
         
             
             
         
       
     
     In certain embodiments, the dsRNA agent is conjugated to the ligand as shown in the following schematic 
     
       
         
         
             
             
         
       
     
     and, wherein X is O or S, e.g., wherein the X is O. 
     In certain embodiments, the dsRNA agent further comprises at least one phosphorothioate or methylphosphonate internucleotide linkage. In certain embodiments, the phosphorothioate or methylphosphonate internucleotide linkage is at the 3′-terminus of one strand. In certain embodiments, the strand is the antisense strand. In certain embodiments, the strand is the sense strand. In certain embodiments, the phosphorothioate or methylphosphonate internucleotide linkage is at the 5′-terminus of one strand. In certain embodiments, the strand is the antisense strand. In certain embodiments, the strand is the sense strand. In certain embodiments, the phosphorothioate or methylphosphonate internucleotide linkage is at the both the 5′- and 3′-terminus of one strand. In certain embodiments, the strand is the antisense strand. 
     In certain embodiments, the dsRNA agent at the 1 position of the 5′-end of the antisense strand of the duplex comprises a base pair that is an AU base pair. 
     In certain embodiments, the sense strand comprises 5′-UGUAGUGAUUUUUAUGCCACA-3′ (SEQ ID NO: 9) and the antisense strand comprises 5′-UGUGGCAUAAAAAUCACUACAAA-′3 (SEQ ID NO: 10). In certain embodiments, the sense strand comprises 5′-usgsuaguGfaUfUfUfuuaugccaca-3′ (SEQ ID NO: 11) and the antisense strand comprises 5′-VPusGfsuggCfaUfAfaaaaUfcAfcuacasasa-′3 (SEQ ID NO: 12), wherein a, g, c, and u are 2′-O-methyl (2′-OMe) A, G, C, and U; Af, Gf, Cf, and Uf are 2′-fluoro A, G, C, U; VP is vinyl-phosphonate; and s is a phosphorothioate linkage. In certain embodiments, the sense strand consists of 5′-usgsuaguGfaUfUfUfuuaugccacaL96-3′ (SEQ ID NO: 13) and the antisense strand consists of 5′-VPusGfsuggCfaUfAfaaaaUfcAfcuacasasa-′3 (SEQ ID NO: 14), wherein a, g, c, and u are 2′-O-methyl (2′-OMe) A, G, C, and U; Af, Gf, Cf, and Uf are 2′-fluoro A, G, C, U; VP is vinyl-phosphonate; L96 is N-[tris(GalNAc-alkyl)-amidodecanoyl)]-4-hydroxyprolinol; and s is a phosphorothioate linkage. 
     In certain embodiments, the sense strand comprises 5′-UGUAGUGAUUUUUAUGCCACA-3′ (SEQ ID NO: 15) and the antisense strand comprises 5′-UGUGGCAUAAAAAUCACUACAAA-‘3 (SEQ ID NO: 16). In certain embodiments, the sense strand comprises 5’-usgsuaguGfaUfUfUfuuaugccaca-3′ (SEQ ID NO: 17) and the antisense strand comprises 5′-usGfsuggCfaUfAfaaaaUfcAfcuacasasa-′3 (SEQ ID NO: 18), wherein a, g, c, and u are 2′-O-methyl (2′-OMe) A, G, C, and U; Af, Gf, Cf, and Uf are 2′-fluoro A, G, C, U; and s is a phosphorothioate linkage. In certain embodiments, the sense strand consists of 5′-usgsuaguGfaUfUfUfuuaugccacaL96-3′ (SEQ ID NO: 19) and the antisense strand consists of 5′-usGfsuggCfaUfAfaaaaUfcAfcuacasasa-′3 (SEQ ID NO: 20), wherein a, g, c, and u are 2′-O-methyl (2′-OMe) A, G, C, and U; Af, Gf, Cf, and Uf are 2′-fluoro A, G, C, U; L96 is N-[tris(GalNAc-alkyl)-amidodecanoyl)]-4-hydroxyprolinol; and s is a phosphorothioate linkage. 
     In an aspect, the invention provides a double-stranded ribonucleic acid (dsRNA) agent for inhibiting expression of SERPINF2, wherein said dsRNA agent comprises a sense strand and an antisense strand, wherein the sense strand consists of the nucleotide sequence of 5′-usgsuaguGfaUfUfUfuuaugccaca-3′ (SEQ ID NO: 21) and the antisense strand consists of the nucleotide sequence of 5′-VPusGfsuggCfaUfAfaaaaUfcAfcuacasasa-3′ (SEQ ID NO: 22), wherein a, g, c, and u are 2′-O-methyl (2′-OMe) A, G, C, and U; Af, Gf, Cf, and Uf are 2′-fluoro A, G, C, U; VP is vinyl-phosphonate; and s is a phosphorothioate linkage, and wherein a ligand is conjugated to the 3′ end of the sense strand, and wherein the ligand has the following structure: 
     
       
         
         
             
             
         
       
     
     In an aspect, the invention provides a double-stranded ribonucleic acid (dsRNA) agent for inhibiting expression of SERPINF2, wherein said dsRNA agent comprises a sense strand and an antisense strand, wherein the sense strand consists of 5′-usgsuaguGfaUfUfUfuuaugccacaL96-3′ (SEQ ID NO: 23) and the antisense strand consists of 5′-usGfsuggCfaUfAfaaaaUfcAfcuacasasa-′3 (SEQ ID NO: 24), wherein a, g, c, and u are 2′-O-methyl (2′-OMe) A, G, C, and U; Af, Gf, Cf, and Uf are 2′-fluoro A, G, C, U; VP is vinyl-phosphonate; and s is a phosphorothioate linkage, and wherein a ligand is conjugated to the 3′ end of the sense strand, and wherein the ligand has the following structure: 
     
       
         
         
             
             
         
       
     
     In an aspect, the invention provides a cell containing the dsRNA agent of the invention. 
     In an aspect, the invention provides a pharmaceutical composition for inhibiting expression of a gene encoding SERPINF2 comprising the dsRNA agent of the invention. In certain embodiments, the pharmaceutical composition comprises the dsRNA agent and a lipid formulation. 
     In an aspect, the invention provides a method of inhibiting expression of an SERPINF2 gene in a cell, the method comprising contacting the cell with the dsRNA agent of the invention or the pharmaceutical composition of the invention, thereby inhibiting expression of the SERPINF2 gene in the cell. 
     In certain embodiments, the cell is within a subject. In certain embodiments, the subject is a human. In certain embodiments, the subject has been diagnosed with an SERPINF2-associated disorder. 
     In certain embodiments, the SERPINF2-associated disorder is a disorder associated with thrombosis, e.g., plasminogen deficiency, venous thrombosis, venous thromboembolism, deep vein thrombosis, pulmonary embolism, prosthetic valve thrombosis, post-thrombotic syndrome, atherothrombosis, heart attack, stroke, fibrinolytic disorders, hypofibrinolysis, disfibrinolysis, ischemic stroke, atrial fibrillation, myocardial infarction, peripheral arterial disease, dynamic thrombosis, coronary artery disease, microvascular thrombosis, ischemic brain injury, brain swelling, brain hemorrhage and death after thromboembolic stroke. 
     In certain embodiments, contacting the cell with the dsRNA agent inhibits the expression of SERPINF2 by at least 50%, 60%, 70%, 80%, 90%, 95% (e.g., as compared to the level of expression of SERPINF2 prior to first contacting the cell with the dsRNA agent; e.g., prior to administration of a first dose of the dsRNA agent to the subject). In certain embodiments, inhibiting expression of SERPINF2 decreases an SERPINF2 protein level in a subject serum sample(s) by at least 50%, 60%, 70%, 80%, 90%, or 95%, e.g., as compared to the level of expression of SERPINF2 prior to first contacting the cell with the dsRNA agent. 
     In an aspect, the invention provides a method of treating a an SERPINF2-associated disorder in a subject, comprising administering to the subject the dsRNA agent or the pharmaceutical composition of the invention, thereby treating the SERPINF2-associated disorder in the subject. In certain embodiments, the subject is human. 
     In certain embodiments of the invention, the dsRNA agent is administered at a dose of about 0.01 mg/kg to about 50 mg/kg. In certain embodiments, the dsRNA agent is administered to the subject subcutaneously. In certain embodiments, the level of SERPINF2 is measured in the subject. In certain embodiments, the level of SERPINF2 in the subject is an SERPINF2 protein level in a subject blood or serum sample. 
     In certain embodiments, an additional therapeutic agent for treatment of a SERPINF2-associated disorder is administered to the subject. In certain embodiments, the additional therapeutic agent is an anticoagulant. In some embodiments, the anticoagulant includes heparin, enoxaparin (Lovenox), dalteparin (Fragmin), fondaparinux (Arixtra), warfarin (Coumadin, Jantoven), dabigatran (Pradaxa), rivaroxaban (Xarelto), apixaban (Eliquis), edoxaban (Savaysa), argatroban or any combination thereof. In some embodiments, the additional therapeutic agent includes a thrombolytic. In certain embodiments, the thrombolytic includes antistreplase (Eminase), tissue plasminogen activator (tPA), urokinase-type plasminogen activator (uPA), or any combination thereof. In some embodiments, the additional therapeutic agent is an immunosuppressant. In certain embodiments, the immunosuppresant includes corticosteroid, azathioprine, cyclosporine A, or any combination thereof. In some embodiments, the additional therapeutic agent is hormone replacement therapy. In certain embodiments, the hormone replacement therapy includes estrogen, gestagen, androgen or any combination thereof. In some embodiments, the additional therapeutic agent is an antibiotic. In some embodiments, the additional therapeutic agent is an antihistamine agent. In some embodiments, the additional therapeutic agent is an mast cell stablizer. In certain embodiments, the mast cell stabilizer includes cromoglicic acid (Cromolyn), lodoxamide (Alomide), or any combination thereof. In some embodiments, the additional therapeutic agent is an anti-proliferative agent. In some embodiments, the additional therapeutic agent is an oral contraceptive. In some embodiments, the additional therapeutic agent is a fresh frozen plasma or a plasminogen concentrate. In some embodiments, the additional therapeutic agent is hyaluronidase. In some embodiments, the additional therapeutic agent is alpha chymotrypsin. In certain embodiment, the additional therapeutic agent is a filter inserted into a large vein that prevents clots that break loose from lodging in the patient&#39;s lungs. In certain embodiments, the additional therapeutic agent is selected from the group consisting of an anticoagulant, a SERPINF2 inhibitor, a thrombin inhibitor. 
     The invention also provides uses of the dsRNA agents and the pharmaceutical compositions provided herein for treatment of an SERPINF2-associated disorder. In certain embodiments, the uses include any of the methods provided by the invention. 
     The invention provides kits or pharmaceutical compositions comprising a dsRNA agent of the invention. In certain embodiments, the invention provides kits for practicing a method of the invention. 
    
    
     
       BRIEF DESCRIPTION OF THE DRAWINGS 
         FIG. 1  schematically depicts the nucleotide sequences of dsRNA duplexes targeting SERPINF2 that were subcutaneously administered to mice (n=3 per group) as a single 2 mg/kg dose (SEQ ID NOs 507-522, respectively, in order of appearance). 
         FIG. 2  is a graph showing SERPINF2 mRNA levels in mice (n=3 per group) treated with a single 2 mg/kg dose of the indicated dsRNA duplexes, on day 0. SERPINF2 levels are shown relative to control levels detected with PBS treatment. 
         FIG. 3  is a graph showing SERPINF2 mRNA levels in mice (n=3 per group) treated with a single 0.3 mg/kg, 3 mg/kg, or 10 mg/kg dose of AD-329749 dsRNA on day 0. SERPINF2 levels are shown relative to control levels detected with PBS treatment. 
         FIG. 4A  is a graph showing reduced fibrin deposition levels in electrolytic injury model mice (n=8 per group) treated with a 10 mg/kg dose of AD-203169 (A2AP-siRNA). 
         FIG. 4B  is a graph showing significant reduction in thrombus size in electrolytic injury model mice (n=8 per group) treated with a 10 mg/kg dose of AD-203169 (A2AP-siRNA). 
     
    
    
     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 a SERPINF2 gene. The 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 (SERPINF2 gene) in mammals. 
     The iRNAs of the invention have been designed to target the huma SERPINF2 gene, including portions of the gene that are conserved in the SERPINF2 orthologs of other mammalian species. Without intending to be limited by theory, it is believed that a combination or sub-combination of the foregoing properties and the specific target sites or the specific modifications in these iRNAs confer to the iRNAs of the invention improved efficacy, stability, potency, durability, and safety. 
     Accordingly, the present invention also provides methods for treating and preventing a SERPINF2-associated disorder, e.g., a disorder associated with thrombosis, e.g., plasminogen deficiency, venous thrombosis, venous thromboembolism, deep vein thrombosis, pulmonary embolism, prosthetic valve thrombosis, post-thrombotic syndrome, atherothrombosis, heart attack, stroke, fibrinolytic disorders, hypofibrinolysis, disfibrinolysis, ischemic stroke, atrial fibrillation, myocardial infarction, peripheral arterial disease, dynamic thrombosis, coronary artery disease, microvascular thrombosis, ischemic brain injury, brain swelling, brain hemorrhage and death after thromboembolic stroke, using iRNA compositions which effect the RNA-induced silencing complex (RISC)-mediated cleavage of RNA transcripts of a SERPINF2 gene. 
     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 a SERPINF2 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 transcript of a SERPINF2 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 mRNAs of the corresponding gene (SERPINF2 gene) in mammals. Using in vitro and in vivo assays, the present inventors have demonstrated that iRNAs targeting a SERPINF2 gene can mediate RNAi, resulting in significant inhibition of expression of SERPINF2. Inhibition of expression of SERPINF2 in such a subject will prevent or treat development of a SERPINF2-associated disorder, e.g., a disorder associated with thrombosis, e.g., plasminogen deficiency, venous thrombosis, venous thromboembolism, deep vein thrombosis, pulmonary embolism, prosthetic valve thrombosis, post-thrombotic syndrome, atherothrombosis, heart attack, stroke, fibrinolytic disorders, hypofibrinolysis, disfibrinolysis, ischemic stroke, atrial fibrillation, myocardial infarction, peripheral arterial disease, dynamic thrombosis, coronary artery disease, microvascular thrombosis, ischemic brain injury, brain swelling, brain hemorrhage and death after thromboembolic stroke. Thus, methods and compositions including these iRNAs are useful for preventing and treating a subject susceptible to or diagnosed with a SERPINF2-associated disorder, e.g., a disorder associated with thrombosis, such as plasminogen deficiency. The methods and compositions herein are useful for reducing the level of SERPINF2 in a subject. 
     The following detailed description discloses how to make and use compositions containing iRNAs to inhibit the expression of a SERPINF2 gene as well as compositions, uses, and methods for treating subjects that would benefit from reduction of the expression of a SERPINF2 gene, e.g., subjects susceptible to or diagnosed with a SERPINF2-associated disorder, e.g., a disorder associated with thrombosis, such as plasminogen deficiency. 
     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. For example, “sense strand or antisense strand” is understood as “sense strand or antisense strand or sense strand and antisense strand.” 
     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. 
     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, “serpin family F member 2,” used interchangeably with the term “SERPINF2” refers to the well-known gene and polypeptide, also known in the art as Serpin Family F Member 2; Alpha-2-Plasmin Inhibitor; Alpha-2-Antiplasmin; Serine (Or Cysteine) Proteinase Inhibitor, Clade F (Alpha-2 Antiplasmin, Pigment Epithelium Derived Factor), Member 2; Serpin Peptidase Inhibitor, Clade F (Alpha-2 Antiplasmin, Pigment Epithelium Derived Factor), Member 2; Alpha-2-AP; Serpin F2; PLI; AAP; Plasmin Inhibitor; ALPHA-2-PI; Alpha-2-PI; A2AP; and API. 
     The term “SERPINF2” includes huma SERPINF2, the amino acid and complete coding sequence of which may be found in for example, GenBank Accession No. GI:124053441 (NM_000934.3; SEQ ID NO:1);  Macaca fascicularis  SERPINF2, the amino acid and complete coding sequence of which may be found in for example, GenBank Accession No. GI: 982302431 (XM_005582451.2; SEQ ID NO:3); mouse ( Mus musculus ) SERPINF2, the amino acid and complete coding sequence of which may be found in for example, GenBank Accession No. GI: 292781734 (NM_008878.2; SEQ ID NO:5); and rat ( Rattus norvegicus ) SERPINF2 the amino acid and complete coding sequence of which may be found in for example, for example GenBank Accession No. GI:58865361 (NM_001011892.1; SEQ ID NO:7). 
     Additional examples of SERPINF2 mRNA sequences are readily available using publicly available databases, e.g., GenBank, UniProt, OMIM, and the  Macaca  genome project web site. 
     The term“SERPINF2,” as used herein, also refers to naturally occurring DNA sequence variations of the SERPINF2 gene, such as a single nucleotide polymorphism (SNP) in the SERPINF2 gene. Exemplary SNPs may be found in the dbSNP database available at https://www.ncbi.nlm nih.gov/snp/?term=serpinf2. 
     As used herein, “target sequence” refers to a contiguous portion of the nucleotide sequence of an mRNA molecule formed during the transcription of a SERPINF2 gene, including mRNA that is a product of RNA processing of a primary transcription product. The target portion of the sequence will be at least long enough to serve as a substrate for iRNA-directed cleavage at or near that portion of the nucleotide sequence of an mRNA molecule formed during the transcription of a SERPINF2 gene. In one embodiment, the target sequence is within the protein coding region of SERPINF2. 
     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. 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. iRNA directs the sequence-specific degradation of mRNA through a process known as RNA interference (RNAi). The iRNA modulates, e.g., inhibits, the expression of a SERPINF2 gene in a cell, e.g., a cell within a subject, such as a mammalian subject. 
     In one embodiment, an RNAi agent of the invention includes a single stranded RNA that interacts with a target RNA sequence, e.g., a SERPINF2 target 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 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). Thus, in one aspect the invention relates to a single stranded RNA (siRNA) generated within a cell and which promotes the formation of a RISC complex to effect silencing of the target gene, i.e., a SERPINF2 gene. Accordingly, the term “siRNA” is also used herein to refer to an iRNA as described above. 
     In certain embodiments, the RNAi agent may be a single-stranded siRNA (ssRNAi) 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 siRNAs 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 certain embodiments, an “iRNA” for use in the compositions, uses, and methods of the invention is a double stranded RNA and is referred to herein as a “double stranded RNA 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., a SERPINF2 gene. In some embodiments of the invention, 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, the majority of nucleotides of each strand of a dsRNA molecule are ribonucleotides, but as described in detail herein, each or both strands can also include one or more non-ribonucleotides, e.g., a deoxyribonucleotide or a modified nucleotide. In addition, as used in this specification, an “iRNA” may include ribonucleotides with chemical modifications; an iRNA 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 modified nucleobase, or any combination thereof. 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 invention 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 “iRNA” or “RNAi agent” for the purposes of this specification and claims. 
     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 19 to 36 base pairs in length, e.g., about 19-30 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 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 least 2, 3, 4, 5, 6, 7, 8, 9, 10, 20, 23 or more unpaired nucleotides. 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. 
     Where the two substantially complementary strands of a dsRNA are comprised by separate RNA molecules, those molecules need not be, 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 certain embodiments, an iRNA agent of the invention is a dsRNA, each strand of which comprises 19-23 nucleotides, that interacts with a target RNA sequence, e.g., a SERPINF2 gene, to direct cleavage of the target RNA. 
     In some embodiments, an iRNA of the invention is a dsRNA of 24-30 nucleotides that interacts with a target RNA sequence, e.g., a SERPINF2 target 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 double stranded iRNA. 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 certain embodiments, the antisense 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 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, 10-25 nucleotides, 10-20 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 extended 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. 
     “Blunt” or “blunt end” means that there are no unpaired nucleotides at that end of the double stranded RNA agent, i.e., no nucleotide overhang. A “blunt ended” double stranded RNA agent is double stranded over its entire length, i.e., no nucleotide overhang at either end of the molecule. The RNAi agents of the invention include RNAi agents with no nucleotide overhang at one end (i.e., agents with one overhang and one blunt end) or with no nucleotide overhangs at either end. 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., a SERPINF2 mRNA. 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., a SERPINF2 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, or 3 nucleotides of the 5′- or 3′-end of the iRNA. In some embodiments, a double stranded RNA agent of the invention includes a nucleotide mismatch in the antisense strand. In some embodiments, a double stranded RNA agent of the invention includes a nucleotide mismatch in the sense 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. 
     The term “sense strand” or “passenger strand” as used herein, refers to the strand of an iRNA 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. Such conditions can, for example, be stringent conditions, where stringent conditions can include: 400 mM NaCl, 40 mM PIPES pH 6.4, 1 mM EDTA, 50° C. or 70° C. for 12-16 hours followed by washing (see, e.g., “ Molecular Cloning: A Laboratory Manual , Sambrook, et al. (1989) Cold Spring Harbor Laboratory Press). Other 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 an iRNA, 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 double stranded RNA 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) refers to a polynucleotide that is substantially complementary to a contiguous portion of the mRNA of interest (e.g., an mRNA encoding a SERPINF2 gene). For example, a polynucleotide is complementary to at least a part of a SERPINF2 mRNA if the sequence is substantially complementary to a non-interrupted portion of an mRNA encoding a SERPINF2 gene. 
     Accordingly, in some embodiments, the sense strand polynucleotides and the antisense polynucleotides disclosed herein are fully complementary to the target SERPINF2 sequence. In other embodiments, the sense strand polynucleotides or the antisense polynucleotides disclosed herein are substantially complementary to the target SERPINF2 sequence and comprise a contiguous nucleotide sequence which is at least 80% complementary over its entire length to the equivalent region of the nucleotide sequence of any one of SEQ ID NOs:1 and 2, or a fragment of any one of SEQ ID NOs:1 and 2, such as at least 90%, or 95% complementary; or 100% complementary. 
     Accordingly, in some embodiments, the antisense strand polynucleotides disclosed herein are fully complementary to the target SERPINF2 sequence. In other embodiments, the antisense strand polynucleotides disclosed herein are substantially complementary to the target SERPINF2 sequence and comprise a contiguous nucleotide sequence which is at least about 90% complementary over its entire length to the equivalent region of the nucleotide sequence of SEQ ID NO:1, or a fragment of SEQ ID NO:1, such as about 90%, or about 95%, complementary. In certain embodiments, the fragment of SEQ ID NO: 1 is selected from the group of nucleotides 2231-2253, 2214-2236, 674-696, 2244-2266, 802-824, 682-704, 759-781, 2230-2252, 675-697, 2242-2264, 578-600, 2257-2279, 1084-1106, 1083-1105, 576-598, 760-782, 755-777, 2215-2237, 1091-1113, 580-602, 963-985, 1601-1623, 2123-2145, 754-776, 2250-2272, 1081-1103, 978-1000, 990-1012, 977-999, 579-601, 2239-2261, 1663-1685, 576-598, 1082-1104, 577-599, 1813-1835, 1086-1108, 1824-1846, 1823-1845, 2218-2240, 997-1019, 1832-1854, 840-862, 678-700, 1814-1836, 989-1011, 681-703, 676-698, 984-1006, 357-379, 1592-1614, 896-918, 761-783, 964-986, 761-783, 1011-1033, 762-784, 302-324, 766-788, 969-991, 895-917, 1802-1824, 893-915, 998-1020, 970-992, 966-988, 746-768, 301-323, 305-327, 752-774, 311-333, 755-777, 1964-1986, 677-699, 980-1002, 381-403, 974-996, 1801-1823, 2116-2138, 1803-1825, 1965-1987, 704-726, 380-402, 299-321, 691-713, 1800-1822, 967-989, 975-997, 883-905, or 479-501 of SEQ ID NO: 1. 
     In some embodiments, an iRNA of the invention includes an antisense strand that is substantially complementary to the target SERPINF2 sequence and comprises a contiguous nucleotide sequence which is at least about 90% complementary over its entire length to the equivalent region of the nucleotide sequence of any one of the sense strands in Table 2, Table 3, Table 6 or Table 8 or a fragment of any one of the sense strands in Table 2, Table 3, Table 6 or Table 8, such as about 90%, 95%, or 100% 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 SERPINF2 sequence, and wherein the sense strand polynucleotide comprises a contiguous nucleotide sequence which is at least about 90% complementary over its entire length to the equivalent region of the nucleotide sequence of any one of the antisense strands in Table 2, Table 3, Table 6 or Table 8, or a fragment of any one of the antisense strands in Table 2 or Table 3, such as about 90%, 95%, or 100%. 
     In certain embodiments, the sense and antisense strands in Table 2 or Table 3 are selected from duplexes AD-203163.1, AD-203147.1, AD-202129.1, AD-203171.1, AD-202239.1, AD-202137.1, AD-202196.1, AD-203162.1, AD-202130.1, AD-203169.1, AD-202052.1, AD-203184.1, AD-202492.1, AD-202491.1, AD-202050.1, AD-202197.1, AD-202192.1, AD-203148.1, AD-202499.1, AD-202054.1, AD-202372.1, AD-202796.1, AD-206288.1, AD-202191.1, AD-203177.1, AD-202489.1, AD-202387.1, AD-202399.1, AD-202386.1, AD-202053.1, AD-203166.1, AD-202827.1, AD-205123.1, AD-202490.1, AD-202051.1, AD-202894.1, AD-202494.1, AD-202905.1, AD-202904.1, AD-203151.1, AD-202406.1, AD-202913.1, AD-202277.1, AD-202133.1, AD-202895.1, AD-202398.1, AD-202136.1, AD-202131.1, AD-202393.1, AD-201919.1, AD-202787.1, AD-205414.1, AD-205279.1, AD-202373.1, AD-202198.1, AD-202420.1, AD-202199.1, AD-204885.1, AD-205284.1, AD-205458.1, AD-205413.1, AD-206058.1, AD-205411.1, AD-202407.1, AD-205459.1, AD-205455.1, AD-205264.1, AD-204884.1, AD-204888.1, AD-205270.1, AD-204894.1, AD-205273.1, AD-206218.1, AD-202132.1, AD-202389.1, AD-204944.1, AD-205463.1, AD-206057.1, AD-206281.1, AD-206059.1, AD-206219.1, AD-205227.1, AD-204943.1, AD-204882.1, AD-205214.1, AD-206056.1, AD-205456.1, AD-205464.1, AD-205401.1, AD-205034.1, and AD-329749. 
     In general, an “iRNA” includes ribonucleotides with chemical modifications. Such modifications may include all types of modifications disclosed herein or known in the art. Any such modifications, as used in a dsRNA molecule, are encompassed by “iRNA” for the purposes of this specification and claims. 
     In an aspect of the invention, an agent for use in the methods and compositions of the invention is a single-stranded antisense oligonucleotide molecule that inhibits a target mRNA via an antisense inhibition mechanism. The single-stranded antisense oligonucleotide 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 oligonucleotide molecule may be about 14 to about 30 nucleotides in length and have a sequence that is complementary to a target sequence. For example, the single-stranded antisense oligonucleotide molecule may comprise a sequence that is at least about 14, 15, 16, 17, 18, 19, 20, or more contiguous nucleotides from any one of the antisense sequences described herein. 
     The phrase “contacting a cell with an iRNA,” such as a dsRNA, as used herein, includes contacting a cell by any possible means. Contacting a cell with an iRNA includes contacting a cell in vitro with the iRNA or contacting a cell in vivo with the iRNA. The contacting may be done directly or indirectly. Thus, for example, the iRNA may be put into physical contact with the cell by the individual performing the method, or alternatively, the iRNA 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 iRNA. Contacting a cell in vivo may be done, for example, by injecting the iRNA into or near the tissue where the cell is located, or by injecting the iRNA into another area, e.g., 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 iRNA may contain or be coupled to a ligand, e.g., GalNAc, that directs the iRNA to a site of interest, e.g., the liver. 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 iRNA and subsequently transplanted into a subject. 
     In certain embodiments, contacting a cell with an iRNA includes “introducing” or “delivering the iRNA into the cell” by facilitating or effecting uptake or absorption into the cell. Absorption or uptake of an iRNA can occur through unaided diffusion or active cellular processes, or by auxiliary agents or devices. Introducing an iRNA into a cell may be in vitro or in vivo. For example, for in vivo introduction, iRNA 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 “lipid nanoparticle” or “LNP” is a vesicle comprising a lipid layer encapsulating a pharmaceutically active molecule, such as a nucleic acid molecule, e.g., an iRNA or a plasmid from which an iRNA 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), a non-primate (such as 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 an embodiment, the subject is a human, such as a human being treated or assessed for a disease or disorder that would benefit from reduction in SERPINF2 expression; a human at risk for a disease or disorder that would benefit from reduction in SERPINF2 expression; a human having a disease or disorder that would benefit from reduction in SERPINF2 expression; or human being treated for a disease or disorder that would benefit from reduction in SERPINF2 expression as described herein. The diagnostic criteria for a SERPINF2-associated disorder, e.g., a disorder associated with thrombosis, e.g., plasminogen deficiency, venous thrombosis, venous thromboembolism, deep vein thrombosis, pulmonary embolism, prosthetic valve thrombosis, post-thrombotic syndrome, atherothrombosis, heart attack, stroke, fibrinolytic disorders, hypofibrinolysis, disfibrinolysis, ischemic stroke, atrial fibrillation, myocardial infarction, peripheral arterial disease, dynamic thrombosis, coronary artery disease, microvascular thrombosis, ischemic brain injury, brain swelling, brain hemorrhage and death after thromboembolic stroke, are provided below. 
     In some embodiments, the subject is a female human. In other embodiments, the subject is a male human. In certain embodiments, the subject is overweight or obese, e.g., a subject that suffers from central obesity. In certain embodiments, the subject is sedentary. In certain embodiments, the subject is pregnant. 
     As used herein, the terms “treating” or “treatment” refer to a beneficial or desired result, such as reducing at least one sign or symptom of a SERPINF2-associated disorder, e.g., thrombosis, or deposits of fibrin-rich “pseudomembranes” (woody or ligneous membranes) that impair normal tissue and organ function, e.g., ligneous conjunctivitis, i.e., peudomembranes or ligneous lesions that affect the eyes; or ligneous lesions that affect the gingiva, oropharynx, middle ear, renal collecting system, respiratory tract, central nervous system, skin, and female reproductive tract in the subject. Treatment also includes a reduction of one or more sign or symptoms associated with unwanted SERPINF2 expression, e.g., stabilization of active SERPINF2; diminishing the extent of unwanted SERPINF2 activation or stabilization; amelioration or palliation of unwanted SERPINF2 activation or stabilization. “Treatment” can also mean prolonging survival as compared to expected survival in the absence of treatment. Non-limiting examples of SERPINF2-associated diseases or disorders include plasminogen deficiency, venous thrombosis, venous thromboembolism, deep vein thrombosis, pulmonary embolism, prosthetic valve thrombosis, post-thrombotic syndrome, atherothrombosis, heart attack, stroke, fibrinolytic disorders, hypofibrinolysis, disfibrinolysis, ischemic stroke, atrial fibrillation, myocardial infarction, peripheral arterial disease, dynamic thrombosis, coronary artery disease, microvascular thrombosis, ischemic brain injury, brain swelling, brain hemorrhage and death after thromboembolic stroke. 
     The term “lower” in the context of the level of SERPINF2 gene expression or SERPINF2 protein production 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 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, or 95%, or below the level of detection for the detection method in a relevant cell or tissue, e.g., a liver cell or tissue, an eye cell or tissue, or other subject sample, e.g., blood or serum derived therefrom, or urine. 
     As used herein, “prevention” or “preventing,” when used in reference to a disease or disorder, that would benefit from a reduction in expression of a SERPINF2 gene or production of SERPINF2 protein, e.g., in a subject susceptible to a SERPINF2-associated disorder due to, e.g., aging, genetic factors, such as plasminogen deficiency, hormone changes, diet, and a sedentary lifestyle. In certain embodiments, the disease or disorder is a disorder associated with thrombosis, e.g., plasminogen deficiency, venous thrombosis, venous thromboembolism, deep vein thrombosis, pulmonary embolism, prosthetic valve thrombosis, post-thrombotic syndrome, atherothrombosis, heart attack, stroke, fibrinolytic disorders, hypofibrinolysis, disfibrinolysis, ischemic stroke, atrial fibrillation, myocardial infarction, peripheral arterial disease, dynamic thrombosis, coronary artery disease, microvascular thrombosis, ischemic brain injury, brain swelling, brain hemorrhage and death after thromboembolic stroke, in a subject. The failure to develop a SERPINF2-associated disorder, e.g., a disorder associated with thrombosis, e.g., plasminogen deficiency, venous thrombosis, venous thromboembolism, deep vein thrombosis, pulmonary embolism, prosthetic valve thrombosis, post-thrombotic syndrome, atherothrombosis, heart attack, stroke, fibrinolytic disorders, hypofibrinolysis, disfibrinolysis, ischemic stroke, atrial fibrillation, myocardial infarction, peripheral arterial disease, dynamic thrombosis, coronary artery disease, microvascular thrombosis, ischemic brain injury, brain swelling, brain hemorrhage and death after thromboembolic stroke, or a delay in the time to develop by months or years is considered effective prevention. Prevention may require administration of more than one dose of the iRNA agent. 
     The term “serpin family F member 2-associated disease” includes a disease, disorder or condition that would benefit from reduction in SERPINF2 expression, e.g., a blood clotting disorder, e.g., by inhibing SERPINF2 expression, SERPINF2 is unable to inactivate plamin and also fails to enhance the resistance of fibrin to degration by plasimin, thus inhibiting a blood clot from forming and growing. In some embodiments, the SERPINF2-associated disease or disorder is a disease or disorder associated with thrombosis. Non-limiting examples of serpin family F member 2-associated diseases include plasminogen deficiency, venous thrombosis, venous thromboembolism, deep vein thrombosis, pulmonary embolism, prosthetic valve thrombosis, post-thrombotic syndrome, atherothrombosis, heart attack, stroke, fibrinolytic disorders, hypofibrinolysis, disfibrinolysis, ischemic stroke, atrial fibrillation, myocardial infarction, peripheral arterial disease, dynamic thrombosis, coronary artery disease, microvascular thrombosis, ischemic brain injury, brain swelling, brain hemorrhage and death after thromboembolic stroke. 
     In one embodiment, a serpin family F member 2-associated disease is plasminogen deficiency. Plasminogen deficiency is a disorder that results in development of fibrin-rich “pseudomembranes” that impair normal tissue and organ function. Plasminogen deficiency is an inherited clotting disorder caused by mutations in the PLG gene, which encodes plasminogen. Normally, plasminogen is converted by plasminogen activator into plasmin, which breaks down fibrin. Fibrin is the main protein involved in blood clots and is important for wound healing, creating the framework for normal tissue to grow back. Normally, excess fibrin is broken down when no longer needed, and the new, more flexible normal tissue takes its place. 
     Mutations in the PLG gene can decrease the amount of plasminogen that is produced, its function, or both. When the mutations affect plasminogen levels as well as the activity of the protein, affected individuals are said to have type I plasminogen deficiency or hypoplasminogenemia. Subjects with mutations that result in normal levels of plasminogen with reduced activity are said to have type II congenital plasminogen deficiency or dysplasminogenemia. 
     A reduction in functional plasminogen results in less plasmin to break down fibrin, leading to a buildup of fibrin. The excess fibrin and the resulting inflammation of the tissue result in the inflamed woody growths on the mucous membranes. The most common symptom of this disorder is ligneous conjunctivitis. This occurs when a buildup of fibrin causes inflammation of the conjunctiva, which are the mucous membranes that protect the white part of the eye (the sclera) and line the eyelids, leading to thick, woody (ligneous), inflamed growths. These growths can lead to tearing of the cornea, scarring, and vision loss. Other physiologic systems are affected including the gingiva, oropharynx, middle ear, renal collecting system, respiratory tract, central nervous system, skin, and female reproductive tract. 
     This excess fibrin and reduced fibrinolysis in subjects having plasminogen deficiency are associated with an increased thrombotic tendency or development of abnormal clots in the blood vessels, e.g., the veins, and include thrombophlebitis (red inflamed veins), pulmonary embolism, and stroke. 
     In one embodiment, an serpin family F member 2-associated disease is post-thrombotic syndrome. “Post-thrombotic syndrome” or “PTS” is a chronic complication of deep venous thrombosis (DVT), which includes symptoms and signs of chronic venous insufficiency that develop following DVT. A combination of reflux due to valvular incompetence and venous hypertension due to thrombotic obstruction is thought to underlie the pathophysiology of PST. Symptoms and signs of post-thrombotic syndrome may include leg pain, leg heaviness, vein dilation, edema, skin pigmentation, and venous ulcers. 
     A “therapeutically-effective amount” or “prophylactically effective amount” also includes an amount of an RNAi agent that produces some desired effect at a reasonable benefit/risk ratio applicable to any treatment. The iRNA employed in the methods of the present invention 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. Such carriers are known in the art. Pharmaceutically acceptable carriers include carriers for administration by injection. 
     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, 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 the liver (e.g., whole liver or certain segments of liver or certain types of cells in the liver, such as, e.g., hepatocytes). In certain embodiments, samples may be derived from the eye (e.g., the conjunctiva, the mucous membrane, the eyelid, the sclera, or certain types of cells in the eye, e.g., a ganglion cell, a bipolar neuron, a rod cell or a cone cell.) In some embodiments, a “sample derived from a subject” refers to urine obtained from the subject. A “sample derived from a subject” can refer to blood or blood derived serum or plasma from the subject. 
     I. iRNAs of the Invention 
     The present invention provides iRNAs which inhibit the expression of a SERPINF2 gene. In preferred embodiments, the iRNA includes double stranded ribonucleic acid (dsRNA) molecules for inhibiting the expression of a SERPINF2 gene in a cell, such as a cell within a subject, e.g., a mammal, such as a human susceptible to developing a SERPINF2-associated disorder, e.g., a disorder associated with thrombosis, e.g., plasminogen deficiency, venous thrombosis, venous thromboembolism, deep vein thrombosis, pulmonary embolism, prosthetic valve thrombosis, post-thrombotic syndrome, atherothrombosis, heart attack, stroke, fibrinolytic disorders, hypofibrinolysis, disfibrinolysis, ischemic stroke, atrial fibrillation, myocardial infarction, peripheral arterial disease, dynamic thrombosis, coronary artery disease, microvascular thrombosis, ischemic brain injury, brain swelling, brain hemorrhage and death after thromboembolic stroke. The dsRNAi agent includes an antisense strand having a region of complementarity which is complementary to at least a part of an mRNA formed in the expression of a SERPINF2 gene. The region of complementarity is about 19-30 nucleotides in length (e.g., about 30, 29, 28, 27, 26, 25, 24, 23, 22, 21, 20, or 19 nucleotides in length). Upon contact with a cell expressing the SERPINF2 gene, the iRNA inhibits the expression of the SERPINF2 gene (e.g., a human, a primate, a non-primate, or a rat SERPINF2 gene) by at least about 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 flow cytometric techniques. In preferred embodiments, inhibition of expression is determined by the qPCR method provided in the examples, especially in Example 2 with the siRNA at a 10 nM concentration in an appropriate organism cell line provided therein. In preferred embodiments, inhibition of expression in vivo is determined by knockdown of the human gene in a rodent expressing the human gene, e.g., a mouse or an AAV-infected mouse expressing the human target gene, e.g., when administered a single dose at 3 mg/kg at the nadir of RNA expression. RNA expression in liver is determined using the PCR methods provided in Example 2. 
     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. The target sequence can be derived from the sequence of an mRNA formed during the expression of a SERPINF2 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 19 to 30 base pairs in length. Similarly, the region of complementarity to the target sequence is 19 to 30 nucleotides in length. 
     In some embodiments, the dsRNA is about 19 to about 23 nucleotides in length, or about 25 to about 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 in length may 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 19 to about 30 base pairs, e.g., about 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. 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, an iRNA agent useful to target SERPINF2 gene 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., 1-4, 2-4, 1-3, 2-3, 1, 2, 3, or 4 nucleotides. dsRNAs having at least one nucleotide overhang can have superior inhibitory properties relative to their blunt-ended counterparts. 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 an antisense or sense strand of a dsRNA. 
     A dsRNA can be synthesized by standard methods known in the art. Double stranded RNAi 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. Similarly, single-stranded oligonucleotides of the invention can be prepared using solution-phase or solid-phase organic synthesis or both. 
     In an aspect, a dsRNA of the invention includes at least two nucleotide sequences, a sense sequence and an anti-sense sequence. The sense strand is selected from the group of sequences provided in Table 2, Table 3, Table 6 and Table 8, and the corresponding antisense strand of the sense strand is selected from the group of sequences of Table 2, Table 3, Table 6 and Table 8. 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 a SERPINF2 gene. As such, in this aspect, a dsRNA will include two oligonucleotides, where one oligonucleotide is described as the sense strand in Table 2, Table 3, Table 6 or Table 8, and the second oligonucleotide is described as the corresponding antisense strand of the sense strand in Table 2, Table 3, Table 6 or Table 8. In certain embodiments, the substantially complementary sequences of the dsRNA are contained on separate oligonucleotides. In other embodiments, the substantially complementary sequences of the dsRNA are contained on a single oligonucleotide. In certain embodiments, the sense or antisense strand from Table 2 or Table 3 is selected from AD-203163.1, AD-203147.1, AD-202129.1, AD-203171.1, AD-202239.1, AD-202137.1, AD-202196.1, AD-203162.1, AD-202130.1, AD-203169.1, AD-202052.1, AD-203184.1, AD-202492.1, AD-202491.1, AD-202050.1, AD-202197.1, AD-202192.1, AD-203148.1, AD-202499.1, AD-202054.1, AD-202372.1, AD-202796.1, AD-206288.1, AD-202191.1, AD-203177.1, AD-202489.1, AD-202387.1, AD-202399.1, AD-202386.1, AD-202053.1, AD-203166.1, AD-202827.1, AD-205123.1, AD-202490.1, AD-202051.1, AD-202894.1, AD-202494.1, AD-202905.1, AD-202904.1, AD-203151.1, AD-202406.1, AD-202913.1, AD-202277.1, AD-202133.1, AD-202895.1, AD-202398.1, AD-202136.1, AD-202131.1, AD-202393.1, AD-201919.1, AD-202787.1, AD-205414.1, AD-205279.1, AD-202373.1, AD-202198.1, AD-202420.1, AD-202199.1, AD-204885.1, AD-205284.1, AD-205458.1, AD-205413.1, AD-206058.1, AD-205411.1, AD-202407.1, AD-205459.1, AD-205455.1, AD-205264.1, AD-204884.1, AD-204888.1, AD-205270.1, AD-204894.1, AD-205273.1, AD-206218.1, AD-202132.1, AD-202389.1, AD-204944.1, AD-205463.1, AD-206057.1, AD-206281.1, AD-206059.1, AD-206219.1, AD-205227.1, AD-204943.1, AD-204882.1, AD-205214.1, AD-206056.1, AD-205456.1, AD-205464.1, AD-205401.1, AD-205034.1, and AD-329749. 
     It will be understood that, although the sequences in Table 2 are not described as modified or conjugated sequences, the RNA of the iRNA of the invention e.g., a dsRNA of the invention, may comprise any one of the sequences set forth in Table 2, that are modified, or the sequences of Table 3 that are conjugated. In other words, the invention encompasses dsRNA of Table 2, Table 3, Table 6 and Table 8 which are un-modified, un-conjugated, modified, or conjugated, as described herein. 
     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.,  EMBO  2001, 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 in any one of Table 2, Table 3, Table 6 and Table 8, dsRNAs described herein can include at least one strand of a length of minimally 21 nucleotides. It can be reasonably expected that shorter duplexes having one of the sequences of Table 2, Table 3, Table 6 and Table 8 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 19, 20, or more contiguous nucleotides derived from one of the sequences of Table 2, Table 3, Table 6 and Table 8, and differing in their ability to inhibit the expression of a SERPINF2 gene by not more than about 5, 10, 15, 20, 25, or 30% inhibition from a dsRNA comprising the full sequence, are contemplated to be within the scope of the present invention. 
     In addition, the RNAs provided in Table 2, Table 3, Table 6 and Table 8 identify a site(s) in a SERPINF2 transcript that is susceptible to RISC-mediated cleavage. As such, the present invention further features iRNAs that target within one of these sites. As used herein, an iRNA is said to target within a particular site of an RNA transcript if the iRNA promotes cleavage of the transcript anywhere within that particular site. Such an iRNA will generally include at least about 19 contiguous nucleotides from one of the sequences provided in Table 2, Table 3, Table 6 and Table 8 coupled to additional nucleotide sequences taken from the region contiguous to the selected sequence in a SERPINF2 gene. 
     II. Modified iRNAs of the Invention 
     In certain embodiments, the RNA of the iRNA of the invention e.g., a dsRNA, is un-modified, and does not comprise, e.g., chemical modifications or conjugations known in the art and described herein. In other embodiments, the RNA of an iRNA of the invention, e.g., a dsRNA, is chemically modified to enhance stability or other beneficial characteristics. In certain embodiments of the invention, substantially all of the nucleotides of an iRNA of the invention are modified. In other embodiments of the invention, all of the nucleotides of an iRNA or substantially all of the nucleotides of an iRNA are modified, i.e., not more than 5, 4, 3, 2, or 1 unmodified nucleotides are present in a strand of the iRNA. 
     The nucleic acids featured in the invention 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 iRNA compounds 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 iRNA 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. 
     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. No. 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. 
     Suitable RNA mimetics are contemplated for use in iRNAs provided herein, 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 in which 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 US 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 iRNAs of the invention are described in, for example, in Nielsen et al.,  Science,  1991, 254, 1497-1500. 
     Some embodiments featured in the invention 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 iRNAs, 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 , NO 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 an iRNA, or a group for improving the pharmacodynamic properties of an iRNA, 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′-O-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 ) and 2′-fluoro (2′-F). Similar modifications can also be made at other positions on the RNA of an iRNA, 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. iRNAs can also have sugar mimetics such as cyclobutyl moieties in place of the pentofuranosyl sugar. Representative US 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 iRNA 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 deoxy-thymine (dT), 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., Angewandte Chemie, International Edition, 1991, 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 invention. These include 5-substituted pyrimidines, 6-azapyrimidines and N-2, N-6 and O-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. 
     The RNA of an iRNA 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). 
     In some embodiments, the RNA of an iRNA can also be modified to include one or more bicyclic sugar moieties. 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 invention 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′-CH 2 —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 invention 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 invention 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′-(CH 2 )—S-2′; 4′-(CH 2 ) 2 —O-2′ (ENA); 4′-CH(CH 3 )—O-2′ (also referred to as “constrained ethyl” or “cEt”) and 4′-CH(CH 2 OCH 3 )—O-2′ (and analogs thereof; see, e.g., U.S. Pat. No. 7,399,845); 4′-C(CH 3 )(CH 3 )—O-2′ (and analogs thereof; see e.g., U.S. Pat. No. 8,278,283); 4′-CH 2 —N(OCH 3 )-2′ (and analogs thereof; see e.g., U.S. Pat. No. 8,278,425); 4′-CH 2 —O—N(CH 3 )-2′ (see, e.g., U.S. Patent Publication No. 2004/0171570); 4′-CH 2 —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′-CH 2 —C(H)(CH 3 )-2′ (see, e.g., Chattopadhyaya et al.,  J. Org. Chem.,  2009, 74, 118-134); and 4′-CH 2 —C(═CH 2 )-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 U.S. patents and U.S. Patent 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). 
     The RNA of an iRNA 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(CH 3 )—O-2′ bridge. In one embodiment, a constrained ethyl nucleotide is in the S conformation referred to herein as “S-cEt.” 
     An iRNA of the invention 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, U.S. Patent Publication No. 2013/0190383; and PCT publication WO 2013/036868, the entire contents of each of which are hereby incorporated herein by reference. 
     In some embodiments, an iRNA of the invention 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 U.S. 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 PCT Publication No. WO 2011/005861. 
     Other modifications of the nucleotides of an iRNA of the invention include a 5′ phosphate or 5′ phosphate mimic, e.g., a 5′-terminal phosphate or phosphate mimic on the antisense strand of an iRNA. Suitable phosphate mimics are disclosed in, for example U.S. Patent Publication No. 2012/0157511, the entire contents of which are incorporated herein by reference. 
     A. Modified iRNAs Comprising Motifs of the Invention 
     In certain aspects of the invention, the double stranded RNA agents of the invention include agents with chemical modifications as disclosed, for example, in WO2013/075035, the entire contents of each of which are incorporated herein by reference. WO2013/075035 provides motifs of three identical modifications on three consecutive nucleotides into a sense strand or antisense strand of a dsRNAi agent, particularly at or near the cleavage site. In some embodiments, the sense strand and antisense strand of the dsRNAi agent may otherwise be completely modified. The introduction of these motifs interrupts the modification pattern, if present, of the sense or antisense strand. The dsRNAi agent may be optionally conjugated with a GalNAc derivative ligand, for instance on the sense strand. 
     More specifically, when the sense strand and antisense strand of the double stranded RNA agent are completely modified to have one or more motifs of three identical modifications on three consecutive nucleotides at or near the cleavage site of at least one strand of a dsRNAi agent, the gene silencing activity of the dsRNAi agent was observed. 
     Accordingly, the invention provides double stranded RNA agents capable of inhibiting the expression of a target gene (i.e., SERPINF2 gene) in vivo. The RNAi agent comprises a sense strand and an antisense strand. Each strand of the RNAi agent may be, for example, 17-30 nucleotides in length, 25-30 nucleotides in length, 27-30 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. 
     The sense strand and antisense strand typically form a duplex double stranded RNA (“dsRNA”), also referred to herein as “dsRNAi agent.” The duplex region of a dsRNAi agent may be, for example, the duplex region can be 27-30 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 19, 20, 21, 22, 23, 24, 25, 26, and 27 nucleotides in length. 
     In certain embodiments, the dsRNAi 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, independently, 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 certain embodiments, the overhang regions can include extended overhang regions as provided above. 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 certain embodiments, the nucleotides in the overhang region of the dsRNAi 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), 2′-O-methoxyethyl-5-methyluridine (Teo), 2′-O-methoxyethyladenosine (Aeo), 2′-O-methoxyethyl-5-methylcytidine (m5Ceo), 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 dsRNAi 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 some embodiments, the overhang is present at the 3′-end of the sense strand, antisense strand, or both strands. In some embodiments, this 3′-overhang is present in the antisense strand. In some embodiments, this 3′-overhang is present in the sense strand. 
     The dsRNAi 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′-end of the sense strand or, alternatively, at the 3′-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 dsRNAi agent 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 certain embodiments, the dsRNAi 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 other embodiments, the dsRNAi 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 other embodiments, the dsRNAi 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 certain embodiments, the dsRNAi 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 certain embodiments, every nucleotide in the sense strand and the antisense strand of the dsRNAi agent, including the nucleotides that are part of the motifs are modified nucleotides. In certain embodiments each residue is independently modified with a 2′-O-methyl or 3′-fluoro, e.g., in an alternating motif. Optionally, the dsRNAi agent further comprises a ligand (preferably GalNAc 3 ). 
     In certain embodiments, the dsRNAi 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′-O-methyl modifications on three consecutive nucleotides at or near the cleavage site. 
     In certain embodiments, the dsRNAi agent comprises sense and antisense strands, wherein the dsRNAi 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 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 dsRNAi 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 dsRNAi agent further comprises a ligand. 
     In certain embodiments, the sense strand of the dsRNAi 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 certain embodiments, the antisense strand of the dsRNAi 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 a dsRNAi agent having a duplex region of 19-23 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; the 10, 11, 12 positions; the 11, 12, 13 positions; the 12, 13, 14 positions; or the 13, 14, 15 positions of the antisense strand, the count starting from the first nucleotide from the 5′-end of the antisense strand, or, the count starting from the first 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 dsRNAi agent from the 5′-end. 
     The sense strand of the dsRNAi 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 some embodiments, the sense strand of the dsRNAi 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 chemistries 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 dsRNAi agent may contain more than one motifs 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 some embodiments, the wing modification on the sense strand or antisense strand of the dsRNAi 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 other embodiments, the wing modification on the sense strand or antisense strand of the dsRNAi 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 dsRNAi 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 dsRNAi 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 some embodiments, every nucleotide in the sense strand and antisense strand of the dsRNAi agent, including the nucleotides that are part of the motifs, 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 0 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 a RNA. For example, a phosphorothioate modification at a non-linking 0 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. For example, 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, CRN, cET, UNA, HNA, CeNA, 2′-methoxyethyl, 2′-O-methyl, 2′-O-allyl, 2′-C-allyl, 2′-deoxy, 2′-hydroxyl, or 2′-fluoro. The strands can contain more than one modification. In one embodiment, each residue of the sense strand and antisense strand is independently modified with 2′-O-methyl or 2′-fluoro. 
     At least two different modifications are typically present on the sense strand and antisense strand. Those two modifications may be the 2′-O-methyl or 2′-fluoro modifications, or others. 
     In certain embodiments, the N a  or N b  comprise modifications of an alternating pattern. The term “alternating motif” 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 dsRNAi agent of the invention 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′ to 3′ of the strand and the alternating motif in the antisense strand may start with “BABABA” from 5′ to 3′ of the strand within the duplex region. As another example, the alternating motif in the sense strand may start with “AABBAABB” from 5′ to 3′ of the strand and the alternating motif in the antisense strand may start with “BBAABBAA” from 5′ to 3′ 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. 
     In some embodiments, the dsRNAi agent comprises the pattern of the alternating motif of 2′-O-methyl modification and 2′-F modification on the sense strand initially has a shift relative to the pattern of the alternating motif of 2′-O-methyl modification and 2′-F modification on the antisense strand initially, i.e., the 2′-O-methyl modified nucleotide on the sense strand base pairs with a 2′-F modified nucleotide on the antisense strand and vice versa. The 1 position of the sense strand may start with the 2′-F modification, and the 1 position of the antisense strand may start with the 2′-O-methyl modification. 
     The introduction of one or more motifs of three identical modifications on three consecutive nucleotides to the sense strand or antisense strand interrupts the initial modification pattern present in the sense strand or antisense strand. This interruption of the modification pattern of the sense or antisense strand by introducing one or more motifs of three identical modifications on three consecutive nucleotides to the sense or antisense strand may enhance the gene silencing activity against the target gene. 
     In some embodiments, when the motif of three identical modifications on three consecutive nucleotides is introduced to any of the strands, the modification of the nucleotide next to the motif is a different modification than the modification of the motif. For example, the portion of the sequence containing the motif is “ . . . N a YYYN b  . . . ,” where “Y” represents the modification of the motif of three identical modifications on three consecutive nucleotide, and “N a ” and “N b ” represent a modification to the nucleotide next to the motif “YYY” that is different than the modification of Y, and where N a  and N b  can be the same or different modifications. Alternatively, N a  or N b  may be present or absent when there is a wing modification present. 
     The iRNA 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, antisense strand, or both strands 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 may contain 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 one embodiment, a double-stranded RNAi agent comprises 6-8 phosphorothioate internucleotide linkages. In some embodiments, the antisense strand comprises two phosphorothioate internucleotide linkages at the 5′-end and two phosphorothioate internucleotide linkages at the 3′-end, and the sense strand comprises at least two phosphorothioate internucleotide linkages at either the 5′-end or the 3′-end. 
     In some embodiments, the dsRNAi agent comprises a phosphorothioate or methylphosphonate internucleotide linkage modification in the overhang region. For example, the overhang region may contain 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 the 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. These terminal three nucleotides may be at the 3′-end of the antisense strand, the 3′-end of the sense strand, the 5′-end of the antisense strand, or the 5′ end of the antisense strand. 
     In some embodiments, the 2-nucleotide overhang is at the 3′-end of the antisense strand, and there are 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. Optionally, the dsRNAi agent may additionally have 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, the dsRNAi agent comprises mismatch(es) with the target, within the duplex, or combinations thereof. The mismatch 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 certain embodiments, the dsRNAi 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 certain embodiments, the nucleotide at the 1 position within the duplex region from the 5′-end in the antisense strand is selected from 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 other embodiments, the nucleotide at the 3′-end of the sense strand is deoxy-thymine (dT) or the nucleotide at the 3′-end of the antisense strand is deoxy-thymine (dT). For example, there is a short sequence of deoxy-thymine nucleotides, for example, two dT nucleotides on the 3′-end of the sense, antisense strand, or both strands. 
     In certain embodiments, the sense strand sequence may be represented by formula (I): 
     
       
         
           
               
               
            
               
                   
                 (I) 
               
               
                   
                 5′ n p -N a -(XXX) i -N b -YYY-N b -(ZZZ) j -N a -n q  3′  
               
            
           
         
       
     
     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 some embodiments, the N a  or N b  comprises modifications of alternating pattern. 
     In some embodiments, the YYY motif occurs at or near the cleavage site of the sense strand. For example, when the dsRNAi 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 first nucleotide, from the 5′-end; or optionally, the count starting at the first 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: 
     
       
         
           
               
               
            
               
                   
                 (Ib) 
               
               
                   
                 5′ n p -N a -YYY-N b -ZZZ-N a -n q  3′; 
               
               
                   
                   
               
               
                   
                 (Ic) 
               
               
                   
                 5′ n p -N a -XXX-N b -YYY-N a -n q  3′; 
               
               
                   
                 or 
               
               
                   
                   
               
               
                   
                 (Id) 
               
               
                   
                 5′ n p -N a -XXX-N b -YYY-N b -ZZZ-N a -n q  3′. 
               
            
           
         
       
     
     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: 
     
       
         
           
               
               
            
               
                   
                 (Ia) 
               
               
                   
                 5′ n p -N a -YYY-N a -n q  3′. 
               
            
           
         
       
     
     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): 
     
       
         
           
               
            
               
                 (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′ 
               
            
           
         
       
     
     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 some embodiments, the N a ′ or N b ′ comprises modifications of alternating pattern. 
     The Y′Y′Y′ motif occurs at or near the cleavage site of the antisense strand. For example, when the dsRNAi agent has a duplex region of 17-23 nucleotides 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 first nucleotide, from the 5′-end; or optionally, the count starting at the first paired nucleotide within the duplex region, from the 5′-end. Preferably, the Y′Y′Y′ motif occurs at positions 11, 12, 13. 
     In certain embodiments, Y′Y′Y′ motif is all 2′-OMe modified nucleotides. 
     In certain embodiments, 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 a ′-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: 
     
       
         
           
               
               
            
               
                   
                 (Ia) 
               
               
                   
                 5′ n p′ -N a′ -Y′Y′Y′-N a′ -n q′  3′. 
               
            
           
         
       
     
     When the antisense strand is represented as formula (IIa), 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, CRN, UNA, cEt, 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 some embodiments, the sense strand of the dsRNAi 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 first nucleotide from the 5′-end, or optionally, the count starting at the first 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 some embodiments the antisense strand may contain Y′Y′Y′ motif occurring at positions 11, 12, 13 of the strand, the count starting from the first nucleotide from the 5′-end, or optionally, the count starting at the first 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 dsRNAi agents for use in the methods of the invention may comprise a sense strand and an antisense strand, each strand having 14 to 30 nucleotides, the iRNA duplex represented by formula (III): 
     
       
         
           
               
            
               
                 (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′ 
               
            
           
         
       
     
     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 an iRNA duplex include the formulas below: 
     
       
         
           
               
            
               
                 (IIIa) 
               
               
                 5′ n p -N a -YYY-N a -n q  3′ 
               
               
                 3′ n p ′-N a ′-Y′Y′Y′-N a ′n q ′ 5′ 
               
               
                   
               
               
                 (IIIb) 
               
               
                 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′ 
               
               
                   
               
               
                 (IIIc) 
               
               
                 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′ 
               
               
                   
               
               
                 (IIId) 
               
               
                 5′ n p -N a -XXX-N b -YYY-N b -ZZZ-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′ 
               
            
           
         
       
     
     When the dsRNAi 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 dsRNAi 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 dsRNAi 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 dsRNAi 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. 
     Each of X, Y, and Z in formulas (III), (IIIa), (IIIb), (IIIc), and (IIId) may be the same or different from each other. 
     When the dsRNAi agent is represented by formula (III), (IIIa), (IIIb), (IIIc), and (IIId), at least one of the Y nucleotides may form a base pair with one of the Y′ nucleotides. Alternatively, at least two of the Y nucleotides form base pairs with the corresponding Y′ nucleotides; or all three of the Y nucleotides all form base pairs with the corresponding Y′ nucleotides. 
     When the dsRNAi agent is represented by formula (IIIb) or (IIId), at least one of the Z nucleotides may form a base pair with one of the Z′ nucleotides. Alternatively, at least two of the Z nucleotides form base pairs with the corresponding Z′ nucleotides; or all three of the Z nucleotides all form base pairs with the corresponding Z′ nucleotides. 
     When the dsRNAi agent is represented as formula (IIIc) or (IIId), at least one of the X nucleotides may form a base pair with one of the X′ nucleotides. Alternatively, at least two of the X nucleotides form base pairs with the corresponding X′ nucleotides; or all three of the X nucleotides all form base pairs with the corresponding X′ nucleotides. 
     In certain embodiments, the modification on the Y nucleotide is different than the modification on the Y′ nucleotide, the modification on the Z nucleotide is different than the modification on the Z′ nucleotide, or the modification on the X nucleotide is different than the modification on the X′ nucleotide. 
     In certain embodiments, when the dsRNAi agent is represented by formula (IIId), the N a  modifications are 2′-O-methyl or 2′-fluoro modifications. In other embodiments, 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 other embodiments, 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 GalNAc derivatives attached through a bivalent or trivalent branched linker (described below). In other embodiments, 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 GalNAc derivatives attached through a bivalent or trivalent branched linker. 
     In some embodiments, when the dsRNAi 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 GalNAc derivatives attached through a bivalent or trivalent branched linker. 
     In some embodiments, the dsRNAi 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 some embodiments, the dsRNAi 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 dsRNAi agents represented by at least one of formulas (III), (IIIa), (IIIb), (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 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. 
     In certain embodiments, an RNAi agent of the invention may contain a low number of nucleotides containing a 2′-fluoro modification, e.g., 10 or fewer nucleotides with 2′-fluoro modification. For example, the RNAi agent may contain 10, 9, 8, 7, 6, 5, 4, 3, 2, 1 or 0 nucleotides with a 2′-fluoro modification. In a specific embodiment, the RNAi agent of the invention contains 10 nucleotides with a 2′-fluoro modification, e.g., 4 nucleotides with a 2′-fluoro modification in the sense strand and 6 nucleotides with a 2′-fluoro modification in the antisense strand. In another specific embodiment, the RNAi agent of the invention contains 6 nucleotides with a 2′-fluoro modification, e.g., 4 nucleotides with a 2′-fluoro modification in the sense strand and 2 nucleotides with a 2′-fluoro modification in the antisense strand. 
     In other embodiments, an RNAi agent of the invention may contain an ultra low number of nucleotides containing a 2′-fluoro modification, e.g., 2 or fewer nucleotides containing a 2′-fluoro modification. For example, the RNAi agent may contain 2, 1 of 0 nucleotides with a 2′-fluoro modification. In a specific embodiment, the RNAi agent may contain 2 nucleotides with a 2′-fluoro modification, e.g., 0 nucleotides with a 2-fluoro modification in the sense strand and 2 nucleotides with a 2′-fluoro modification in the antisense strand. 
     Various publications describe multimeric iRNAs that can be used in the methods of the invention. Such publications include WO2007/091269, U.S. Pat. No. 7,858,769, WO2010/141511, WO2007/117686, WO2009/014887, and WO2011/031520 the entire contents of each of which are hereby incorporated herein by reference. 
     As described in more detail below, the iRNA that contains conjugations of one or more carbohydrate moieties to an iRNA can optimize one or more properties of the iRNA. In many cases, the carbohydrate moiety will be attached to a modified subunit of the iRNA. For example, the ribose sugar of one or more ribonucleotide subunits of a iRNA 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, or 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 iRNA 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 decalin; preferably, the acyclic group is a serinol backbone or diethanolamine backbone. 
     In another embodiment of the invention, an iRNA agent comprises a sense strand and an antisense strand, each strand having 14 to 40 nucleotides. The RNAi agent may be represented by formula (L): 
     
       
         
         
             
             
         
       
     
     In formula (L), B1, B2, B3, B1′, B2′, B3′, and B4′ each are independently a nucleotide containing a modification selected from the group consisting of 2′-O-alkyl, 2′-substituted alkoxy, 2′-substituted alkyl, 2′-halo, ENA, and BNA/LNA. In one embodiment, B1, B2, B3, B1′, B2′, B3′, and B4′ each contain 2′-OMe modifications. In one embodiment, B1, B2, B3, B1′, B2′, B3′, and B4′ each contain 2′-OMe or 2′-F modifications. In one embodiment, at least one of B1, B2, B3, B1′, B2′, B3′, and B4′ contain 2′-O—N-methylacetamido (2′-O-NMA) modification. 
     C1 is a thermally destabilizing nucleotide placed at a site opposite to the seed region of the antisense strand (i.e., at positions 2-8 of the 5′-end of the antisense strand). For example, C1 is at a position of the sense strand that pairs with a nucleotide at positions 2-8 of the 5′-end of the antisense strand. In one example, C1 is at position 15 from the 5′-end of the sense strand. C1 nucleotide bears the thermally destabilizing modification which can include abasic modification; mismatch with the opposing nucleotide in the duplex; and sugar modification such as 2′-deoxy modification or acyclic nucleotide e.g., unlocked nucleic acids (UNA) or glycerol nucleic acid (GNA). In one embodiment, C1 has thermally destabilizing modification selected from the group consisting of: i) mismatch with the opposing nucleotide in the antisense strand; ii) abasic modification selected from the group consisting of: 
     
       
         
         
             
             
         
       
     
     and iii) sugar modification selected from the group consisting of: 
     
       
         
         
             
             
         
       
     
     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. In one embodiment, the thermally destabilizing modification in C1 is a mismatch selected from the group consisting of G:G, G:A, G:U, G:T, A:A, A:C, C:C, C:U, C:T, U:U, T:T, and U:T; and optionally, at least one nucleobase in the mismatch pair is a 2′-deoxy nucleobase. In one example, the thermally destabilizing modification in C1 is GNA or 
     
       
         
         
             
             
         
       
     
     T1, T1′, T2′, and T3′ each independently represent a nucleotide comprising a modification providing the nucleotide a steric bulk that is less or equal to the steric bulk of a 2′-OMe modification. A steric bulk refers to the sum of steric effects of a modification. Methods for determining steric effects of a modification of a nucleotide are known to one skilled in the art. The modification can be at the 2′ position of a ribose sugar of the nucleotide, or a modification to a non-ribose nucleotide, acyclic nucleotide, or the backbone of the nucleotide that is similar or equivalent to the 2′ position of the ribose sugar, and provides the nucleotide a steric bulk that is less than or equal to the steric bulk of a 2′-OMe modification. For example, T1, T1′, T2′, and T3′ are each independently selected from DNA, RNA, LNA, 2′-F, and 2′-F-5′-methyl. In one embodiment, T1 is DNA. In one embodiment, T1′ is DNA, RNA or LNA. In one embodiment, T2′ is DNA or RNA. In one embodiment, T3′ is DNA or RNA.
 
n 1 , n 3 , and q 1  are independently 4 to 15 nucleotides in length.
 
n 5 , q 3 , and q 7  are independently 1-6 nucleotide(s) in length.
 
n 4 , q 2 , and q 6  are independently 1-3 nucleotide(s) in length; alternatively, n 4  is 0.
 
q 5  is independently 0-10 nucleotide(s) in length.
 
n 2  and q 4  are independently 0-3 nucleotide(s) in length.
 
     Alternatively, n 4  is 0-3 nucleotide(s) in length. 
     In one embodiment, n 4  can be 0. In one example, n 4  is 0, and q 2  and q 6  are 1. In another example, n 4  is 0, and q 2  and q 6  are 1, with two phosphorothioate internucleotide linkage modifications within position 1-5 of the sense strand (counting from the 5′-end of the sense strand), 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 of the antisense strand). 
     In one embodiment, n 4 , q 2 , and q 6  are each 1. 
     In one embodiment, n 2 , n 4 , q 2 , q 4 , and q 6  are each 1. 
     In one embodiment, C1 is at position 14-17 of the 5′-end of the sense strand, when the sense strand is 19-22 nucleotides in length, and n 4  is 1. In one embodiment, C1 is at position 15 of the 5′-end of the sense strand 
     In one embodiment, T3′ starts at position 2 from the 5′ end of the antisense strand. In one example, T3′ is at position 2 from the 5′ end of the antisense strand and q 6  is equal to 1. 
     In one embodiment, T1′ starts at position 14 from the 5′ end of the antisense strand. In one example, T1′ is at position 14 from the 5′ end of the antisense strand and q 2  is equal to 1. 
     In an exemplary embodiment, T3′ starts from position 2 from the 5′ end of the antisense strand and T1′ starts from position 14 from the 5′ end of the antisense strand. In one example, T3′ starts from position 2 from the 5′ end of the antisense strand and q 6  is equal to 1 and T1′ starts from position 14 from the 5′ end of the antisense strand and q 2  is equal to 1. 
     In one embodiment, T1′ and T3′ are separated by 11 nucleotides in length (i.e. not counting the T1′ and T3′ nucleotides). 
     In one embodiment, T1′ is at position 14 from the 5′ end of the antisense strand. In one example, T1′ is at position 14 from the 5′ end of the antisense strand and q 2  is equal to 1, and the modification at the 2′ position or positions in a non-ribose, acyclic or backbone that provide less steric bulk than a 2′-OMe ribose. 
     In one embodiment, T3′ is at position 2 from the 5′ end of the antisense strand. In one example, T3′ is at position 2 from the 5′ end of the antisense strand and q 6  is equal to 1, and the modification at the 2′ position or positions in a non-ribose, acyclic or backbone that provide less than or equal to steric bulk than a 2′-OMe ribose. 
     In one embodiment, T1 is at the cleavage site of the sense strand. In one example, T1 is at position 11 from the 5′ end of the sense strand, when the sense strand is 19-22 nucleotides in length, and n 2  is 1. In an exemplary embodiment, T1 is at the cleavage site of the sense strand at position 11 from the 5′ end of the sense strand, when the sense strand is 19-22 nucleotides in length, and n 2  is 1, 
     In one embodiment, T2′ starts at position 6 from the 5′ end of the antisense strand. In one example, T2′ is at positions 6-10 from the 5′ end of the antisense strand, and q 4  is 1. 
     In an exemplary embodiment, T1 is at the cleavage site of the sense strand, for instance, at position 11 from the 5′ end of the sense strand, when the sense strand is 19-22 nucleotides in length, and n 2  is 1; T1′ is at position 14 from the 5′ end of the antisense strand, and q 2  is equal to 1, and the modification to T1′ is at the 2′ position of a ribose sugar or at positions in a non-ribose, acyclic or backbone that provide less steric bulk than a 2′-OMe ribose; T2′ is at positions 6-10 from the 5′ end of the antisense strand, and q 4  is 1; and T3′ is at position 2 from the 5′ end of the antisense strand, and q 6  is equal to 1, and the modification to T3′ is at the 2′ position or at positions in a non-ribose, acyclic or backbone that provide less than or equal to steric bulk than a 2′-OMe ribose. 
     In one embodiment, T2′ starts at position 8 from the 5′ end of the antisense strand. In one example, T2′ starts at position 8 from the 5′ end of the antisense strand, and q 4  is 2. 
     In one embodiment, T2′ starts at position 9 from the 5′ end of the antisense strand. In one example, T2′ is at position 9 from the 5′ end of the antisense strand, and q 4  is 1. 
     In one embodiment, BF is 2′-OMe or 2′-F, q 1  is 9, T1′ is 2′-F, q 2  is 1, B2′ is 2′-OMe or 2′-F, q 3  is 4, T2′ is 2′-F, q 4  is 1, B3′ is 2′-OMe or 2′-F, q 5  is 6, T3′ is 2′-F, q 6  is 1, B4′ is 2′-OMe, and q 7  is 1; with two phosphorothioate internucleotide linkage modifications within positions 1-5 of the sense strand (counting from the 5′-end of the sense strand), 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 of the antisense strand). 
     In one embodiment, n 4  is 0, B3 is 2′-OMe, n 5  is 3, BF is 2′-OMe or 2′-F, q 1  is 9, T1′ is 2′-F, q 2  is 1, B2′ is 2′-OMe or 2′-F, q 3  is 4, T2′ is 2′-F, q 4  is 1, B3′ is 2′-OMe or 2′-F, q 5  is 6, T3′ is 2′-F, q 6  is 1, B4′ is 2′-OMe, and q 7  is 1; with two phosphorothioate internucleotide linkage modifications within positions 1-5 of the sense strand (counting from the 5′-end of the sense strand), 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 of the antisense strand). 
     In one embodiment, B1 is 2′-OMe or 2′-F, n 1  is 8, T1 is 2′F, n 2  is 3, B2 is 2′-OMe, n 3  is 7, n 4  is 0, B3 is 2′OMe, n 5  is 3, BF is 2′-OMe or 2′-F, q 1  is 9, T1′ is 2′-F, q 2  is 1, B2′ is 2′-OMe or 2′- F, q 3  is 4, T2′ is 2′-F, q 4  is 2, B3′ is 2′-OMe or 2′-F, q 5  is 5, T3′ is 2′-F, q 6  is 1, B4′ is 2′-OMe, and q 7  is 1. 
     In one embodiment, B1 is 2′-OMe or 2′-F, n 1  is 8, T1 is 2′F, n 2  is 3, B2 is 2′-OMe, n 3  is 7, n 4  is 0, B3 is 2′-OMe, n 5  is 3, BF is 2′-OMe or 2′-F, q 1  is 9, T1′ is 2′-F, q 2  is 1, B2′ is 2′-OMe or 2′-F, q 3  is 4, T2′ is 2′-F, q 4  is 2, B3′ is 2′-OMe or 2′-F, q 5  is 5, T3′ is 2′-F, q 6  is 1, B4′ is 2′-OMe, and q 7  is 1; with two phosphorothioate internucleotide linkage modifications within positions 1-5 of the sense strand (counting from the 5′-end of the sense strand), 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 of the antisense strand). 
     In one embodiment, B1 is 2′-OMe or 2′-F, n 1  is 6, T1 is 2′F, n 2  is 3, B2 is 2′-OMe, n 3  is 7, n 4  is 0, B3 is 2′OMe, n 5  is 3, BF is 2′-OMe or 2′-F, q 1  is 7, T1′ is 2′-F, q 2  is 1, B2′ is 2′-OMe or 2′- F, q 3  is 4, T2′ is 2′-F, q 4  is 2, B3′ is 2′-OMe or 2′-F, q 5  is 5, T3′ is 2′-F, q 6  is 1, B4′ is 2′-OMe, and q 7  is 1. 
     In one embodiment, B1 is 2′-OMe or 2′-F, n 1  is 6, T1 is 2′F, n 2  is 3, B2 is 2′-OMe, n 3  is 7, n 4  is 0, B3 is 2′-OMe, n 5  is 3, BF is 2′-OMe or 2′-F, q 1  is 7, T1′ is 2′-F, q 2  is 1, B2′ is 2′-OMe or 2′-F, q 3  is 4, T2′ is 2′-F, q 4  is 2, B3′ is 2′-OMe or 2′-F, q 5  is 5, T3′ is 2′-F, q 6  is 1, B4′ is 2′-OMe, and q 7  is 1; with two phosphorothioate internucleotide linkage modifications within positions 1-5 of the sense strand (counting from the 5′-end of the sense strand), 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 of the antisense strand). 
     In one embodiment, B1 is 2′-OMe or 2′-F, n 1  is 8, T1 is 2′F, n 2  is 3, B2 is 2′-OMe, n 3  is 7, n 4  is 0, B3 is 2′OMe, n 5  is 3, BF is 2′-OMe or 2′-F, q 1  is 9, T1′ is 2′-F, q 2  is 1, B2′ is 2′-OMe or 2′- F, q 3  is 4, T2′ is 2′-F, q 4  is 1, B3′ is 2′-OMe or 2′-F, q 5  is 6, T3′ is 2′-F, q 6  is 1, B4′ is 2′-OMe, and q 7  is 1. 
     In one embodiment, B1 is 2′-OMe or 2′-F, n 1  is 8, T1 is 2′F, n 2  is 3, B2 is 2′-OMe, n 3  is 7, n 4  is 0, B3 is 2′-OMe, n 5  is 3, BF is 2′-OMe or 2′-F, q 1  is 9, T1′ is 2′-F, q 2  is 1, B2′ is 2′-OMe or 2′-F, q 3  is 4, T2′ is 2′-F, q 4  is 1, B3′ is 2′-OMe or 2′-F, q 5  is 6, T3′ is 2′-F, q 6  is 1, B4′ is 2′-OMe, and q 7  is 1; with two phosphorothioate internucleotide linkage modifications within positions 1-5 of the sense strand (counting from the 5′-end of the sense strand), 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 of the antisense strand). 
     In one embodiment, B1 is 2′-OMe or 2′-F, n 1  is 8, T1 is 2′F, n 2  is 3, B2 is 2′-OMe, n 3  is 7, n 4  is 0, B3 is 2′OMe, n 5  is 3, BF is 2′-OMe or 2′-F, q 1  is 9, T1′ is 2′-F, q 2  is 1, B2′ is 2′-OMe or 2′- F, q 3  is 5, T2′ is 2′-F, q 4  is 1, B3′ is 2′-OMe or 2′-F, q 5  is 5, T3′ is 2′-F, q 6  is 1, B4′ is 2′-OMe, and q 7  is 1; optionally with at least 2 additional TT at the 3′-end of the antisense strand. 
     In one embodiment, B1 is 2′-OMe or 2′-F, n 1  is 8, T1 is 2′F, n 2  is 3, B2 is 2′-OMe, n 3  is 7, n 4  is 0, B3 is 2′-OMe, n 5  is 3, BF is 2′-OMe or 2′-F, q 1  is 9, T1′ is 2′-F, q 2  is 1, B2′ is 2′-OMe or 2′-F, q 3  is 5, T2′ is 2′-F, q 4  is 1, B3′ is 2′-OMe or 2′-F, q 5  is 5, T3′ is 2′-F, q 6  is 1, B4′ is 2′-OMe, and q 7  is 1; optionally with at least 2 additional TT at the 3′-end of the antisense strand; with two phosphorothioate internucleotide linkage modifications within positions 1-5 of the sense strand (counting from the 5′-end of the sense strand), 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 of the antisense strand). 
     In one embodiment, B1 is 2′-OMe or 2′-F, n 1  is 8, T1 is 2′F, n 2  is 3, B2 is 2′-OMe, n 3  is 7, n 4  is 0, B3 is 2′-OMe, n 5  is 3, BF is 2′-OMe or 2′-F, q 1  is 9, T1′ is 2′-F, q 2  is 1, B2′ is 2′-OMe or 2′- F, q 3  is 4, q 4  is 0, B3′ is 2′-OMe or 2′-F, q 5  is 7, T3′ is 2′-F, q 6  is 1, B4′ is 2′-OMe, and q 7  is 1. In one embodiment, B1 is 2′-OMe or 2′-F, n 1  is 8, T1 is 2′F, n 2  is 3, B2 is 2′-OMe, n 3  is 7, n 4  is 0, B3 is 2′-OMe, n 5  is 3, BF is 2′-OMe or 2′-F, q 1  is 9, T1′ is 2′-F, q 2  is 1, B2′ is 2′-OMe or 2′-F, q 3  is 4, q 4  is 0, B3′ is 2′-OMe or 2′-F, q 5  is 7, T3′ is 2′-F, q 6  is 1, B4′ is 2′-OMe, and q 7  is 1; with two phosphorothioate internucleotide linkage modifications within positions 1-5 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 one embodiment, B1 is 2′-OMe or 2′-F, n 1  is 8, T1 is 2′F, n 2  is 3, B2 is 2′-OMe, n 3  is 7, n 4  is 0, B3 is 2′OMe, n 5  is 3, BF is 2′-OMe or 2′-F, q 1  is 9, T1′ is 2′-F, q 2  is 1, B2′ is 2′-OMe or 2′- F, q 3  is 4, T2′ is 2′-F, q 4  is 2, B3′ is 2′-OMe or 2′-F, q 5  is 5, T3′ is 2′-F, q 6  is 1, B4′ is 2′-F, and q 7  is 1. In one embodiment, B1 is 2′-OMe or 2′-F, n 1  is 8, T1 is 2′F, n 2  is 3, B2 is 2′-OMe, n 3  is 7, n 4  is 0, B3 is 2′-OMe, n 5  is 3, BF is 2′-OMe or 2′-F, q 1  is 9, T1′ is 2′-F, q 2  is 1, B2′ is 2′-OMe or 2′-F, q 3  is 4, T2′ is 2′-F, q 4  is 2, B3′ is 2′-OMe or 2′-F, q 5  is 5, T3′ is 2′-F, q 6  is 1, B4′ is 2′-F, and q 7  is 1; with two phosphorothioate internucleotide linkage modifications within positions 1-5 of the sense strand (counting from the 5′-end of the sense strand), 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 of the antisense strand). 
     In one embodiment, B1 is 2′-OMe or 2′-F, n 1  is 8, T1 is 2′F, n 2  is 3, B2 is 2′-OMe, n 3  is 7, n 4  is 0, B3 is 2′-OMe, n 5  is 3, BF is 2′-OMe or 2′-F, q 1  is 9, T1′ is 2′-F, q 2  is 1, B2′ is 2′-OMe or 2′- F, q 3  is 4, q 4  is 0, B3′ is 2′-OMe or 2′-F, q 5  is 7, T3′ is 2′-F, q 6  is 1, B4′ is 2′-F, and q 7  is 1. In one embodiment, B1 is 2′-OMe or 2′-F, n 1  is 8, T1 is 2′F, n 2  is 3, B2 is 2′-OMe, n 3  is 7, n 4  is 0, B3 is 2′-OMe, n 5  is 3, BF is 2′-OMe or 2′-F, q 1  is 9, T1′ is 2′-F, q 2  is 1, B2′ is 2′-OMe or 2′-F, q 3  is 4, q 4  is 0, B3′ is 2′-OMe or 2′-F, q 5  is 7, T3′ is 2′-F, q 6  is 1, B4′ is 2′-F, and q 7  is 1; with two phosphorothioate internucleotide linkage modifications within positions 1-5 of the sense strand (counting from the 5′-end of the sense strand), 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 of the antisense strand). 
     The RNAi agent can comprise a phosphorus-containing group at the 5′-end of the sense strand or antisense strand. The 5′-end phosphorus-containing group can be 5′-end phosphate (5′-P), 5′-end phosphorothioate (5′-PS), 5′-end phosphorodithioate (5′-PS 2 ), 5′-end vinylphosphonate (5′-VP), 5′-end methylphosphonate (MePhos), or 5′-deoxy-5′-C-malonyl 
     
       
         
         
             
             
         
       
     
     When the 5′-end phosphorus-containing group is 5′-end vinylphosphonate (5′-VP), the 5′-VP can be either 5′-E-VP isomer (i.e., trans-vinyl phosphate, 
     
       
         
         
             
             
         
       
     
     5′-Z-VP isomer (i.e., cis-vinylphosphate, 
     
       
         
         
             
             
         
       
     
     or mixtures thereof. 
     In one embodiment, the RNAi agent comprises a phosphorus-containing group at the 5′-end of the sense strand. In one embodiment, the RNAi agent comprises a phosphorus-containing group at the 5′-end of the antisense strand. 
     In one embodiment, the RNAi agent comprises a 5′-P. In one embodiment, the RNAi agent comprises a 5′-P in the antisense strand. 
     In one embodiment, the RNAi agent comprises a 5′-PS. In one embodiment, the RNAi agent comprises a 5′-PS in the antisense strand. 
     In one embodiment, the RNAi agent comprises a 5′-VP. In one embodiment, the RNAi agent comprises a 5′-VP in the antisense strand. In one embodiment, the RNAi agent comprises a 5′-E-VP in the antisense strand. In one embodiment, the RNAi agent comprises a 5′-Z-VP in the antisense strand. 
     In one embodiment, the RNAi agent comprises a 5′-PS 2 . In one embodiment, the RNAi agent comprises a 5′-PS 2  in the antisense strand. 
     In one embodiment, the RNAi agent comprises a 5′-PS 2 . In one embodiment, the RNAi agent comprises a 5′-deoxy-5′-C-malonyl in the antisense strand. 
     In one embodiment, B1 is 2′-OMe or 2′-F, n 1  is 8, T1 is 2′F, n 2  is 3, B2 is 2′-OMe, n 3  is 7, n 4  is 0, B3 is 2′OMe, n 5  is 3, BF is 2′-OMe or 2′-F, q 1  is 9, T1′ is 2′-F, q 2  is 1, B2′ is 2′-OMe or 2′-F, q 3  is 4, T2′ is 2′-F, q 4  is 2, B3′ is 2′-OMe or 2′-F, q 5  is 5, T3′ is 2′-F, q 6  is 1, B4′ is 2′-OMe, and q 7  is 1. The RNAi agent also comprises a 5′-PS. 
     In one embodiment, B1 is 2′-OMe or 2′-F, n 1  is 8, T1 is 2′F, n 2  is 3, B2 is 2′-OMe, n 3  is 7, n 4  is 0, B3 is 2′OMe, n 5  is 3, BF is 2′-OMe or 2′-F, q 1  is 9, T1′ is 2′-F, q 2  is 1, B2′ is 2′-OMe or 2′- F, q 3  is 4, T2′ is 2′-F, q 4  is 2, B3′ is 2′-OMe or 2′-F, q 5  is 5, T3′ is 2′-F, q 6  is 1, B4′ is 2′-OMe, and q 7  is 1. The RNAi agent also comprises a 5′-P. 
     In one embodiment, B1 is 2′-OMe or 2′-F, n 1  is 8, T1 is 2′F, n 2  is 3, B2 is 2′-OMe, n 3  is 7, n 4  is 0, B3 is 2′OMe, n 5  is 3, BF is 2′-OMe or 2′-F, q 1  is 9, T1′ is 2′-F, q 2  is 1, B2′ is 2′-OMe or 2′- F, q 3  is 4, T2′ is 2′-F, q 4  is 2, B3′ is 2′-OMe or 2′-F, q 5  is 5, T3′ is 2′-F, q 6  is 1, B4′ is 2′-OMe, and q 7  is 1. The RNAi agent also comprises a 5′-VP. The 5′-VP may be 5′-E-VP, 5′-Z-VP, or combination thereof. 
     In one embodiment, B1 is 2′-OMe or 2′-F, n 1  is 8, T1 is 2′F, n 2  is 3, B2 is 2′-OMe, n 3  is 7, n 4  is 0, B3 is 2′OMe, n 5  is 3, BF is 2′-OMe or 2′-F, q 1  is 9, T1′ is 2′-F, q 2  is 1, B2′ is 2′-OMe or 2′- F, q 3  is 4, T2′ is 2′-F, q 4  is 2, B3′ is 2′-OMe or 2′-F, q 5  is 5, T3′ is 2′-F, q 6  is 1, B4′ is 2′-OMe, and q 7  is 1. The RNAi agent also comprises a 5′-PS 2 . 
     In one embodiment, B1 is 2′-OMe or 2′-F, n 1  is 8, T1 is 2′F, n 2  is 3, B2 is 2′-OMe, n 3  is 7, n 4  is 0, B3 is 2′OMe, n 5  is 3, BF is 2′-OMe or 2′-F, q 1  is 9, T1′ is 2′-F, q 2  is 1, B2′ is 2′-OMe or 2′- F, q 3  is 4, T2′ is 2′-F, q 4  is 2, B3′ is 2′-OMe or 2′-F, q 5  is 5, T3′ is 2′-F, q 6  is 1, B4′ is 2′-OMe, and q 7  is 1. The RNAi agent also comprises a 5′-deoxy-5′-C-malonyl. 
     In one embodiment, B1 is 2′-OMe or 2′-F, n 1  is 8, T1 is 2′F, n 2  is 3, B2 is 2′-OMe, n 3  is 7, n 4  is 0, B3 is 2′-OMe, n 5  is 3, BF is 2′-OMe or 2′-F, q 1  is 9, T1′ is 2′-F, q 2  is 1, B2′ is 2′-OMe or 2′- F, q 3  is 4, T2′ is 2′-F, q 4  is 2, B3′ is 2′-OMe or 2′-F, q 5  is 5, T3′ is 2′-F, q 6  is 1, B4′ is 2′-OMe, and q 7  is 1; with two phosphorothioate internucleotide linkage modifications within position 1-5 of the sense strand (counting from the 5′-end of the sense strand), 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 of the antisense strand). The RNAi agent also comprises a 5′-P. 
     In one embodiment, B1 is 2′-OMe or 2′-F, n 1  is 8, T1 is 2′F, n 2  is 3, B2 is 2′-OMe, n 3  is 7, n 4  is 0, B3 is 2′-OMe, n 5  is 3, BF is 2′-OMe or 2′-F, q 1  is 9, T1′ is 2′-F, q 2  is 1, B2′ is 2′-OMe or 2′- F, q 3  is 4, T2′ is 2′-F, q 4  is 2, B3′ is 2′-OMe or 2′-F, q 5  is 5, T3′ is 2′-F, q 6  is 1, B4′ is 2′-OMe, and q 7  is 1; with two phosphorothioate internucleotide linkage modifications within position 1-5 of the sense strand (counting from the 5′-end of the sense strand), 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 of the antisense strand). The RNAi agent also comprises a 5′-PS. 
     In one embodiment, B1 is 2′-OMe or 2′-F, n 1  is 8, T1 is 2′F, n 2  is 3, B2 is 2′-OMe, n 3  is 7, n 4  is 0, B3 is 2′-OMe, n 5  is 3, BF is 2′-OMe or 2′-F, q 1  is 9, T1′ is 2′-F, q 2  is 1, B2′ is 2′-OMe or 2′- F, q 3  is 4, T2′ is 2′-F, q 4  is 2, B3′ is 2′-OMe or 2′-F, q 5  is 5, T3′ is 2′-F, q 6  is 1, B4′ is 2′-OMe, and q 7  is 1; with two phosphorothioate internucleotide linkage modifications within position 1-5 of the sense strand (counting from the 5′-end of the sense strand), 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 of the antisense strand). The RNAi agent also comprises a 5′-VP. The 5′-VP may be 5′-E-VP, 5′-Z-VP, or combination thereof. 
     In one embodiment, B1 is 2′-OMe or 2′-F, n 1  is 8, T1 is 2′F, n 2  is 3, B2 is 2′-OMe, n 3  is 7, n 4  is 0, B3 is 2′-OMe, n 5  is 3, BF is 2′-OMe or 2′-F, q 1  is 9, T1′ is 2′-F, q 2  is 1, B2′ is 2′-OMe or 2′-F, q 3  is 4, T2′ is 2′-F, q 4  is 2, B3′ is 2′-OMe or 2′-F, q 5  is 5, T3′ is 2′-F, q 6  is 1, B4′ is 2′-OMe, and q 7  is 1; with two phosphorothioate internucleotide linkage modifications within position 1-5 of the sense strand (counting from the 5′-end of the sense strand), 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 of the antisense strand). The RNAi agent also comprises a 5′-PS 2 . 
     In one embodiment, B1 is 2′-OMe or 2′-F, n 1  is 8, T1 is 2′F, n 2  is 3, B2 is 2′-OMe, n 3  is 7, n 4  is 0, B3 is 2′-OMe, n 5  is 3, BF is 2′-OMe or 2′-F, q 1  is 9, T1′ is 2′-F, q 2  is 1, B2′ is 2′-OMe or 2′- F, q 3  is 4, T2′ is 2′-F, q 4  is 2, B3′ is 2′-OMe or 2′-F, q 5  is 5, T3′ is 2′-F, q 6  is 1, B4′ is 2′-OMe, and q 7  is 1; with two phosphorothioate internucleotide linkage modifications within position 1-5 of the sense strand (counting from the 5′-end of the sense strand), 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 of the antisense strand). The RNAi agent also comprises a 5′-deoxy-5′-C-malonyl. 
     In one embodiment, B1 is 2′-OMe or 2′-F, n 1  is 8, T1 is 2′F, n 2  is 3, B2 is 2′-OMe, n 3  is 7, n 4  is 0, B3 is 2′-OMe, n 5  is 3, BF is 2′-OMe or 2′-F, q 1  is 9, T1′ is 2′-F, q 2  is 1, B2′ is 2′-OMe or 2′- F, q 3  is 4, q 4  is 0, B3′ is 2′-OMe or 2′-F, q 5  is 7, T3′ is 2′-F, q 6  is 1, B4′ is 2′-OMe, and q 7  is 1. The RNAi agent also comprises a 5′-P. 
     In one embodiment, B1 is 2′-OMe or 2′-F, n 1  is 8, T1 is 2′F, n 2  is 3, B2 is 2′-OMe, n 3  is 7, n 4  is 0, B3 is 2′-OMe, n 5  is 3, BF is 2′-OMe or 2′-F, q 1  is 9, T1′ is 2′-F, q 2  is 1, B2′ is 2′-OMe or 2′- F, q 3  is 4, q 4  is 0, B3′ is 2′-OMe or 2′-F, q 5  is 7, T3′ is 2′-F, q 6  is 1, B4′ is 2′-OMe, and q 7  is 1. The dsRNA agent also comprises a 5′-PS. 
     In one embodiment, B1 is 2′-OMe or 2′-F, n 1  is 8, T1 is 2′F, n 2  is 3, B2 is 2′-OMe, n 3  is 7, n 4  is 0, B3 is 2′-OMe, n 5  is 3, BF is 2′-OMe or 2′-F, q 1  is 9, T1′ is 2′-F, q 2  is 1, B2′ is 2′-OMe or 2′- F, q 3  is 4, q 4  is 0, B3′ is 2′-OMe or 2′-F, q 5  is 7, T3′ is 2′-F, q 6  is 1, B4′ is 2′-OMe, and q 7  is 1. The RNAi agent also comprises a 5′-VP. The 5′-VP may be 5′-E-VP, 5′-Z-VP, or combination thereof. In one embodiment, B1 is 2′-OMe or 2′-F, n 1  is 8, T1 is 2′F, n 2  is 3, B2 is 2′-OMe, n 3  is 7, n 4  is 0, B3 is 2′-OMe, n 5  is 3, BF is 2′-OMe or 2′-F, q 1  is 9, T1′ is 2′-F, q 2  is 1, B2′ is 2′-OMe or 2′-F, q 3  is 4, q 4  is 0, B3′ is 2′-OMe or 2′-F, q 5  is 7, T3′ is 2′-F, q 6  is 1, B4′ is 2′-OMe, and q 7  is 1. The RNAi agent also comprises a 5′-PS 2 . 
     In one embodiment, B1 is 2′-OMe or 2′-F, n 1  is 8, T1 is 2′F, n 2  is 3, B2 is 2′-OMe, n 3  is 7, n 4  is 0, B3 is 2′-OMe, n 5  is 3, BF is 2′-OMe or 2′-F, q 1  is 9, T1′ is 2′-F, q 2  is 1, B2′ is 2′-OMe or 2′- F, q 3  is 4, q 4  is 0, B3′ is 2′-OMe or 2′-F, q 5  is 7, T3′ is 2′-F, q 6  is 1, B4′ is 2′-OMe, and q 7  is 1. The RNAi agent also comprises a 5′-deoxy-5′-C-malonyl. 
     In one embodiment, B1 is 2′-OMe or 2′-F, n 1  is 8, T1 is 2′F, n 2  is 3, B2 is 2′-OMe, n 3  is 7, n 4  is 0, B3 is 2′-OMe, n 5  is 3, BF is 2′-OMe or 2′-F, q 1  is 9, T1′ is 2′-F, q 2  is 1, B2′ is 2′-OMe or 2′- F, q 3  is 4, q 4  is 0, B3′ is 2′-OMe or 2′-F, q 5  is 7, T3′ is 2′-F, q 6  is 1, B4′ is 2′-OMe, and q 7  is 1; with two phosphorothioate internucleotide linkage modifications within position 1-5 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). The RNAi agent also comprises a 5′-P. 
     In one embodiment, B1 is 2′-OMe or 2′-F, n 1  is 8, T1 is 2′F, n 2  is 3, B2 is 2′-OMe, n 3  is 7, n 4  is 0, B3 is 2′-OMe, n 5  is 3, BF is 2′-OMe or 2′-F, q 1  is 9, T1′ is 2′-F, q 2  is 1, B2′ is 2′-OMe or 2′- F, q 3  is 4, q 4  is 0, B3′ is 2′-OMe or 2′-F, q 5  is 7, T3′ is 2′-F, q 6  is 1, B4′ is 2′-OMe, and q 7  is 1; with two phosphorothioate internucleotide linkage modifications within position 1-5 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). The RNAi agent also comprises a 5′-PS. 
     In one embodiment, B1 is 2′-OMe or 2′-F, n 1  is 8, T1 is 2′F, n 2  is 3, B2 is 2′-OMe, n 3  is 7, n 4  is 0, B3 is 2′-OMe, n 5  is 3, BF is 2′-OMe or 2′-F, q 1  is 9, T1′ is 2′-F, q 2  is 1, B2′ is 2′-OMe or 2′- F, q 3  is 4, q 4  is 0, B3′ is 2′-OMe or 2′-F, q 5  is 7, T3′ is 2′-F, q 6  is 1, B4′ is 2′-OMe, and q 7  is 1; with two phosphorothioate internucleotide linkage modifications within position 1-5 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). The RNAi agent also comprises a 5′-VP. The 5′-VP may be 5′-E-VP, 5′-Z-VP, or combination thereof. 
     In one embodiment, B1 is 2′-OMe or 2′-F, n 1  is 8, T1 is 2′F, n 2  is 3, B2 is 2′-OMe, n 3  is 7, n 4  is 0, B3 is 2′-OMe, n 5  is 3, BF is 2′-OMe or 2′-F, q 1  is 9, T1′ is 2′-F, q 2  is 1, B2′ is 2′-OMe or 2′- F, q 3  is 4, q 4  is 0, B3′ is 2′-OMe or 2′-F, q 5  is 7, T3′ is 2′-F, q 6  is 1, B4′ is 2′-OMe, and q 7  is 1; with two phosphorothioate internucleotide linkage modifications within position 1-5 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). The RNAi agent also comprises a 5′-PS 2 . 
     In one embodiment, B1 is 2′-OMe or 2′-F, n 1  is 8, T1 is 2′F, n 2  is 3, B2 is 2′-OMe, n 3  is 7, n 4  is 0, B3 is 2′-OMe, n 5  is 3, BF is 2′-OMe or 2′-F, q 1  is 9, T1′ is 2′-F, q 2  is 1, B2′ is 2′-OMe or 2′- F, q 3  is 4, q 4  is 0, B3′ is 2′-OMe or 2′-F, q 5  is 7, T3′ is 2′-F, q 6  is 1, B4′ is 2′-OMe, and q 7  is 1; with two phosphorothioate internucleotide linkage modifications within position 1-5 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). The RNAi agent also comprises a 5′-deoxy-5′-C-malonyl. 
     In one embodiment, B1 is 2′-OMe or 2′-F, n 1  is 8, T1 is 2′F, n 2  is 3, B2 is 2′-OMe, n 3  is 7, n 4  is 0, B3 is 2′OMe, n 5  is 3, BF is 2′-OMe or 2′-F, q 1  is 9, T1′ is 2′-F, q 2  is 1, B2′ is 2′-OMe or 2′-F, q 3  is 4, T2′ is 2′-F, q 4  is 2, B3′ is 2′-OMe or 2′-F, q 5  is 5, T3′ is 2′-F, q 6  is 1, B4′ is 2′-F, and q 7  is 1. The RNAi agent also comprises a 5′-P. 
     In one embodiment, B1 is 2′-OMe or 2′-F, n 1  is 8, T1 is 2′F, n 2  is 3, B2 is 2′-OMe, n 3  is 7, n 4  is 0, B3 is 2′OMe, n 5  is 3, BF is 2′-OMe or 2′-F, q 1  is 9, T1′ is 2′-F, q 2  is 1, B2′ is 2′-OMe or 2′- F, q 3  is 4, T2′ is 2′-F, q 4  is 2, B3′ is 2′-OMe or 2′-F, q 5  is 5, T3′ is 2′-F, q 6  is 1, B4′ is 2′-F, and q 7  is 1. The RNAi agent also comprises a 5′-PS. 
     In one embodiment, B1 is 2′-OMe or 2′-F, n 1  is 8, T1 is 2′F, n 2  is 3, B2 is 2′-OMe, n 3  is 7, n 4  is 0, B3 is 2′OMe, n 5  is 3, BF is 2′-OMe or 2′-F, q 1  is 9, T1′ is 2′-F, q 2  is 1, B2′ is 2′-OMe or 2′- F, q 3  is 4, T2′ is 2′-F, q 4  is 2, B3′ is 2′-OMe or 2′-F, q 5  is 5, T3′ is 2′-F, q 6  is 1, B4′ is 2′-F, and q 7  is 1. The RNAi agent also comprises a 5′-VP. The 5′-VP may be 5′-E-VP, 5′-Z-VP, or combination thereof. 
     In one embodiment, B1 is 2′-OMe or 2′-F, n 1  is 8, T1 is 2′F, n 2  is 3, B2 is 2′-OMe, n 3  is 7, n 4  is 0, B3 is 2′OMe, n 5  is 3, BF is 2′-OMe or 2′-F, q 1  is 9, T1′ is 2′-F, q 2  is 1, B2′ is 2′-OMe or 2′-F, q 3  is 4, T2′ is 2′-F, q 4  is 2, B3′ is 2′-OMe or 2′-F, q 5  is 5, T3′ is 2′-F, q 6  is 1, B4′ is 2′-F, and q 7  is 1. The dsRNAi RNA agent also comprises a 5′-PS 2 . 
     In one embodiment, B1 is 2′-OMe or 2′-F, n 1  is 8, T1 is 2′F, n 2  is 3, B2 is 2′-OMe, n 3  is 7, n 4  is 0, B3 is 2′OMe, n 5  is 3, BF is 2′-OMe or 2′-F, q 1  is 9, T1′ is 2′-F, q 2  is 1, B2′ is 2′-OMe or 2′- F, q 3  is 4, T2′ is 2′-F, q 4  is 2, B3′ is 2′-OMe or 2′-F, q 5  is 5, T3′ is 2′-F, q 6  is 1, B4′ is 2′-F, and q 7  is 1. The RNAi agent also comprises a 5′-deoxy-5′-C-malonyl. 
     In one embodiment, B1 is 2′-OMe or 2′-F, n 1  is 8, T1 is 2′F, n 2  is 3, B2 is 2′-OMe, n 3  is 7, n 4  is 0, B3 is 2′-OMe, n 5  is 3, BF is 2′-OMe or 2′-F, q 1  is 9, T1′ is 2′-F, q 2  is 1, B2′ is 2′-OMe or 2′- F, q 3  is 4, T2′ is 2′-F, q 4  is 2, B3′ is 2′-OMe or 2′-F, q 5  is 5, T3′ is 2′-F, q 6  is 1, B4′ is 2′-F, and q 7  is 1; with two phosphorothioate internucleotide linkage modifications within position 1-5 of the sense strand (counting from the 5′-end of the sense strand), 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 of the antisense strand). The RNAi agent also comprises a 5′-P. 
     In one embodiment, B1 is 2′-OMe or 2′-F, n 1  is 8, T1 is 2′F, n 2  is 3, B2 is 2′-OMe, n 3  is 7, n 4  is 0, B3 is 2′-OMe, n 5  is 3, BF is 2′-OMe or 2′-F, q 1  is 9, T1′ is 2′-F, q 2  is 1, B2′ is 2′-OMe or 2′- F, q 3  is 4, T2′ is 2′-F, q 4  is 2, B3′ is 2′-OMe or 2′-F, q 5  is 5, T3′ is 2′-F, q 6  is 1, B4′ is 2′-F, and q 7  is 1; with two phosphorothioate internucleotide linkage modifications within position 1-5 of the sense strand (counting from the 5′-end of the sense strand), 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 of the antisense strand). The RNAi agent also comprises a 5′-PS. 
     In one embodiment, B1 is 2′-OMe or 2′-F, n 1  is 8, T1 is 2′F, n 2  is 3, B2 is 2′-OMe, n 3  is 7, n 4  is 0, B3 is 2′-OMe, n 5  is 3, BF is 2′-OMe or 2′-F, q 1  is 9, T1′ is 2′-F, q 2  is 1, B2′ is 2′-OMe or 2′- F, q 3  is 4, T2′ is 2′-F, q 4  is 2, B3′ is 2′-OMe or 2′-F, q 5  is 5, T3′ is 2′-F, q 6  is 1, B4′ is 2′-F, and q 7  is 1; with two phosphorothioate internucleotide linkage modifications within position 1-5 of the sense strand (counting from the 5′-end of the sense strand), 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 of the antisense strand). The RNAi agent also comprises a 5′-VP. The 5′-VP may be 5′-E-VP, 5′-Z-VP, or combination thereof. 
     In one embodiment, B1 is 2′-OMe or 2′-F, n 1  is 8, T1 is 2′F, n 2  is 3, B2 is 2′-OMe, n 3  is 7, n 4  is 0, B3 is 2′-OMe, n 5  is 3, BF is 2′-OMe or 2′-F, q 1  is 9, T1′ is 2′-F, q 2  is 1, B2′ is 2′-OMe or 2′- F, q 3  is 4, T2′ is 2′-F, q 4  is 2, B3′ is 2′-OMe or 2′-F, q 5  is 5, T3′ is 2′-F, q 6  is 1, B4′ is 2′-F, and q 7  is 1; with two phosphorothioate internucleotide linkage modifications within position 1-5 of the sense strand (counting from the 5′-end of the sense strand), 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 of the antisense strand). The RNAi agent also comprises a 5′-PS 2 . 
     In one embodiment, B1 is 2′-OMe or 2′-F, n 1  is 8, T1 is 2′F, n 2  is 3, B2 is 2′-OMe, n 3  is 7, n 4  is 0, B3 is 2′-OMe, n 5  is 3, BF is 2′-OMe or 2′-F, q 1  is 9, T1′ is 2′-F, q 2  is 1, B2′ is 2′-OMe or 2′- F, q 3  is 4, T2′ is 2′-F, q 4  is 2, B3′ is 2′-OMe or 2′-F, q 5  is 5, T3′ is 2′-F, q 6  is 1, B4′ is 2′-F, and q 7  is 1; with two phosphorothioate internucleotide linkage modifications within position 1-5 of the sense strand (counting from the 5′-end of the sense strand), 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 of the antisense strand). The RNAi agent also comprises a 5′-deoxy-5′-C-malonyl. 
     In one embodiment, B1 is 2′-OMe or 2′-F, n 1  is 8, T1 is 2′F, n 2  is 3, B2 is 2′-OMe, n 3  is 7, n 4  is 0, B3 is 2′-OMe, n 5  is 3, BF is 2′-OMe or 2′-F, q 1  is 9, T1′ is 2′-F, q 2  is 1, B2′ is 2′-OMe or 2′- F, q 3  is 4, q 4  is 0, B3′ is 2′-OMe or 2′-F, q 5  is 7, T3′ is 2′-F, q 6  is 1, B4′ is 2′-F, and q 7  is 1. The RNAi agent also comprises a 5′-P. 
     In one embodiment, B1 is 2′-OMe or 2′-F, n 1  is 8, T1 is 2′F, n 2  is 3, B2 is 2′-OMe, n 3  is 7, n 4  is 0, B3 is 2′-OMe, n 5  is 3, BF is 2′-OMe or 2′-F, q 1  is 9, T1′ is 2′-F, q 2  is 1, B2′ is 2′-OMe or 2′- F, q 3  is 4, q 4  is 0, B3′ is 2′-OMe or 2′-F, q 5  is 7, T3′ is 2′-F, q 6  is 1, B4′ is 2′-F, and q 7  is 1. The RNAi agent also comprises a 5′-PS. 
     In one embodiment, B1 is 2′-OMe or 2′-F, n 1  is 8, T1 is 2′F, n 2  is 3, B2 is 2′-OMe, n 3  is 7, n 4  is 0, B3 is 2′-OMe, n 5  is 3, BF is 2′-OMe or 2′-F, q 1  is 9, T1′ is 2′-F, q 2  is 1, B2′ is 2′-OMe or 2′- F, q 3  is 4, q 4  is 0, B3′ is 2′-OMe or 2′-F, q 5  is 7, T3′ is 2′-F, q 6  is 1, B4′ is 2′-F, and q 7  is 1. The RNAi agent also comprises a 5′-VP. The 5′-VP may be 5′-E-VP, 5′-Z-VP, or combination thereof. 
     In one embodiment, B1 is 2′-OMe or 2′-F, n 1  is 8, T1 is 2′F, n 2  is 3, B2 is 2′-OMe, n 3  is 7, n 4  is 0, B3 is 2′-OMe, n 5  is 3, BF is 2′-OMe or 2′-F, q 1  is 9, T1′ is 2′-F, q 2  is 1, B2′ is 2′-OMe or 2′- F, q 3  is 4, q 4  is 0, B3′ is 2′-OMe or 2′-F, q 5  is 7, T3′ is 2′-F, q 6  is 1, B4′ is 2′-F, and q 7  is 1. The RNAi agent also comprises a 5′-PS 2 . 
     In one embodiment, B1 is 2′-OMe or 2′-F, n 1  is 8, T1 is 2′F, n 2  is 3, B2 is 2′-OMe, n 3  is 7, n 4  is 0, B3 is 2′-OMe, n 5  is 3, BF is 2′-OMe or 2′-F, q 1  is 9, T1′ is 2′-F, q 2  is 1, B2′ is 2′-OMe or 2′- F, q 3  is 4, q 4  is 0, B3′ is 2′-OMe or 2′-F, q 5  is 7, T3′ is 2′-F, q 6  is 1, B4′ is 2′-F, and q 7  is 1. The RNAi agent also comprises a 5′-deoxy-5′-C-malonyl. 
     In one embodiment, B1 is 2′-OMe or 2′-F, n 1  is 8, T1 is 2′F, n 2  is 3, B2 is 2′-OMe, n 3  is 7, n 4  is 0, B3 is 2′-OMe, n 5  is 3, BF is 2′-OMe or 2′-F, q 1  is 9, T1′ is 2′-F, q 2  is 1, B2′ is 2′-OMe or 2′- F, q 3  is 4, q 4  is 0, B3′ is 2′-OMe or 2′-F, q 5  is 7, T3′ is 2′-F, q 6  is 1, B4′ is 2′-F, and q 7  is 1; with two phosphorothioate internucleotide linkage modifications within position 1-5 of the sense strand (counting from the 5′-end of the sense strand), 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 of the antisense strand). The RNAi agent also comprises a 5′-P. 
     In one embodiment, B1 is 2′-OMe or 2′-F, n 1  is 8, T1 is 2′F, n 2  is 3, B2 is 2′-OMe, n 3  is 7, n 4  is 0, B3 is 2′-OMe, n 5  is 3, BF is 2′-OMe or 2′-F, q 1  is 9, T1′ is 2′-F, q 2  is 1, B2′ is 2′-OMe or 2′- F, q 3  is 4, q 4  is 0, B3′ is 2′-OMe or 2′-F, q 5  is 7, T3′ is 2′-F, q 6  is 1, B4′ is 2′-F, and q 7  is 1; with two phosphorothioate internucleotide linkage modifications within position 1-5 of the sense strand (counting from the 5′-end of the sense strand), 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 of the antisense strand). The RNAi agent also comprises a 5′-PS. 
     In one embodiment, B1 is 2′-OMe or 2′-F, n 1  is 8, T1 is 2′F, n 2  is 3, B2 is 2′-OMe, n 3  is 7, n 4  is 0, B3 is 2′-OMe, n 5  is 3, BF is 2′-OMe or 2′-F, q 1  is 9, T1′ is 2′-F, q 2  is 1, B2′ is 2′-OMe or 2′- F, q 3  is 4, q 4  is 0, B3′ is 2′-OMe or 2′-F, q 5  is 7, T3′ is 2′-F, q 6  is 1, B4′ is 2′-F, and q 7  is 1; with two phosphorothioate internucleotide linkage modifications within position 1-5 of the sense strand (counting from the 5′-end of the sense strand), 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 of the antisense strand). The RNAi agent also comprises a 5′-VP. The 5′-VP may be 5′-E-VP, 5′-Z-VP, or combination thereof. 
     In one embodiment, B1 is 2′-OMe or 2′-F, n 1  is 8, T1 is 2′F, n 2  is 3, B2 is 2′-OMe, n 3  is 7, n 4  is 0, B3 is 2′-OMe, n 5  is 3, BF is 2′-OMe or 2′-F, q 1  is 9, T1′ is 2′-F, q 2  is 1, B2′ is 2′-OMe or 2′- F, q 3  is 4, q 4  is 0, B3′ is 2′-OMe or 2′-F, q 5  is 7, T3′ is 2′-F, q 6  is 1, B4′ is 2′-F, and q 7  is 1; with two phosphorothioate internucleotide linkage modifications within position 1-5 of the sense strand (counting from the 5′-end of the sense strand), 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 of the antisense strand). The RNAi agent also comprises a 5′-PS 2 . 
     In one embodiment, B1 is 2′-OMe or 2′-F, n 1  is 8, T1 is 2′F, n 2  is 3, B2 is 2′-OMe, n 3  is 7, n 4  is 0, B3 is 2′-OMe, n 5  is 3, B1′ is 2′-OMe or 2′-F, q 1  is 9, T1′ is 2′-F, q 2  is 1, B2′ is 2′-OMe or 2′-F, q 3  is 4, q 4  is 0, B3′ is 2′-OMe or 2′-F, q 5  is 7, T3′ is 2′-F, q 6  is 1, B4′ is 2′-F, and q 7  is 1; with two phosphorothioate internucleotide linkage modifications within position 1-5 of the sense strand (counting from the 5′-end of the sense strand), 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 of the antisense strand). The RNAi agent also comprises a 5′-deoxy-5′-C-malonyl. 
     In one embodiment, B1 is 2′-OMe or 2′-F, n 1  is 8, T1 is 2′F, n 2  is 3, B2 is 2′-OMe, n 3  is 7, n 4  is 0, B3 is 2′-OMe, n 5  is 3, B1′ is 2′-OMe or 2′-F, q 1  is 9, T1′ is 2′-F, q 2  is 1, B2′ is 2′-OMe or 2′-F, q 3  is 4, T2′ is 2′-F, q 4  is 2, B3′ is 2′-OMe or 2′-F, q 5  is 5, T3′ is 2′-F, q 6  is 1, B4′ is 2′-OMe, and q 7  is 1; with two phosphorothioate internucleotide linkage modifications within position 1-5 of the sense strand (counting from the 5′-end of the sense strand), 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 of the antisense strand). The RNAi agent also comprises a 5′-P and a targeting ligand. In one embodiment, the 5′-P is at the 5′-end of the antisense strand, and the targeting ligand is at the 3′-end of the sense strand. 
     In one embodiment, B1 is 2′-OMe or 2′-F, n 1  is 8, T1 is 2′F, n 2  is 3, B2 is 2′-OMe, n 3  is 7, n 4  is 0, B3 is 2′-OMe, n 5  is 3, B1′ is 2′-OMe or 2′-F, q 1  is 9, T1′ is 2′-F, q 2  is 1, B2′ is 2′-OMe or 2′-F, q 3  is 4, T2′ is 2′-F, q 4  is 2, B3′ is 2′-OMe or 2′-F, q 5  is 5, T3′ is 2′-F, q 6  is 1, B4′ is 2′-OMe, and q 7  is 1; with two phosphorothioate internucleotide linkage modifications within position 1-5 of the sense strand (counting from the 5′-end of the sense strand), 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 of the antisense strand). The RNAi agent also comprises a 5′-PS and a targeting ligand. In one embodiment, the 5′-PS is at the 5′-end of the antisense strand, and the targeting ligand is at the 3′-end of the sense strand. 
     In one embodiment, B1 is 2′-OMe or 2′-F, n 1  is 8, T1 is 2′F, n 2  is 3, B2 is 2′-OMe, n 3  is 7, n 4  is 0, B3 is 2′-OMe, n 5  is 3, B1′ is 2′-OMe or 2′-F, q 1  is 9, T1′ is 2′-F, q 2  is 1, B2′ is 2′-OMe or 2′-F, q 3  is 4, T2′ is 2′-F, q 4  is 2, B3′ is 2′-OMe or 2′-F, q 5  is 5, T3′ is 2′-F, q 6  is 1, B4′ is 2′-OMe, and q 7  is 1; with two phosphorothioate internucleotide linkage modifications within position 1-5 of the sense strand (counting from the 5′-end of the sense strand), 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 of the antisense strand). The RNAi agent also comprises a 5′-VP (e.g., a 5′-E-VP, 5′-Z-VP, or combination thereof), and a targeting ligand. 
     In one embodiment, the 5′-VP is at the 5′-end of the antisense strand, and the targeting ligand is at the 3′-end of the sense strand. 
     In one embodiment, B1 is 2′-OMe or 2′-F, n 1  is 8, T1 is 2′F, n 2  is 3, B2 is 2′-OMe, n 3  is 7, n 4  is 0, B3 is 2′-OMe, n 5  is 3, B1′ is 2′-OMe or 2′-F, q 1  is 9, T1′ is 2′-F, q 2  is 1, B2′ is 2′-OMe or 2′-F, q 3  is 4, T2′ is 2′-F, q 4  is 2, B3′ is 2′-OMe or 2′-F, q 5  is 5, T3′ is 2′-F, q 6  is 1, B4′ is 2′-OMe, and q 7  is 1; with two phosphorothioate internucleotide linkage modifications within position 1-5 of the sense strand (counting from the 5′-end of the sense strand), 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 of the antisense strand). The RNAi agent also comprises a 5′-PS 2  and a targeting ligand. In one embodiment, the 5′-PS 2  is at the 5′-end of the antisense strand, and the targeting ligand is at the 3′-end of the sense strand. 
     In one embodiment, B1 is 2′-OMe or 2′-F, n 1  is 8, T1 is 2′F, n 2  is 3, B2 is 2′-OMe, n 3  is 7, n 4  is 0, B3 is 2′-OMe, n 5  is 3, B1′ is 2′-OMe or 2′-F, q 1  is 9, T1′ is 2′-F, q 2  is 1, B2′ is 2′-OMe or 2′-F, q 3  is 4, T2′ is 2′-F, q 4  is 2, B3′ is 2′-OMe or 2′-F, q 5  is 5, T3′ is 2′-F, q 6  is 1, B4′ is 2′-OMe, and q 7  is 1; with two phosphorothioate internucleotide linkage modifications within position 1-5 of the sense strand (counting from the 5′-end of the sense strand), 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 of the antisense strand). The RNAi agent also comprises a 5′-deoxy-5′-C-malonyl and a targeting ligand. In one embodiment, the 5′-deoxy-5′-C-malonyl is at the 5′-end of the antisense strand, and the targeting ligand is at the 3′-end of the sense strand. 
     In one embodiment, B1 is 2′-OMe or 2′-F, n 1  is 8, T1 is 2′F, n 2  is 3, B2 is 2′-OMe, n 3  is 7, n 4  is 0, B3 is 2′-OMe, n 5  is 3, B1′ is 2′-OMe or 2′-F, q 1  is 9, T1′ is 2′-F, q 2  is 1, B2′ is 2′-OMe or 2′-F, q 3  is 4, q 4  is 0, B3′ is 2′-OMe or 2′-F, q 5  is 7, T3′ is 2′-F, q 6  is 1, B4′ is 2′-OMe, and q 7  is 1; with two phosphorothioate internucleotide linkage modifications within position 1-5 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). The RNAi agent also comprises a 5′-P and a targeting ligand. In one embodiment, the 5′-P is at the 5′-end of the antisense strand, and the targeting ligand is at the 3′-end of the sense strand. 
     In one embodiment, B1 is 2′-OMe or 2′-F, n 1  is 8, T1 is 2′F, n 2  is 3, B2 is 2′-OMe, n 3  is 7, n 4  is 0, B3 is 2′-OMe, n 5  is 3, B1′ is 2′-OMe or 2′-F, q 1  is 9, T1′ is 2′-F, q 2  is 1, B2′ is 2′-OMe or 2′-F, q 3  is 4, q 4  is 0, B3′ is 2′-OMe or 2′-F, q 5  is 7, T3′ is 2′-F, q 6  is 1, B4′ is 2′-OMe, and q 7  is 1; with two phosphorothioate internucleotide linkage modifications within position 1-5 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). The RNAi agent also comprises a 5′-PS and a targeting ligand. In one embodiment, the 5′-PS is at the 5′-end of the antisense strand, and the targeting ligand is at the 3′-end of the sense strand. 
     In one embodiment, B1 is 2′-OMe or 2′-F, n 1  is 8, T1 is 2′F, n 2  is 3, B2 is 2′-OMe, n 3  is 7, n 4  is 0, B3 is 2′-OMe, n 5  is 3, BF is 2′-OMe or 2′-F, q 1  is 9, T1′ is 2′-F, q 2  is 1, B2′ is 2′-OMe or 2′- F, q 3  is 4, q 4  is 0, B3′ is 2′-OMe or 2′-F, q 5  is 7, T3′ is 2′-F, q 6  is 1, B4′ is 2′-OMe, and q 7  is 1; with two phosphorothioate internucleotide linkage modifications within position 1-5 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). The RNAi agent also comprises a 5′-VP (e.g., a 5′-E-VP, 5′-Z-VP, or combination thereof) and a targeting ligand. In one embodiment, the 5′-VP is at the 5′-end of the antisense strand, and the targeting ligand is at the 3′-end of the sense strand. 
     In one embodiment, B1 is 2′-OMe or 2′-F, n 1  is 8, T1 is 2′F, n 2  is 3, B2 is 2′-OMe, n 3  is 7, n 4  is 0, B3 is 2′-OMe, n 5  is 3, BF is 2′-OMe or 2′-F, q 1  is 9, T1′ is 2′-F, q 2  is 1, B2′ is 2′-OMe or 2′- F, q 3  is 4, q 4  is 0, B3′ is 2′-OMe or 2′-F, q 5  is 7, T3′ is 2′-F, q 6  is 1, B4′ is 2′-OMe, and q 7  is 1; with two phosphorothioate internucleotide linkage modifications within position 1-5 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). The RNAi agent also comprises a 5′-PS 2  and a targeting ligand. In one embodiment, the 5′-PS 2  is at the 5′-end of the antisense strand, and the targeting ligand is at the 3′-end of the sense strand. 
     In one embodiment, B1 is 2′-OMe or 2′-F, n 1  is 8, T1 is 2′F, n 2  is 3, B2 is 2′-OMe, n 3  is 7, n 4  is 0, B3 is 2′-OMe, n 5  is 3, BF is 2′-OMe or 2′-F, q 1  is 9, T1′ is 2′-F, q 2  is 1, B2′ is 2′-OMe or 2′- F, q 3  is 4, q 4  is 0, B3′ is 2′-OMe or 2′-F, q 5  is 7, T3′ is 2′-F, q 6  is 1, B4′ is 2′-OMe, and q 7  is 1; with two phosphorothioate internucleotide linkage modifications within position 1-5 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). The RNAi agent also comprises a 5′-deoxy-5′-C-malonyl and a targeting ligand. In one embodiment, the 5′-deoxy-5′-C-malonyl is at the 5′-end of the antisense strand, and the targeting ligand is at the 3′-end of the sense strand. 
     In one embodiment, B1 is 2′-OMe or 2′-F, n 1  is 8, T1 is 2′F, n 2  is 3, B2 is 2′-OMe, n 3  is 7, n 4  is 0, B3 is 2′-OMe, n 5  is 3, BF is 2′-OMe or 2′-F, q 1  is 9, T1′ is 2′-F, q 2  is 1, B2′ is 2′-OMe or 2′- F, q 3  is 4, T2′ is 2′-F, q 4  is 2, B3′ is 2′-OMe or 2′-F, q 5  is 5, T3′ is 2′-F, q 6  is 1, B4′ is 2′-F, and q 7  is 1; with two phosphorothioate internucleotide linkage modifications within position 1-5 of the sense strand (counting from the 5′-end of the sense strand), 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 of the antisense strand). The RNAi agent also comprises a 5′-P and a targeting ligand. In one embodiment, the 5′-P is at the 5′-end of the antisense strand, and the targeting ligand is at the 3′-end of the sense strand. 
     In one embodiment, B1 is 2′-OMe or 2′-F, n 1  is 8, T1 is 2′F, n 2  is 3, B2 is 2′-OMe, n 3  is 7, n 4  is 0, B3 is 2′-OMe, n 5  is 3, B1′ is 2′-OMe or 2′-F, q 1  is 9, T1′ is 2′-F, q 2  is 1, B2′ is 2′-OMe or 2′-F, q 3  is 4, T2′ is 2′-F, q 4  is 2, B3′ is 2′-OMe or 2′-F, q 5  is 5, T3′ is 2′-F, q 6  is 1, B4′ is 2′-F, and q 7  is 1; with two phosphorothioate internucleotide linkage modifications within position 1-5 of the sense strand (counting from the 5′-end of the sense strand), 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 of the antisense strand). The RNAi agent also comprises a 5′-PS and a targeting ligand. In one embodiment, the 5′-PS is at the 5′-end of the antisense strand, and the targeting ligand is at the 3′-end of the sense strand. 
     In one embodiment, B1 is 2′-OMe or 2′-F, n 1  is 8, T1 is 2′F, n 2  is 3, B2 is 2′-OMe, n 3  is 7, n 4  is 0, B3 is 2′-OMe, n 5  is 3, B1′ is 2′-OMe or 2′-F, q 1  is 9, T1′ is 2′-F, q 2  is 1, B2′ is 2′-OMe or 2′-F, q 3  is 4, T2′ is 2′-F, q 4  is 2, B3′ is 2′-OMe or 2′-F, q 5  is 5, T3′ is 2′-F, q 6  is 1, B4′ is 2′-F, and q 7  is 1; with two phosphorothioate internucleotide linkage modifications within position 1-5 of the sense strand (counting from the 5′-end of the sense strand), 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 of the antisense strand). The RNAi agent also comprises a 5′-VP (e.g., a 5′-E-VP, 5′-Z-VP, or combination thereof) and a targeting ligand. In one embodiment, the 5′-VP is at the 5′-end of the antisense strand, and the targeting ligand is at the 3′-end of the sense strand. 
     In one embodiment, B1 is 2′-OMe or 2′-F, n 1  is 8, T1 is 2′F, n 2  is 3, B2 is 2′-OMe, n 3  is 7, n 4  is 0, B3 is 2′-OMe, n 5  is 3, B1′ is 2′-OMe or 2′-F, q 1  is 9, T1′ is 2′-F, q 2  is 1, B2′ is 2′-OMe or 2′-F, q 3  is 4, T2′ is 2′-F, q 4  is 2, B3′ is 2′-OMe or 2′-F, q 5  is 5, T3′ is 2′-F, q 6  is 1, B4′ is 2′-F, and q 7  is 1; with two phosphorothioate internucleotide linkage modifications within position 1-5 of the sense strand (counting from the 5′-end of the sense strand), 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 of the antisense strand). The RNAi agent also comprises a 5′-PS 2  and a targeting ligand. In one embodiment, the 5′-PS 2  is at the 5′-end of the antisense strand, and the targeting ligand is at the 3′-end of the sense strand. 
     In one embodiment, B1 is 2′-OMe or 2′-F, n 1  is 8, T1 is 2′F, n 2  is 3, B2 is 2′-OMe, n 3  is 7, n 4  is 0, B3 is 2′-OMe, n 5  is 3, B1′ is 2′-OMe or 2′-F, q 1  is 9, T1′ is 2′-F, q 2  is 1, B2′ is 2′-OMe or 2′-F, q 3  is 4, T2′ is 2′-F, q 4  is 2, B3′ is 2′-OMe or 2′-F, q 5  is 5, T3′ is 2′-F, q 6  is 1, B4′ is 2′-F, and q 7  is 1; with two phosphorothioate internucleotide linkage modifications within position 1-5 of the sense strand (counting from the 5′-end of the sense strand), 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 of the antisense strand). The RNAi agent also comprises a 5′-deoxy-5′-C-malonyl and a targeting ligand. In one embodiment, the 5′-deoxy-5′-C-malonyl is at the 5′-end of the antisense strand, and the targeting ligand is at the 3′-end of the sense strand. 
     In one embodiment, B1 is 2′-OMe or 2′-F, n 1  is 8, T1 is 2′F, n 2  is 3, B2 is 2′-OMe, n 3  is 7, n 4  is 0, B3 is 2′-OMe, n 5  is 3, B1′ is 2′-OMe or 2′-F, q 1  is 9, T1′ is 2′-F, q 2  is 1, B2′ is 2′-OMe or 2′-F, q 3  is 4, q 4  is 0, B3′ is 2′-OMe or 2′-F, q 5  is 7, T3′ is 2′-F, q 6  is 1, B4′ is 2′-F, and q 7  is 1; with two phosphorothioate internucleotide linkage modifications within position 1-5 of the sense strand (counting from the 5′-end of the sense strand), 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 of the antisense strand). The RNAi agent also comprises a 5′-P and a targeting ligand. In one embodiment, the 5′-P is at the 5′-end of the antisense strand, and the targeting ligand is at the 3′-end of the sense strand. 
     In one embodiment, B1 is 2′-OMe or 2′-F, n 1  is 8, T1 is 2′F, n 2  is 3, B2 is 2′-OMe, n 3  is 7, n 4  is 0, B3 is 2′-OMe, n 5  is 3, B1′ is 2′-OMe or 2′-F, q 1  is 9, T1′ is 2′-F, q 2  is 1, B2′ is 2′-OMe or 2′-F, q 3  is 4, q 4  is 0, B3′ is 2′-OMe or 2′-F, q 5  is 7, T3′ is 2′-F, q 6  is 1, B4′ is 2′-F, and q 7  is 1; with two phosphorothioate internucleotide linkage modifications within position 1-5 of the sense strand (counting from the 5′-end of the sense strand), 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 of the antisense strand). The RNAi agent also comprises a 5′-PS and a targeting ligand. In one embodiment, the 5′-PS is at the 5′-end of the antisense strand, and the targeting ligand is at the 3′-end of the sense strand. 
     In one embodiment, B1 is 2′-OMe or 2′-F, n 1  is 8, T1 is 2′F, n 2  is 3, B2 is 2′-OMe, n 3  is 7, n 4  is 0, B3 is 2′-OMe, n 5  is 3, B1′ is 2′-OMe or 2′-F, q 1  is 9, T1′ is 2′-F, q 2  is 1, B2′ is 2′-OMe or 2′-F, q 3  is 4, q 4  is 0, B3′ is 2′-OMe or 2′-F, q 5  is 7, T3′ is 2′-F, q 6  is 1, B4′ is 2′-F, and q 7  is 1; with two phosphorothioate internucleotide linkage modifications within position 1-5 of the sense strand (counting from the 5′-end of the sense strand), 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 of the antisense strand). The RNAi agent also comprises a 5′-VP (e.g., a 5′-E-VP, 5′-Z-VP, or combination thereof) and a targeting ligand. In one embodiment, the 5′-VP is at the 5′-end of the antisense strand, and the targeting ligand is at the 3′-end of the sense strand. 
     In one embodiment, B1 is 2′-OMe or 2′-F, n 1  is 8, T1 is 2′F, n 2  is 3, B2 is 2′-OMe, n 3  is 7, n 4  is 0, B3 is 2′-OMe, n 5  is 3, B1′ is 2′-OMe or 2′-F, q 1  is 9, T1′ is 2′-F, q 2  is 1, B2′ is 2′-OMe or 2′-F, q 3  is 4, q 4  is 0, B3′ is 2′-OMe or 2′-F, q 5  is 7, T3′ is 2′-F, q 6  is 1, B4′ is 2′-F, and q 7  is 1; with two phosphorothioate internucleotide linkage modifications within position 1-5 of the sense strand (counting from the 5′-end of the sense strand), 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 of the antisense strand). The RNAi agent also comprises a 5′-PS 2  and a targeting ligand. In one embodiment, the 5′-PS 2  is at the 5′-end of the antisense strand, and the targeting ligand is at the 3′-end of the sense strand. 
     In one embodiment, B1 is 2′-OMe or 2′-F, n 1  is 8, T1 is 2′F, n 2  is 3, B2 is 2′-OMe, n 3  is 7, n 4  is 0, B3 is 2′-OMe, n 5  is 3, B1′ is 2′-OMe or 2′-F, q 1  is 9, T1′ is 2′-F, q 2  is 1, B2′ is 2′-OMe or 2′-F, q 3  is 4, q 4  is 0, B3′ is 2′-OMe or 2′-F, q 5  is 7, T3′ is 2′-F, q 6  is 1, B4′ is 2′-F, and q 7  is 1; with two phosphorothioate internucleotide linkage modifications within position 1-5 of the sense strand (counting from the 5′-end of the sense strand), 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 of the antisense strand). The RNAi agent also comprises a 5′-deoxy-5′-C-malonyl and a targeting ligand. In one embodiment, the 5′-deoxy-5′-C-malonyl is at the 5′-end of the antisense strand, and the targeting ligand is at the 3′-end of the sense strand. 
     In a particular embodiment, an RNAi agent of the present invention comprises:
         (a) a sense strand having:
           (i) a length of 21 nucleotides;   (ii) an ASGPR ligand attached to the 3′-end, wherein said ASGPR ligand comprises three GalNAc derivatives attached through a trivalent branched linker; and   (iii) 2′-F modifications at positions 1, 3, 5, 7, 9 to 11, 13, 17, 19, and 21, and 2′-OMe modifications at positions 2, 4, 6, 8, 12, 14 to 16, 18, and 20 (counting from the 5′ end);   
           and   (b) an antisense strand having:
           (i) a length of 23 nucleotides;   (ii) 2′-OMe modifications at positions 1, 3, 5, 9, 11 to 13, 15, 17, 19, 21, and 23, and 2′F modifications at positions 2, 4, 6 to 8, 10, 14, 16, 18, 20, and 22 (counting from the 5′ end); and   (iii) phosphorothioate internucleotide linkages between nucleotide positions 21 and 22, and between nucleotide positions 22 and 23 (counting from the 5′ end);   
           wherein the dsRNA agents have a two nucleotide overhang at the 3′-end of the antisense strand, and a blunt end at the 5′-end of the antisense strand.       

     In another particular embodiment, an RNAi agent of the present invention comprises:
         (a) a sense strand having:
           (i) a length of 21 nucleotides;   (ii) an ASGPR ligand attached to the 3′-end, wherein said ASGPR ligand comprises three GalNAc derivatives attached through a trivalent branched linker;   (iii) 2′-F modifications at positions 1, 3, 5, 7, 9 to 11, 13, 15, 17, 19, and 21, and 2′-OMe modifications at positions 2, 4, 6, 8, 12, 14, 16, 18, and 20 (counting from the 5′ end); and   (iv) phosphorothioate internucleotide linkages between nucleotide positions 1 and 2, and between nucleotide positions 2 and 3 (counting from the 5′ end);   
           and   (b) an antisense strand having:
           (i) a length of 23 nucleotides;   (ii) 2′-OMe modifications at positions 1, 3, 5, 7, 9, 11 to 13, 15, 17, 19, and 21 to 23, and 2′F modifications at positions 2, 4, 6, 8, 10, 14, 16, 18, and 20 (counting from the 5′ end); and   (iii) 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 (counting from the 5′ end);
 
wherein the RNAi agents have a two nucleotide overhang at the 3′-end of the antisense strand, and a blunt end at the 5′-end of the antisense strand.
   
               

     In another particular embodiment, a RNAi agent of the present invention comprises:
         (a) a sense strand having:
           (i) a length of 21 nucleotides;   (ii) an ASGPR ligand attached to the 3′-end, wherein said ASGPR ligand comprises three GalNAc derivatives attached through a trivalent branched linker;   (iii) 2′-OMe modifications at positions 1 to 6, 8, 10, and 12 to 21, 2′-F modifications at positions 7, and 9, and a deoxy-nucleotide (e.g. dT) at position 11 (counting from the 5′ end); and   (iv) phosphorothioate internucleotide linkages between nucleotide positions 1 and 2, and between nucleotide positions 2 and 3 (counting from the 5′ end);   
           and   (b) an antisense strand having:
           (i) a length of 23 nucleotides;   (ii) 2′-OMe modifications at positions 1, 3, 7, 9, 11, 13, 15, 17, and 19 to 23, and 2′-F modifications at positions 2, 4 to 6, 8, 10, 12, 14, 16, and 18 (counting from the 5′ end); and   (iii) 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 (counting from the 5′ end);
 
wherein the RNAi agents have a two nucleotide overhang at the 3′-end of the antisense strand, and a blunt end at the 5′-end of the antisense strand.
   
               

     In another particular embodiment, a RNAi agent of the present invention comprises:
         (a) a sense strand having:
           (i) a length of 21 nucleotides;   (ii) an ASGPR ligand attached to the 3′-end, wherein said ASGPR ligand comprises three GalNAc derivatives attached through a trivalent branched linker;   (iii) 2′-OMe modifications at positions 1 to 6, 8, 10, 12, 14, and 16 to 21, and 2′-F modifications at positions 7, 9, 11, 13, and 15; and   (iv) phosphorothioate internucleotide linkages between nucleotide positions 1 and 2, and between nucleotide positions 2 and 3 (counting from the 5′ end);   
           and   (b) an antisense strand having:
           (i) a length of 23 nucleotides;   (ii) 2′-OMe modifications at positions 1, 5, 7, 9, 11, 13, 15, 17, 19, and 21 to 23, and 2′-F modifications at positions 2 to 4, 6, 8, 10, 12, 14, 16, 18, and 20 (counting from the 5′ end); and   (iii) 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 (counting from the 5′ end);
 
wherein the RNAi agents have a two nucleotide overhang at the 3′-end of the antisense strand, and a blunt end at the 5′-end of the antisense strand.
   
               

     In another particular embodiment, a RNAi agent of the present invention comprises:
         (a) a sense strand having:
           (i) a length of 21 nucleotides;   (ii) an ASGPR ligand attached to the 3′-end, wherein said ASGPR ligand comprises three GalNAc derivatives attached through a trivalent branched linker;   (iii) 2′-OMe modifications at positions 1 to 9, and 12 to 21, and 2′-F modifications at positions 10, and 11; and   (iv) phosphorothioate internucleotide linkages between nucleotide positions 1 and 2, and between nucleotide positions 2 and 3 (counting from the 5′ end);   
           and   (b) an antisense strand having:
           (i) a length of 23 nucleotides;   (ii) 2′-OMe modifications at positions 1, 3, 5, 7, 9, 11 to 13, 15, 17, 19, and 21 to 23, and 2′-F modifications at positions 2, 4, 6, 8, 10, 14, 16, 18, and 20 (counting from the 5′ end); and   (iii) 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 (counting from the 5′ end);
 
wherein the RNAi agents have a two nucleotide overhang at the 3′-end of the antisense strand, and a blunt end at the 5′-end of the antisense strand.
   
               

     In another particular embodiment, a RNAi agent of the present invention comprises:
         (a) a sense strand having:
           (i) a length of 21 nucleotides;   (ii) an ASGPR ligand attached to the 3′-end, wherein said ASGPR ligand comprises three GalNAc derivatives attached through a trivalent branched linker;   (iii) 2′-F modifications at positions 1, 3, 5, 7, 9 to 11, and 13, and 2′-OMe modifications at positions 2, 4, 6, 8, 12, and 14 to 21; and   (iv) phosphorothioate internucleotide linkages between nucleotide positions 1 and 2, and between nucleotide positions 2 and 3 (counting from the 5′ end);   
           and   (b) an antisense strand having:
           (i) a length of 23 nucleotides;   (ii) 2′-OMe modifications at positions 1, 3, 5 to 7, 9, 11 to 13, 15, 17 to 19, and 21 to 23, and 2′-F modifications at positions 2, 4, 8, 10, 14, 16, and 20 (counting from the 5′ end); and   (iii) 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 (counting from the 5′ end);
 
wherein the RNAi agents have a two nucleotide overhang at the 3′-end of the antisense strand, and a blunt end at the 5′-end of the antisense strand.
   
               

     In another particular embodiment, a RNAi agent of the present invention comprises:
         (a) a sense strand having:
           (i) a length of 21 nucleotides;   (ii) an ASGPR ligand attached to the 3′-end, wherein said ASGPR ligand comprises three GalNAc derivatives attached through a trivalent branched linker;   (iii) 2′-OMe modifications at positions 1, 2, 4, 6, 8, 12, 14, 15, 17, and 19 to 21, and 2′-F modifications at positions 3, 5, 7, 9 to 11, 13, 16, and 18; and   (iv) phosphorothioate internucleotide linkages between nucleotide positions 1 and 2, and between nucleotide positions 2 and 3 (counting from the 5′ end);   
           and   (b) an antisense strand having:
           (i) a length of 25 nucleotides;   (ii) 2′-OMe modifications at positions 1, 4, 6, 7, 9, 11 to 13, 15, 17, and 19 to 23, 2′-F modifications at positions 2, 3, 5, 8, 10, 14, 16, and 18, and desoxy-nucleotides (e.g. dT) at positions 24 and 25 (counting from the 5′ end); and   (iii) 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 (counting from the 5′ end);
 
wherein the RNAi agents have a four nucleotide overhang at the 3′-end of the antisense strand, and a blunt end at the 5′-end of the antisense strand.
   
               

     In another particular embodiment, a RNAi agent of the present invention comprises:
         (a) a sense strand having:
           (i) a length of 21 nucleotides;   (ii) an ASGPR ligand attached to the 3′-end, wherein said ASGPR ligand comprises three GalNAc derivatives attached through a trivalent branched linker;   (iii) 2′-OMe modifications at positions 1 to 6, 8, and 12 to 21, and 2′-F modifications at positions 7, and 9 to 11; and   (iv) phosphorothioate internucleotide linkages between nucleotide positions 1 and 2, and between nucleotide positions 2 and 3 (counting from the 5′ end);   
           and   (b) an antisense strand having:
           (i) a length of 23 nucleotides;   (ii) 2′-OMe modifications at positions 1, 3 to 5, 7, 8, 10 to 13, 15, and 17 to 23, and 2′-F modifications at positions 2, 6, 9, 14, and 16 (counting from the 5′ end); and   (iii) 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 (counting from the 5′ end);
 
wherein the RNAi agents have a two nucleotide overhang at the 3′-end of the antisense strand, and a blunt end at the 5′-end of the antisense strand.
   
               

     In another particular embodiment, a RNAi agent of the present invention comprises:
         (a) a sense strand having:
           (i) a length of 21 nucleotides;   (ii) an ASGPR ligand attached to the 3′-end, wherein said ASGPR ligand comprises three GalNAc derivatives attached through a trivalent branched linker;   (iii) 2′-OMe modifications at positions 1 to 6, 8, and 12 to 21, and 2′-F modifications at positions 7, and 9 to 11; and   (iv) phosphorothioate internucleotide linkages between nucleotide positions 1 and 2, and between nucleotide positions 2 and 3 (counting from the 5′ end);   
           and   (b) an antisense strand having:
           (i) a length of 23 nucleotides;   (ii) 2′-OMe modifications at positions 1, 3 to 5, 7, 10 to 13, 15, and 17 to 23, and 2′-F modifications at positions 2, 6, 8, 9, 14, and 16 (counting from the 5′ end); and   (iii) 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 (counting from the 5′ end);
 
wherein the RNAi agents have a two nucleotide overhang at the 3′-end of the antisense strand, and a blunt end at the 5′-end of the antisense strand.
   
               

     In another particular embodiment, a RNAi agent of the present invention comprises:
         (a) a sense strand having:
           (i) a length of 19 nucleotides;   (ii) an ASGPR ligand attached to the 3′-end, wherein said ASGPR ligand comprises three GalNAc derivatives attached through a trivalent branched linker;   (iii) 2′-OMe modifications at positions 1 to 4, 6, and 10 to 19, and 2′-F modifications at positions 5, and 7 to 9; and   (iv) phosphorothioate internucleotide linkages between nucleotide positions 1 and 2, and between nucleotide positions 2 and 3 (counting from the 5′ end);   
           and   (b) an antisense strand having:
           (i) a length of 21 nucleotides;   (ii) 2′-OMe modifications at positions 1, 3 to 5, 7, 10 to 13, 15, and 17 to 21, and 2′-F modifications at positions 2, 6, 8, 9, 14, and 16 (counting from the 5′ end); and   (iii) phosphorothioate internucleotide linkages between nucleotide positions 1 and 2, between nucleotide positions 2 and 3, between nucleotide positions 19 and 20, and between nucleotide positions 20 and 21 (counting from the 5′ end);
 
wherein the RNAi agents have a two nucleotide overhang at the 3′-end of the antisense strand, and a blunt end at the 5′-end of the antisense strand.
   
               

     In certain embodiments, the iRNA for use in the methods of the invention is an agent selected from agents listed in Table 2, Table 3, Table 6 or Table 8. These agents may further comprise a ligand. 
     III. 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). In other embodiments, the ligand is 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 preferred 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. Preferred ligands do 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, N-acetylglucosamine, N-acetylgalactosamine, or hyaluronic acid); or a lipid. The ligand can 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 alpha 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, vitamin A, 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, 03-(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 hepatic cell. Ligands can 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, taxol, 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. 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 methods 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 iRNAs 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 preferably binds a serum protein, e.g., 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 inhibit, e.g., control 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. Preferably, it binds HSA with a sufficient affinity such that the conjugate will be preferably distributed to a non-kidney tissue. However, it is preferred that the affinity not be so strong that the HSA-ligand binding cannot be reversed. 
     In other embodiments, the lipid based ligand binds HSA weakly or not at all, such that the conjugate will be preferably distributed to the kidney. 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 target cells such as liver cells. Also included are HSA and low density lipoprotein (LDL). 
     B. Cell Permeation Agents 
     In another aspect, the ligand is a cell-permeation agent, preferably a helical cell-permeation agent. Preferably, 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 preferably an alpha-helical agent, which preferably has 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: 25). An RFGF analogue (e.g., amino acid sequence AALLPVLLAAP (SEQ ID NO:26) 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:27) and the  Drosophila  Antennapedia protein (RQIKIWFQNRRMKWKK (SEQ ID NO:28) 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). Examples of a peptide or peptidomimetic tethered to a dsRNA agent via an incorporated monomer unit for cell targeting purposes is 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. 
     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 is 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 trisaccharides include sugars having two or three monosaccharide units (e.g., C5, C6, C7, or C8). 
     In certain embodiments, a carbohydrate conjugate for use in the compositions and methods of the invention is a monosaccharide. 
     In one embodiment, a carbohydrate conjugate for use in the compositions and methods of the invention is selected from the group consisting of: 
     
       
         
         
             
             
         
       
       
         
         
             
             
         
       
       
         
         
             
             
         
       
       
         
         
             
             
         
       
       
         
         
             
             
         
       
       
         
         
             
             
         
       
       
         
         
             
             
         
       
       
         
         
             
             
         
       
       
         
         
             
             
         
       
     
     In another embodiment, a carbohydrate conjugate for use in the compositions and methods of the invention is a monosaccharide. In one embodiment, 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 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 one embodiment, 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 another embodiment, 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 PCT Publication Nos. 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 NRB, 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, or substituted or unsubstituted heterocyclic; where R8 is hydrogen, acyl, aliphatic, or substituted aliphatic. In one embodiment, the linker is about 1-24 atoms, 2-24, 3-24, 4-24, 5-24, 6-24, 6-18, 7-18, 8-18, 7-17, 8-17, 6-16, 7-17, 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 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 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 other 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)-O—, —S—P(O)(Rk)-S—, —O—P(S)(Rk)-S—. Preferred embodiments are —O—P(O)(OH)—O—, —O—P(S)(OH)—O—, —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—, —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—, and —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 other 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, 5.5, 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 Linking Groups 
     In other 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 Cleaving Groups 
     In yet other embodiments, 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 one embodiment, 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  and 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 dsRNAi 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. 
     IV. Delivery of an iRNA of the Invention 
     The delivery of an iRNA of the invention 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 susceptible to or diagnosed with a SERPINF2 associated disorder, e.g., a disorder associated with thrombosis, e.g., plasminogen deficiency, venous thrombosis, venous thromboembolism, deep vein thrombosis, pulmonary embolism, prosthetic valve thrombosis, post-thrombotic syndrome, atherothrombosis, heart attack, stroke, fibrinolytic disorders, hypofibrinolysis, disfibrinolysis, ischemic stroke, atrial fibrillation, myocardial infarction, peripheral arterial disease, dynamic thrombosis, coronary artery disease, microvascular thrombosis, ischemic brain injury, brain swelling, brain hemorrhage and death after thromboembolic stroke, can be achieved in a number of different ways. For example, delivery may be performed by contacting a cell with an iRNA of the invention either in vitro or in vivo. In vivo delivery may also be performed directly by administering a composition comprising an iRNA, 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 iRNA. 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 an iRNA of the invention (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 iRNA molecule include, for example, biological stability of the delivered molecule, prevention of non-specific effects, and accumulation of the delivered molecule in the target tissue. 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). Modification of the RNA or the pharmaceutical carrier can also permit targeting of the iRNA to the target tissue and avoid undesirable off-target effects. iRNA molecules can be modified by chemical conjugation to lipophilic groups such as cholesterol to enhance cellular uptake and prevent degradation. For example, an iRNA 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). 
     In an alternative embodiment, the iRNA 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 an iRNA molecule (negatively charged) and also enhance interactions at the negatively charged cell membrane to permit efficient uptake of an iRNA by the cell. Cationic lipids, dendrimers, or polymers can either be bound to an iRNA, 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 iRNA. The formation of vesicles or micelles further prevents degradation of the iRNA when administered systemically. Methods for making and administering cationic-iRNA 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 iRNAs include DOTAP (Sorensen, D R., et al (2003), supra; Verma, U N, et al (2003), supra), “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 . August 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, an iRNA forms a complex with cyclodextrin for systemic administration. Methods for administration and pharmaceutical compositions of iRNAs and cyclodextrins can be found in U.S. Pat. No. 7,427,605, which is herein incorporated by reference in its entirety. 
     A. Vector Encoded iRNAs of the Invention 
     iRNA targeting the SERPINF2 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; Skillern, A, et al., International PCT Publication No. WO 00/22113, Conrad, International PCT Publication No. WO 00/22114, and Conrad, U.S. Pat. No. 6,054,299). Expression can be transient (on the order of hours to weeks) or sustained (weeks to 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.,  Proc. Natl. Acad. Sci. USA  (1995) 92:1292). 
     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 an iRNA will generally require regulatory elements, e.g., promoters, enhancers, etc., to ensure the expression of the iRNA in target cells. Other aspects to consider for vectors and constructs are known in the art. 
     V. Pharmaceutical Compositions of the Invention 
     The present invention also includes pharmaceutical compositions and formulations which include the iRNAs of the invention. In one embodiment, provided herein are pharmaceutical compositions containing an iRNA, as described herein, and a pharmaceutically acceptable carrier. The pharmaceutical compositions containing the iRNA are useful for preventing or treating a SERPINF2 associated disorder, e.g., a disorder associated with thrombosis, e.g., plasminogen deficiency, venous thrombosis, venous thromboembolism, deep vein thrombosis, pulmonary embolism, prosthetic valve thrombosis, post-thrombotic syndrome, atherothrombosis, heart attack, stroke, fibrinolytic disorders, hypofibrinolysis, disfibrinolysis, ischemic stroke, atrial fibrillation, myocardial infarction, peripheral arterial disease, dynamic thrombosis, coronary artery disease, microvascular thrombosis, ischemic brain injury, brain swelling, brain hemorrhage and death after thromboembolic stroke. Such pharmaceutical compositions are formulated based on the mode of delivery. One example is compositions that are formulated for systemic administration via parenteral delivery, e.g., by subcutaneous (SC), intramuscular (IM), or intravenous (IV) delivery. The pharmaceutical compositions of the invention may be administered in dosages sufficient to inhibit expression of a SERPINF2 gene. 
     The pharmaceutical compositions of the invention may be administered in dosages sufficient to inhibit expression of a SERPINF2 gene. In general, a suitable dose of an iRNA of the invention will be in the range of about 0.001 to about 200.0 milligrams per kilogram body weight of the recipient per day, generally in the range of about 1 to 50 mg per kilogram body weight per day. Typically, a suitable dose of an iRNA of the invention will be in the range of about 0.1 mg/kg to about 5.0 mg/kg, preferably about 0.3 mg/kg and about 3.0 mg/kg. A repeat-dose regimen may include administration of a therapeutic amount of iRNA on a regular basis, such as every month, once every 3-6 months, or once a year. In certain embodiments, the iRNA is administered about once per month to about once per six months. 
     After an initial treatment regimen, the treatments can be administered on a less frequent basis. Duration of treatment can be determined based on the severity of disease. 
     In other embodiments, a single dose of the pharmaceutical compositions can be long lasting, such that doses are administered at not more than 1, 2, 3, or 4 month intervals. In some embodiments of the invention, a single dose of the pharmaceutical compositions of the invention is administered about once per month. In other embodiments of the invention, a single dose of the pharmaceutical compositions of the invention is administered quarterly (i.e., about every three months). In other embodiments of the invention, a single dose of the pharmaceutical compositions of the invention is administered twice per year (i.e., about once every six months). 
     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 mutations present in the subject, previous treatments, the general health or age of the subject, and other diseases present. Moreover, treatment of a subject with a prophylactically or therapeutically effective amount, as appropriate, of a composition can include a single treatment or a series of treatments. 
     The iRNA can be delivered in a manner to target a particular tissue (e.g., hepatocytes or eyes). 
     Pharmaceutical compositions of the present invention 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. Formulations include those that target the liver. 
     The pharmaceutical formulations of the present invention, 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. 
     A. Additional Formulations 
     i. Emulsions 
     The compositions of the present invention 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 either in the 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. 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). 
     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). 
     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). 
     ii. Microemulsions 
     In one embodiment of the present invention, the compositions of iRNAs 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). 
     iii. Microparticles 
     An iRNA of the invention 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 invention employs various penetration enhancers to effect the efficient delivery of nucleic acids, particularly iRNAs, 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 and their use in manufacture of pharmaceutical compositions and delivery of pharmaceutical agents are well known in the art. 
     v. 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. Such agent are well known in the art. 
     vi. Other Components 
     The compositions of the present invention 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 invention, 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 invention. 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 invention include (a) one or more iRNA and (b) one or more agents which function by a non-iRNA mechanism and which are useful in treating a SERPINF2 associated disorder, e.g., a disorder associated with thrombosis, e.g., plasminogen deficiency, venous thrombosis, venous thromboembolism, deep vein thrombosis, pulmonary embolism, prosthetic valve thrombosis, post-thrombotic syndrome, atherothrombosis, heart attack, stroke, fibrinolytic disorders, hypofibrinolysis, disfibrinolysis, ischemic stroke, atrial fibrillation, myocardial infarction, peripheral arterial disease, dynamic thrombosis, coronary artery disease, microvascular thrombosis, ischemic brain injury, brain swelling, brain hemorrhage and death after thromboembolic stroke. 
     Toxicity and prophylactic efficacy of such compounds can be determined by standard pharmaceutical procedures in cell cultures or experimental animals, e.g., for determining the LD50 (the dose lethal to 50% of the population) and the ED50 (the dose prophylactically 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 LD50/ED50. 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 invention lies generally within a range of circulating concentrations that include the ED50, preferably an ED80 or ED90, 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 invention, the prophylactically 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 IC50 (i.e., the concentration of the test compound which achieves a half-maximal inhibition of symptoms) or higher levels of inhibition 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 iRNAs featured in the invention can be administered in combination with other known agents used for the prevention or treatment of a SERPINF2 associated disorder, e.g., a disorder associated with thrombosis, e.g., plasminogen deficiency, venous thrombosis, venous thromboembolism, deep vein thrombosis, pulmonary embolism, prosthetic valve thrombosis, post-thrombotic syndrome, atherothrombosis, heart attack, stroke, fibrinolytic disorders, hypofibrinolysis, disfibrinolysis, ischemic stroke, atrial fibrillation, myocardial infarction, peripheral arterial disease, dynamic thrombosis, coronary artery disease, microvascular thrombosis, ischemic brain injury, brain swelling, brain hemorrhage and death after thromboembolic stroke. In any event, the administering physician can adjust the amount and timing of iRNA administration on the basis of results observed using standard measures of efficacy known in the art or described herein. 
     VI. Methods for Inhibiting SERPINF2 Expression 
     The present invention also provides methods of inhibiting expression of a SERPINF2 gene in a cell. The methods include contacting a cell with an RNAi agent, e.g., double stranded RNA agent, in an amount effective to inhibit expression of SERPINF2 in the cell, thereby inhibiting expression of SERPINF2 in the cell. 
     Contacting of a cell with an iRNA, e.g., a double stranded RNA agent, may be done in vitro or in vivo. Contacting a cell in vivo with the iRNA includes contacting a cell or group of cells within a subject, e.g., a human subject, with the iRNA. 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 preferred embodiments, the targeting ligand is a carbohydrate moiety, e.g., a GalNAc 3  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. 
     The phrase “inhibiting expression of a SERPINF2” is intended to refer to inhibition of expression of any SERPINF2 gene (such as, e.g., a mouse SERPINF2 gene, a rat SERPINF2 gene, a monkey SERPINF2 gene, or a huma SERPINF2 gene) as well as variants or mutants of a SERPINF2 gene. Thus, the SERPINF2 gene may be a wild-type SERPINF2 gene, a mutant SERPINF2 gene, or a transgenic SERPINF2 gene in the context of a genetically manipulated cell, group of cells, or organism. 
     “Inhibiting expression of a SERPINF2 gene” includes any level of inhibition of a SERPINF2 gene, e.g., at least partial suppression of the expression of a SERPINF2 gene. The expression of the SERPINF2 gene may be assessed based on the level, or the change in the level, of any variable associated with SERPINF2 gene expression, e.g., SERPINF2 mRNA level or SERPINF2 protein level. This level may be assessed in an individual cell or in a group of cells, including, for example, a sample derived from a subject. It is understood that SERPINF2 is expressed predominantly in the liver, but also in the brain, gall bladder, heart, and kidney, and is present in circulation. 
     Inhibition may be assessed by a decrease in an absolute or relative level of one or more variables that are associated with SERPINF2 expression 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 invention, expression of a SERPINF2 gene is inhibited by at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, or 95%, or to below the level of detection of the assay. In preferred embodiments, expression of a SERPINF2 gene is inhibited by at least 70%. It is further understood that inhibition of SERPINF2 expression in certain tissues, e.g., in liver, without a significant inhibition of expression in other tissues, e.g., brain, may be desirable. In preferred embodiments, expression level is determined using the assay method provided in Example 2 with a 10 nM siRNA concentration in the appropriate species matched cell line. 
     In certain embodiments, inhibition of expression in vivo is determined by knockdown of the human gene in a rodent expressing the human gene, e.g., an AAV-infected mouse expressing the human target gene (i.e., SERPINF2), e.g., when administered a single dose at 3 mg/kg at the nadir of RNA expression. Knockdown of expression of an endogenous gene in a model animal system can also be determined, e.g., after administration of a single dose at 3 mg/kg at the nadir of RNA expression. Such systems are useful when the nucleic acid sequence of the human gene and the model animal gene are sufficiently close such that the human iRNA provides effective knockdown of the model animal gene. RNA expression in liver is determined using the PCR methods provided in Example 2 and Example 3. 
     Inhibition of the expression of a SERPINF2 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 a SERPINF2 gene is transcribed and which has or have been treated (e.g., by contacting the cell or cells with an iRNA of the invention, or by administering an iRNA of the invention to a subject in which the cells are or were present) such that the expression of a SERPINF2 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 an iRNA or not treated with an iRNA targeted to the gene of interest). In preferred embodiments, the inhibition is assessed by the method provided in Example 2 using a 10 nM siRNA concentration in the species matched cell line and expressing the level of mRNA in treated cells as a percentage of the level of mRNA in control cells, using the following formula: 
     
       
         
           
             
               
                 
                   
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                       cells 
                     
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     In other embodiments, inhibition of the expression of a SERPINF2 gene may be assessed in terms of a reduction of a parameter that is functionally linked to SERPINF2 gene expression, e.g., SERPINF2 protein level in blood or serum from a subject. SERPINF2 gene silencing may be determined in any cell expressing SERPINF2, either endogenous or heterologous from an expression construct, and by any assay known in the art. 
     Inhibition of the expression of a SERPINF2 protein may be manifested by a reduction in the level of the SERPINF2 protein that is expressed by a cell or group of cells or in a subject sample (e.g., the level of protein in a blood sample derived from a subject). As explained above, for the assessment of mRNA 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, or the change in the level of protein in a subject sample, e.g., blood or serum derived therefrom. 
     A control cell, a group of cells, or subject sample that may be used to assess the inhibition of the expression of a SERPINF2 gene includes a cell, group of cells, or subject sample that has not yet been contacted with an RNAi agent of the invention. For example, the control cell, group of cells, or subject sample may be derived from an individual subject (e.g., a human or animal subject) prior to treatment of the subject with an RNAi agent or an appropriately matched population control. 
     The level of SERPINF2 mRNA that is expressed by a cell or group of cells may be determined using any method known in the art for assessing mRNA expression. In one embodiment, the level of expression of SERPINF2 in a sample is determined by detecting a transcribed polynucleotide, or portion thereof, e.g., mRNA of the SERPINF2 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. 
     In some embodiments, the level of expression of SERPINF2 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 SERPINF2. 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 mRNA levels involves contacting the isolated mRNA with a nucleic acid molecule (probe) that can hybridize to SERPINF2 mRNA. In one embodiment, the mRNA is immobilized on a solid surface and contacted with a probe, for example by running the isolated mRNA on an agarose gel and transferring the mRNA from the gel to a membrane, such as nitrocellulose. In an alternative embodiment, the probe(s) are immobilized on a solid surface and the mRNA is contacted with the probe(s), for example, in an Affymetrix® gene chip array. A skilled artisan can readily adapt known mRNA detection methods for use in determining the level of SERPINF2 mRNA. 
     An alternative method for determining the level of expression of SERPINF2 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 invention, the level of expression of SERPINF2 is determined by quantitative fluorogenic RT-PCR (i.e., the TaqMan™ System). In preferred embodiments, expression level is determined by the method provided in Example 2 using a 10 nM siRNA concentration in the species matched cell line. 
     The expression levels of SERPINF2 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 SERPINF2 expression level may also comprise using nucleic acid probes in solution. 
     In preferred embodiments, the level of mRNA expression is assessed using branched DNA (bDNA) assays or real time PCR (qPCR). The use of these methods is described and exemplified in the Examples presented herein. In preferred embodiments, expression level is determined by the method provided in Example 2 using a 10 nM siRNA concentration in the species matched cell line. 
     The level of SERPINF2 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. 
     In some embodiments, the efficacy of the methods of the invention are assessed by a decrease in SERPINF2 mRNA or protein level (e.g., in a liver biopsy). 
     In some embodiments of the methods of the invention, the iRNA is administered to a subject such that the iRNA is delivered to a specific site within the subject. The inhibition of expression of SERPINF2 may be assessed using measurements of the level or change in the level of SERPINF2 mRNA or SERPINF2 protein in a sample derived from fluid or tissue from the specific site within the subject (e.g., liver or blood). 
     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. 
     VII. Prohylactic and Treatment Methods of the Invention 
     The present invention also provides methods of using an iRNA of the invention or a composition containing an iRNA of the invention to inhibit expression of SERPINF2, thereby preventing or treating a a SERPINF2 associated disorder, e.g., a disorder associated with thrombosis, e.g., plasminogen deficiency, venous thrombosis, venous thromboembolism, deep vein thrombosis, pulmonary embolism, prosthetic valve thrombosis, post-thrombotic syndrome, atherothrombosis, heart attack, stroke, fibrinolytic disorders, hypofibrinolysis, disfibrinolysis, ischemic stroke, atrial fibrillation, myocardial infarction, peripheral arterial disease, dynamic thrombosis, coronary artery disease, microvascular thrombosis, ischemic brain injury, brain swelling, brain hemorrhage and death after thromboembolic stroke. 
     In the methods of the invention the cell may be contacted with the siRNA in vitro or in vivo, i.e., the cell may be within a subject. 
     A cell suitable for treatment using the methods of the invention may be any cell that expresses a SERPINF2 gene, e.g., a liver cell, an eye cell, a brain cell, a gall bladder cell, a heart cell, or a kidney cell, but preferably a liver cell. A cell suitable for use in the methods of the invention may be a mammalian cell, e.g., a primate cell (such as a human cell, including human cell in a chimeric non-human animal, or a non-human primate cell, e.g., a monkey cell or a chimpanzee cell), or a non-primate cell. In certain embodiments, the cell is a human cell, e.g., a human liver cell. In the methods of the invention, SERPINF2 expression is inhibited in the cell by at least 50, 55, 60, 65, 70, 75, 80, 85, 90, or 95, or to a level below the level of detection of the assay. 
     The in vivo methods of the invention may include administering to a subject a composition containing an iRNA, where the iRNA includes a nucleotide sequence that is complementary to at least a part of an RNA transcript of the SERPINF2 gene of the mammal to which the RNAi agent is to be administered. 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 intramuscular injection. 
     In one aspect, the present invention also provides methods for inhibiting the expression of a SERPINF2 gene in a mammal. The methods include administering to the mammal a composition comprising a dsRNA that targets a SERPINF2 gene in a cell of the mammal and maintaining the mammal for a time sufficient to obtain degradation of the mRNA transcript of the SERPINF2 gene, thereby inhibiting expression of the SERPINF2 gene in the cell. Reduction in gene expression can be assessed by any methods known in the art and by methods, e.g. qRT-PCR, described herein, e.g., in Example 2. Reduction in protein production can be assessed by any methods known it the art, e.g. ELISA. In certain embodiments, a puncture liver biopsy sample serves as the tissue material for monitoring the reduction in the SERPINF2 gene or protein expression. In other embodiments, a blood sample serves as the subject sample for monitoring the reduction in the SERPINF2 protein expression. The present invention further provides methods of treatment in a subject in need thereof, e.g., a subject diagnosed with a SERPINF2 associated disorder, e.g., a disorder associated with thrombosis, e.g., plasminogen deficiency, venous thrombosis, venous thromboembolism, deep vein thrombosis, pulmonary embolism, prosthetic valve thrombosis, post-thrombotic syndrome, atherothrombosis, heart attack, stroke, fibrinolytic disorders, hypofibrinolysis, disfibrinolysis, ischemic stroke, atrial fibrillation, myocardial infarction, peripheral arterial disease, dynamic thrombosis, coronary artery disease, microvascular thrombosis, ischemic brain injury, brain swelling, brain hemorrhage and death after thromboembolic stroke. In some embodiments, the SERPINF2 associated disorder is a type I plasminogen deficiency. In other embodiments, the SERPINF2 associated disorder is a type II plasminogen deficiency. 
     The present invention further provides methods of prophylaxis in a subject in need thereof. The treatment methods of the invention include administering an iRNA of the invention to a subject, e.g., a subject that would benefit from a reduction of SERPINF2 expression, such as a subject suffering from a SERPINF2 associated disorder, in a prophylactically effective amount of an iRNA targeting a SERPINF2 gene or a pharmaceutical composition comprising an iRNA targeting a SERPINF2 gene. 
     An iRNA of the invention may be administered as a “free iRNA.” A free iRNA is administered in the absence of a pharmaceutical composition. The naked iRNA 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 iRNA can be adjusted such that it is suitable for administering to a subject. 
     Alternatively, an iRNA of the invention may be administered as a pharmaceutical composition, such as a dsRNA liposomal formulation. 
     Subjects that would benefit from an inhibition of SERPINF2 gene expression are subjects suffering or prone to suffering from a SERPINF2 associated disorder, e.g., subjects susceptible to or diagnosed with, e.g., a disorder associated with thrombosis, e.g., plasminogen deficiency, venous thrombosis, venous thromboembolism, deep vein thrombosis, pulmonary embolism, prosthetic valve thrombosis, post-thrombotic syndrome, atherothrombosis, heart attack, stroke, fibrinolytic disorders, hypofibrinolysis, disfibrinolysis, ischemic stroke, atrial fibrillation, myocardial infarction, peripheral arterial disease, dynamic thrombosis, coronary artery disease, microvascular thrombosis, ischemic brain injury, brain swelling, brain hemorrhage and death after thromboembolic stroke. In some embodiments, the SERPINF2 associated disorder is a type I plasminogen deficiency. In other embodiments, the SERPINF2 associated disorder is a type II plasminogen deficiency. 
     In an embodiment, the method includes administering a composition featured herein such that expression of the target SERPINF2 gene is decreased, such as for about 1, 2, 3, 4, 5, 6, 1-6, 1-3, or 3-6 months per dose. In certain embodiments, the composition is administered once every 3-6 months. 
     Preferably, the iRNAs useful for the methods and compositions featured herein specifically target RNAs (primary or processed) of the target SERPINF2 gene. Compositions and methods for inhibiting the expression of these genes using iRNAs can be prepared and performed as described herein. 
     Administration of the iRNA according to the methods of the invention may result prevention or treatment of a SERPINF2 associated disorder, e.g., a disorder associated with thrombosis, e.g., plasminogen deficiency, venous thrombosis, venous thromboembolism, deep vein thrombosis, pulmonary embolism, prosthetic valve thrombosis, post-thrombotic syndrome, atherothrombosis, heart attack, stroke, fibrinolytic disorders, hypofibrinolysis, disfibrinolysis, ischemic stroke, atrial fibrillation, myocardial infarction, peripheral arterial disease, dynamic thrombosis, coronary artery disease, microvascular thrombosis, ischemic brain injury, brain swelling, brain hemorrhage and death after thromboembolic stroke. In some embodiments, the SERPINF2 associated disorder is a type I plasminogen deficiency. In other embodiments, the SERPINF2 associated disorder is a type II plasminogen deficiency. 
     Subjects can be administered a therapeutic amount of iRNA, such as about 0.01 mg/kg to about 200 mg/kg. 
     The iRNA is preferably administered subcutaneously, i.e., by subcutaneous injection. One or more injections may be used to deliver the desired dose of iRNA 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 iRNA on a regular basis, such as once per month to once a year. In certain embodiments, the iRNA is administered about once per month to about once every three months, or about once every three months to about once every six months. 
     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 Informal 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 
     A set of siRNAs targeting the human SERPINF2, “serpin family F member 2” (human: NCBI refseqID NM_000934.3; NCBI GeneID: 5345) as well as SERPINF2 orthologs ( Mus musculus : NM_008878.2 NCBI GeneID: 18816) were designed using custom R and Python scripts. The human NM_000934 REFSEQ mRNA, version 3, has a length of 2284 bases. The mouse NM_008878 REFSEQ mRNA, version 2, has a length of 2199 bases. The rationale and method for the set of siRNA designs is as follows: the predicted efficacy for every potential 19mer siRNA from position 10 through the 3′ end of the transcript was determined with a random forest model derived from the direct measure of mRNA knockdown from several thousand distinct siRNA designs targeting a diverse set of vertebrate genes. For each strand of the siRNA, a custom Python script was used in a brute force search to measure the number and positions of mismatches between the siRNA and all potential alignments in the target species transcriptome. Extra weight was given to mismatches in the seed region, defined here as positions 2-9 of the antisense oligonucleotide, as well the cleavage site of the siRNA, defined here as positions 10-11 of the antisense oligonucleotide. The relative weight of the mismatches was 2.8, 1.2, 1 for seed mismatches, cleavage site, and other positions up through antisense position 19. Mismatches in the first position were ignored. A specificity score was calculated for each strand by summing the value of each weighted mismatch. Preference was given to siRNAs whose antisense score in human was &gt;=2.2 and predicted efficacy was &gt;=50% knockdown. 
     A detailed list of the unmodified SERPINF2 sense and antisense strand nucleotide sequences is shown in Table 2. A detailed list of the modified SERPINF2 sense and antisense strand nucleotide sequences is shown in Table 3. 
     siRNA Synthesis 
     siRNAs were synthesized and annealed using routine methods known in the art. 
     Briefly, siRNA sequences were synthesized at 1 μmol scale on a Mermade 192 synthesizer (BioAutomation) using the solid support mediated phosphoramidite chemistry. The solid support was controlled pore glass (500 A) loaded with custom GalNAc ligand or universal solid support (AM biochemical). Ancillary synthesis reagents, 2′-F and 2′-O-Methyl RNA and deoxy phosphoramidites were obtained from Thermo-Fisher (Milwaukee, Wis.) and Hongene (China). 2′F 2′-O-Methyl, GNA (glycol nucleic acids), 5′phosphate and other modifications were introduced using the corresponding phosphoramidites. Synthesis of 3′ GalNAc conjugated single strands was performed on a GalNAc modified CPG support. Custom CPG universal solid support was used for the synthesis of antisense single strands. Coupling time for all phosphoramidites (100 mM in acetonitrile) was 5 min employing 5-Ethylthio-1H-tetrazole (ETT) as activator (0.6 M in acetonitrile). Phosphorothioate linkages were generated using a 50 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 (1:1 v/v). Oxidation time was 3 minutes. All sequences were synthesized with final removal of the DMT group (“DMT off”). 
     Upon completion of the solid phase synthesis, oligoribonucleotides were cleaved from the solid support and deprotected in sealed 96 deep well plates using 200 μL Aqueous Methylamine reagents at 60° C. for 20 minutes. For sequences containing 2′ ribo residues (2′-OH) that are protected with a tert-butyl dimethyl silyl (TBDMS) group, a second step deprotection was performed using TEA.3HF (triethylamine trihydro fluoride) reagent. To the methylamine deprotection solution, 200 uL of dimethyl sulfoxide (DMSO) and 300 ul TEA.3HF reagent was added and the solution was incubated for additional 20 min at 60° C. At the end of cleavage and deprotection step, the synthesis plate was allowed to come to room temperature and was precipitated by addition of 1 mL of acetontile:ethanol mixture (9:1). The plates were cooled at −80 C for 2 hrs, superanatant decanted carefully with the aid of a multi channel pipette. The oligonucleotide pellet was re-suspended in 20 mM NaOAc buffer and were desalted using a 5 mL HiTrap size exclusion column (GE Healthcare) on an AKTA Purifier System equipped with an A905 autosampler and a Frac 950 fraction collector. Desalted samples were collected in 96-well plates. Samples from each sequence were analyzed by LC-MS to confirm the identity, UV (260 nm) for quantification and a selected set of samples by IEX chromatography to determine purity. 
     Annealing of single strands was performed on a Tecan liquid handling robot. Equimolar mixture of sense and antisense single strands were combined and annealed in 96 well plates. After combining the complementary single strands, the 96-well plate was sealed tightly and heated in an oven at 100° C. for 10 minutes and allowed to come slowly to room temperature over a period 2-3 hours. The concentration of each duplex was normalized to 10 μM in 1×PBS and then submitted for in vitro screening assays. 
     Example 2. In Vitro Screening Methods 
     Cell Culture and Transfections: 
     Hep3B(ATCC) and Primary Mouse Hepactocytes cells (BioIVT) were transfected by adding 5 ul of Opti-MEM plus 0.1 ul of Lipofectamine RNAiMax per well (Invitrogen, Carlsbad Calif. cat #13778-150) to 5.1 ul of siRNA duplexes per well into a 384-well plate and incubated at room temperature for 15 minutes. 40 ul of Eagle&#39;s Minimum Essential Medium (ATCC Cat #30-2003) and InVitroGRO™ CP Rodent Medium (BioIVT Cat #Z990028) respectively, containing ˜5×10 3  cells were then added to the siRNA mixture. Cells were incubated for 24 hours prior to RNA purification. Single dose experiments were performed at 10 nM. 
     Total RNA Isolation Using DYNABEADS mRNA Isolation Kit: 
     RNA was isolated using an automated protocol on a BioTek-EL406 platform using DYNABEADs (Invitrogen, cat #61012). Briefly, 70 ul of Lysis/Binding Buffer and 10 ul of lysis buffer containing 3 ul of magnetic beads were added to the plate with cells. Plates were incubated on an electromagnetic shaker for 10 minutes at room temperature and then magnetic beads were captured and the supernatant was removed. Bead-bound RNA was then washed 2 times with 150 ul Wash Buffer A and once with Wash Buffer B. Beads were then washed with 150 ul Elution Buffer, re-captured and supernatant removed. 
     cDNA Synthesis Using ABI High Capacity cDNA Reverse Transcription Kit (Applied Biosystems, Foster City, Calif., Cat #4368813): 
     Twelve μl of a master mix containing 1.2 ul 10× Buffer, 0.48 ul 25× dNTPs, 1.2 ul 10× Random primers, 0.6 ul Reverse Transcriptase, 0.6 ul RNase inhibitor and 7.92 ul of H 2 O per reaction was added to RNA isolated above. Plates were sealed, mixed, and incubated on an electromagnetic shaker for 10 minutes at room temperature, followed by 2 h 37° C. 
     Real Time PCR: 
     Two μl of cDNA were added to a master mix containing 0.5 μl of Human GAPDH TaqMan Probe (4326317E), and 0.5 μl SERPINF2 human probe (Hs00168686_m1) and 5 μl Lightcycler 480 probe master mix (Roche Cat #04887301001) per well in a 384 well plates (Roche cat #04887301001). For the mouse screen the master mix contained 0.5 μl of mouse GAPDH TaqMan Probe (4352339E), and 0.5 μl SERPINF2 mouse probe (Mm00435868_m1). Real time PCR was done in a LightCycler480 Real Time PCR system (Roche). Each duplex was tested at least two times and data were normalized to cells transfected with a non-targeting control siRNA. To calculate relative fold change, real time data were analyzed using the ΔΔCt method and normalized to assays performed with cells transfected with a non-targeting control siRNA. Results from the screening are shown in Table 4 and Table 5 below. 
     
       
         
           
               
             
               
                 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 
               
               
                 L96 
                 N-[tris(GalNAc-alkyl)-amidodecanoyl)]-4-hydroxyprolinol 
               
               
                   
                 (Hyp-(GalNAc-alkyl)3) 
               
               
                 Y34 
                 2-hydroxymethyl-tetrahydrofurane-4-methoxy-3-phosphate  
               
               
                   
                 (abasic 2′-OMefuranose) 
               
               
                 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 
               
               
                 (Aam) 
                 2{grave over ( )}-O-(N-methylacetamide)adenosine-3{grave over ( )}-phosphate 
               
               
                 (Aams) 
                 2{grave over ( )}-O-(N-methylacetamide)adenosine-3{grave over ( )}-phosphorothioate 
               
               
                 (Gam) 
                 2{grave over ( )}-O-(N-methylacetamide)guanosine-3{grave over ( )}-phosphate 
               
               
                 (Gams) 
                 2{grave over ( )}-O-(N-methylacetamide)guanosine-3{grave over ( )}-phosphorothioate 
               
               
                 (Tam) 
                 2{grave over ( )}-O-(N-methylacetamide)thymidine-3{grave over ( )}-phosphate 
               
               
                 (Tams) 
                 2{grave over ( )}-O-(N-methylacetamide)thymidine-3{grave over ( )}-phosphorothioate 
               
               
                 dA 
                 2{grave over ( )}-deoxyadenosine-3{grave over ( )}-phosphate 
               
               
                 dAs 
                 2{grave over ( )}-deoxyadenosine-3{grave over ( )}-phosphorothioate 
               
               
                 dC 
                 2{grave over ( )}-deoxycytidine-3{grave over ( )}-phosphate 
               
               
                 dCs 
                 2{grave over ( )}-deoxycytidine-3{grave over ( )}-phosphorothioate 
               
               
                 dG 
                 2{grave over ( )}-deoxyguanosine-3{grave over ( )}-phosphate 
               
               
                 dGs 
                 2{grave over ( )}-deoxyguanosine-3{grave over ( )}-phosphorothioate 
               
               
                 dT 
                 2{grave over ( )}-deoxythymidine-3{grave over ( )}-phosphate 
               
               
                 dTs 
                 2{grave over ( )}-deoxythymidine-3{grave over ( )}-phosphorothioate 
               
               
                 dU 
                 2{grave over ( )}-deoxyuridine 
               
               
                 dUs 
                 2{grave over ( )}-deoxyuridine-3{grave over ( )}-phosphorothioate 
               
               
                 (Aeo) 
                 2{grave over ( )}-O-methoxyethyladenosine-3{grave over ( )}-phosphate 
               
               
                 (Aeos) 
                 2{grave over ( )}-O-methoxyethyladenosine-3{grave over ( )}-phosphorothioate 
               
               
                 (Geo) 
                 2{grave over ( )}-O-methoxyethylguanosine-3{grave over ( )}-phosphate 
               
               
                 (Geos) 
                 2{grave over ( )}-O-methoxyethylguanosine-3{grave over ( )}-phosphorothioate 
               
               
                 (Teo) 
                 2{grave over ( )}-O-methoxyethyl-5-methyluridine-3{grave over ( )}-phosphate 
               
               
                 (Teos) 
                 2{grave over ( )}-O-methoxyethyl-5-methyluridine-3{grave over ( )}-phosphorothioate 
               
               
                 (m5Ceo) 
                 2{grave over ( )}-O-methoxyethyl-5-methylcytidine-3{grave over ( )}-phosphate 
               
               
                 (m5Ceos) 
                 2{grave over ( )}-O-methoxyethyl-5-methylcytidine-3{grave over ( )}-phosphorothioate 
               
               
                 (A3m) 
                 3{grave over ( )}-O-methyladenosine-2{grave over ( )}-phosphate 
               
               
                 (A3mx) 
                 3{grave over ( )}-O-methyl-xylofuranosyladenosine-2{grave over ( )}-phosphate 
               
               
                 (G3m) 
                 3{grave over ( )}-O-methylguanosine-2{grave over ( )}-phosphate 
               
               
                 (G3mx) 
                 3{grave over ( )}-O-methyl-xylofuranosylguanosine-2{grave over ( )}-phosphate 
               
               
                 (C3m) 
                 3{grave over ( )}-O-methylcytidine-2{grave over ( )}-phosphate 
               
               
                 (C3mx) 
                 3{grave over ( )}-O-methyl-xylofuranosylcytidine-2{grave over ( )}-phosphate 
               
               
                 (U3m) 
                 3{grave over ( )}-O-methyluridine-2{grave over ( )}-phosphate 
               
               
                 U3mx) 
                 3{grave over ( )}-O-methyl-xylofuranosyluridine-2{grave over ( )}-phosphate 
               
               
                 (m5Cam) 
                 2{grave over ( )}-O-(N-methylacetamide)-5-methylcytidine-3{grave over ( )}-phosphate 
               
               
                 (m5Cams) 
                 2{grave over ( )}-O-(N-methylacetamide)-5-methylcytidine- 
               
               
                   
                 3{grave over ( )}-phosphorothioate 
               
               
                 (Chd) 
                 2{grave over ( )}-O-hexadecyl-cytidine-3{grave over ( )}-phosphate 
               
               
                 (Chds) 
                 2{grave over ( )}-O-hexadecyl-cytidine-3{grave over ( )}-phosphorothioate 
               
               
                 (Uhd) 
                 2{grave over ( )}-O-hexadecyl-uridine-3{grave over ( )}-phosphate 
               
               
                 (Uhds) 
                 2{grave over ( )}-O-hexadecyl-uridine-3{grave over ( )}-phosphorothioate 
               
               
                 (pshe) 
                 Hydroxyethylphosphorothioate 
               
               
                   
               
            
           
         
       
     
     
       
         
           
               
             
               
                 TABLE 2 
               
             
            
               
                   
               
               
                  Unmodified Sense and Antisense Strand Sequences of SERPINF2 dsRNA Agents 
               
            
           
           
               
               
               
               
               
               
               
               
               
            
               
                   
                   
                 Unmodified  
                   
                   
                   
                   
                   
                   
               
               
                   
                 Sense  
                 Sense Sequence 
                   
                   
                 Antisense 
                 Unmodified antisense Sequence 
                   
                   
               
               
                 Duplex Name 
                 Oligo Name 
                 5′ to 3′  
                 SEQ ID NO: 
                 Source 
                 Oligo Name 
                 5′ to 3′  
                 SEQ ID NO: 
                 Source 
               
               
                   
               
               
                 AD-203163.1 
                 A-401843.1 
                 UGUAAGGUUUUUGUAG 
                  29 
                 NM__000934.3_2233- 
                 A-401844.1 
                 AAUCACUACAAAAACCUUACAAA 
                 119 
                 NM__000934.3_2231- 
               
               
                   
                   
                 UGAUU 
                   
                 2253_s 
                   
                   
                   
                 2253_as 
               
               
                   
               
               
                 AD-203147.1 
                 A-401811.1 
                 GCUCCUUAAGGCUCUU 
                  30 
                 NM__000934.3_2216- 
                 A-401812.1 
                 UACAAAAGAGCCUUAAGGAGCUC 
                 120 
                 NM__000934.3_2214- 
               
               
                   
                   
                 UUGUA 
                   
                 2236_s 
                   
                   
                   
                 2236_as 
               
               
                   
               
               
                 AD-202129.1 
                 A-399775.1 
                 ACCUGGCAAACAUCAA 
                  31 
                 NM__000934.3_676- 
                 A-399776.1 
                 AUUGGUUGAUGUUUGCCAGGUCA 
                 121 
                 NM__000934.3_674- 
               
               
                   
                   
                 CCAAU 
                   
                 696_s 
                   
                   
                   
                 696_as 
               
               
                   
               
               
                 AD-203171.1 
                 A-401859.1 
                 UAGUGAUUUUUAUGCC 
                  32 
                 NM__000934.3_2246- 
                 A-401860.1 
                 UAGGUGGCAUAAAAAUCACUACA 
                 122 
                 NM__000934.3_2244- 
               
               
                   
                   
                 ACCUA 
                   
                 2266_G21A_s 
                   
                   
                   
                 2266_C1U_as 
               
               
                   
               
               
                 AD-202239.1 
                 A-399995.1 
                 AACAAGUUUGACCCGA 
                  33 
                 NM__000934.3_804- 
                 A-399996.1 
                 AAGGCUCGGGUCAAACUUGUUCC 
                 123 
                 NM__000934.3_802- 
               
               
                   
                   
                 GCCUU 
                   
                 824_s 
                   
                   
                   
                 824_as 
               
               
                   
               
               
                 AD-202137.1 
                 A-399791.1 
                 AACAUCAACCAAUGGG 
                  34 
                 NM_000934.3_684- 
                 A-399792.1 
                 UUUCACCCAUUGGUUGAUGUUUG 
                 124 
                 NM__000934.3_682- 
               
               
                   
                   
                 UGAAA 
                   
                 704_G21A_s 
                   
                   
                   
                 704_C1U_as 
               
               
                   
               
               
                 AD-202196.1 
                 A-399909.1 
                 GUUGCUUCUCCUCAAC 
                  35 
                 NM__000934.3_761- 
                 A-399910.1 
                 AUGGCGUUGAGGAGAAGCAACAC 
                 125 
                 NM__000934.3_759- 
               
               
                   
                   
                 GCCAU 
                   
                 781_s 
                   
                   
                   
                 781_as 
               
               
                   
               
               
                 AD-203162.1 
                 A-401841.1 
                 UUGUAAGGUUUUUGUA 
                  36 
                 NM__000934.3_2232- 
                 A-401842.1 
                 AUCACUACAAAAACCUUACAAAA 
                 126 
                 NM__000934.3_2230- 
               
               
                   
                   
                 GUGAU 
                   
                 2252_s 
                   
                   
                   
                 2252_as 
               
               
                   
               
               
                 AD-202130.1 
                 A-399777.1 
                 CCUGGCAAACAUCAAC 
                  37 
                 NM__000934.3_677- 
                 A-399778.1 
                 UAUUGGUUGAUGUUUGCCAGGUC 
                 127 
                 NM__000934.3_675- 
               
               
                   
                   
                 CAAUA 
                   
                 697_G21A_s 
                   
                   
                   
                 697_C1U_as 
               
               
                   
               
               
                 AD-203169.1 
                 A-401855.1 
                 UGUAGUGAUUUUUAUG 
                  38 
                 NM__000934.3_2244- 
                 A-401856.1 
                 UGUGGCAUAAAAAUCACUACAAA 
                 128 
                 NM__000934.3_2242- 
               
               
                   
                   
                 CCACA 
                   
                 2264_C21A_s 
                   
                   
                   
                 2264_G1U_as 
               
               
                   
               
               
                 AD-202052.1 
                 A-399621.1 
                 UGCAGAAAGGAUUUCC 
                  39 
                 NM__000934.3_580- 
                 A-399622.1 
                 UGAUGGGAAAUCCUUUCUGCAGG 
                 129 
                 NM__000934.3_578- 
               
               
                   
                   
                 CAUCA 
                   
                 600_s 
                   
                   
                   
                 600_as 
               
               
                   
               
               
                 AD-203184.1 
                 A-401885.1 
                 GCCACCUGAAUAAAGA 
                  40 
                 NM__000934.3_2259- 
                 A-401886.1 
                 UUCAUUCUUUAUUCAGGUGGCAU 
                 130 
                 NM__000934.3_2257- 
               
               
                   
                   
                 AUGAA 
                   
                 2279_s 
                   
                   
                   
                 2279_as 
               
               
                   
               
               
                 AD-202492.1 
                 A-400501.1 
                 CCUAAGCUGUAUCUGA 
                  41 
                 NM__000934.3_1086- 
                 A-400502.1 
                 UUGUUUCAGAUACAGCUUAGGCA 
                 131 
                 NM__000934.3_1084- 
               
               
                   
                   
                 AACAA 
                   
                 1106_C21A_s 
                   
                   
                   
                 1106_G1U_as 
               
               
                   
               
               
                 AD-202491.1 
                 A-400499.1 
                 GCCUAAGCUGUAUCUG 
                  42 
                 NM__000934.3_1085- 
                 A-400500.1 
                 UGUUUCAGAUACAGCUUAGGCAG 
                 132 
                 NM__000934.3_1083- 
               
               
                   
                   
                 AAACA 
                   
                 1105_s 
                   
                   
                   
                 1105_as 
               
               
                   
               
               
                 AD-202050.1 
                 A-399617.1 
                 CCUGCAGAAAGGAUUU 
                  43 
                 NM__000934.3_578- 
                 A-399618.1 
                 AUGGGAAAUCCUUUCUGCAGGUA 
                 133 
                 NM__000934.3_576- 
               
               
                   
                   
                 CCCAU 
                   
                 598_s 
                   
                   
                   
                 598_as 
               
               
                   
               
               
                 AD-202197.1 
                 A-399911.1 
                 UUGCUUCUCCUCAACG 
                  44 
                 NM__000934.3_762- 
                 A-399912.1 
                 UAUGGCGUUGAGGAGAAGCAACA 
                 134 
                 NM__000934.3_760- 
               
               
                   
                   
                 CCAUA 
                   
                 782_C21A_s 
                   
                   
                   
                 782_G1U_as 
               
               
                   
               
               
                 AD-202192.1 
                 A-399901.1 
                 CCGUGUUGCUUCUCCU 
                  45 
                 NM__000934.3_757- 
                 A-399902.1 
                 UGUUGAGGAGAAGCAACACGGUG 
                 135 
                 NM__000934.3_755- 
               
               
                   
                   
                 CAACA 
                   
                 777_G21A_s 
                   
                   
                   
                 777_C1U_as 
               
               
                   
               
               
                 AD-203148.1 
                 A-401813.1 
                 CUCCUUAAGGCUCUUU 
                  46 
                 NM__000934.3_2217- 
                 A-401814.1 
                 UUACAAAAGAGCCUUAAGGAGCU 
                 136 
                 NM__000934.3_2215- 
               
               
                   
                   
                 UGUAA 
                   
                 2237_s 
                   
                   
                   
                 2237_as 
               
               
                   
               
               
                 AD-202499.1 
                 A-400515.1 
                 UGUAUCUGAAACACCA 
                  47 
                 NM__000934.3_1093- 
                 A-400516.1 
                 UCAUUUGGUGUUUCAGAUACAGC 
                 137 
                 NM__000934.3_1091- 
               
               
                   
                   
                 AAUGA 
                   
                 1113_G21A_s 
                   
                   
                   
                 1113_C1U_as 
               
               
                   
               
               
                 AD-202054.1 
                 A-399625.1 
                 CAGAAAGGAUUUCCCA 
                  48 
                 NM__000934.3_582- 
                 A-399626.1 
                 UUUGAUGGGAAAUCCUUUCUGCA 
                 138 
                 NM__000934.3_580- 
               
               
                   
                   
                 UCAAA 
                   
                 602_s 
                   
                   
                   
                 602_as 
               
               
                   
               
               
                 AD-202372.1 
                 A-400261.1 
                 GAGCUUUGUGGUCCUU 
                  49 
                 NM__000934.3_965- 
                 A-400262.1 
                 UGUACAAGGACCACAAAGCUCAU 
                 139 
                 NM__000934.3_963- 
               
               
                   
                   
                 GUACA 
                   
                 985_C21A_s 
                   
                   
                   
                 985_G1U_as 
               
               
                   
               
               
                 AD-202796.1 
                 A-401109.1 
                 CUCAUGUGACUCUUUC 
                  50 
                 NM__000934.3_1603- 
                 A-401110.1 
                 UGUUGGAAAGAGUCACAUGAGUG 
                 140 
                 NM__000934.3_1601- 
               
               
                   
                   
                 CAACA 
                   
                 1623_C21A_s 
                   
                   
                   
                 1623_G1U_as 
               
               
                   
               
               
                 AD-206288.1 
                 A-408120.1 
                 GCUCCCAAGACUCUUU 
                  51 
                 NM__008878.2_2125- 
                 A-408121.1 
                 UUACAAAAGAGUCUUGGGAGCUA 
                 141 
                 NM__008878.2_2123- 
               
               
                   
                   
                 UGUAA 
                   
                 2145_s 
                   
                   
                   
                 2145_as 
               
               
                   
               
               
                 AD-202191.1 
                 A-399899.1 
                 ACCGUGUUGCUUCUCC 
                  52 
                 NM_000934.3_756- 
                 A-399900.1 
                 UUUGAGGAGAAGCAACACGGUGU 
                 142 
                 NM__000934.3_754- 
               
               
                   
                   
                 UCAAA 
                   
                 776_C21A_s 
                   
                   
                   
                 776_G1U_as 
               
               
                   
               
               
                 AD-203177.1 
                 A-401871.1 
                 UUUUUAUGCCACCUGA 
                  53 
                 NM__000934.3_2252- 
                 A-401872.1 
                 UUUAUUCAGGUGGCAUAAAAAUC 
                 143 
                 NM__000934.3_2250- 
               
               
                   
                   
                 AUAAA 
                   
                 2272_s 
                   
                   
                   
                 2272_as 
               
               
                   
               
               
                 AD-202489.1 
                 A-400495.1 
                 CUGCCUAAGCUGUAUC 
                  54 
                 NM__000934.3_1083- 
                 A-400496.1 
                 UUUCAGAUACAGCUUAGGCAGCC 
                 144 
                 NM__000934.3_1081- 
               
               
                   
                   
                 UGAAA 
                   
                 1103_s 
                   
                   
                   
                 1103_as 
               
               
                   
               
               
                 AD-202387.1 
                 A-400291.1 
                 UGUACCCACCCACUUU 
                  55 
                 NM__000934.3_980- 
                 A-400292.1 
                 UAUUCAAAGUGGGUGGGUACAAG 
                 145 
                 NM__000934.3_978- 
               
               
                   
                   
                 GAAUA 
                   
                 1000_G21A_s 
                   
                   
                   
                 1000_C1U_as 
               
               
                   
               
               
                 AD-202399.1 
                 A-400315.1 
                 CUUUGAAUGGAACGUG 
                  56 
                 NM__000934.3_992- 
                 A-400316.1 
                 UGGGACACGUUCCAUUCAAAGUG 
                 146 
                 NM__000934.3_990- 
               
               
                   
                   
                 UCCCA 
                   
                 1012_s 
                   
                   
                   
                 1012_as 
               
               
                   
               
               
                 AD-202386.1 
                 A-400289.1 
                 UUGUACCCACCCACUU 
                  57 
                 NM__000934.3_979- 
                 A-400290.1 
                 AUUCAAAGUGGGUGGGUACAAGG 
                 147 
                 NM__000934.3_977- 
               
               
                   
                   
                 UGAAU 
                   
                 999_s 
                   
                   
                   
                 999_as 
               
               
                   
               
               
                 AD-202053.1 
                 A-399623.1 
                 GCAGAAAGGAUUUCCC 
                  58 
                 NM__000934.3_581- 
                 A-399624.1 
                 UUGAUGGGAAAUCCUUUCUGCAG 
                 148 
                 NM__000934.3_579- 
               
               
                   
                   
                 AUCAA 
                   
                 601_s 
                   
                   
                   
                 601_as 
               
               
                   
               
               
                 AD-203166.1 
                 A-401849.1 
                 UUUUGUAGUGAUUUUU 
                  59 
                 NM__000934.3_2241- 
                 A-401850.1 
                 UGCAUAAAAAUCACUACAAAAAC 
                 149 
                 NM__000934.3_2239- 
               
               
                   
                   
                 AUGCA 
                   
                 2261_C21A_s 
                   
                   
                   
                 2261_G1U_as 
               
               
                   
               
               
                 AD-202827.1 
                 A-401171.1 
                 GAGGCCAUUCUUUCCC 
                  60 
                 NM__000934.3_1665- 
                 A-401172.1 
                 UUGUUGGGAAAGAAUGGCCUCUC 
                 150 
                 NM__000934.3_1663- 
               
               
                   
                   
                 AACAA 
                   
                 1685_C21A_s 
                   
                   
                   
                 1685_G1U_as 
               
               
                   
               
               
                 AD-205123.1 
                 A-405796.1 
                 GAUUUCCUGGAGCAAU 
                  61 
                 NM__008878.2_578- 
                 A-405797.1 
                 UUCCGAUUGCUCCAGGAAAUCGU 
                 151 
                 NM__008878.2_576- 
               
               
                   
                   
                 CGGAA 
                   
                 598_s 
                   
                   
                   
                 598_as 
               
               
                   
               
               
                 AD-202490.1 
                 A-400497.1 
                 UGCCUAAGCUGUAUCU 
                  62 
                 NM__000934.3_1084- 
                 A-400498.1 
                 GUUUCAGAUACAGCUUAGGCAGC 
                 152 
                 NM__000934.3_1082- 
               
               
                   
                   
                 GAAAC 
                   
                 1104_s 
                   
                   
                   
                 1104_as 
               
               
                   
               
               
                 AD-202051.1 
                 A-399619.1 
                 CUGCAGAAAGGAUUUC 
                  63 
                 NM__000934.3_579- 
                 A-399620.1 
                 UAUGGGAAAUCCUUUCUGCAGGU 
                 153 
                 NM__000934.3_577- 
               
               
                   
                   
                 CCAUA 
                   
                 599_C21A_s 
                   
                   
                   
                 599_G1U_as 
               
               
                   
               
               
                 AD-202894.1 
                 A-401305.1 
                 AGGGAGGUGCUUCUAG 
                  64 
                 NM__000934.3_1815- 
                 A-401306.1 
                 UAGAACUAGAAGCACCUCCCUCC 
                 154 
                 NM__000934.3_1813- 
               
               
                   
                   
                 UUCUA 
                   
                 1835_G21A_s 
                   
                   
                   
                 1835_C1U_as 
               
               
                   
               
               
                 AD-202494.1 
                 A-400505.1 
                 UAAGCUGUAUCUGAAA 
                  65 
                 NM__000934.3_1088- 
                 A-400506.1 
                 UGGUGUUUCAGAUACAGCUUAGG 
                 155 
                 NM__000934.3_1086- 
               
               
                   
                   
                 CACCA 
                   
                 1108_s 
                   
                   
                   
                 1108_as 
               
               
                   
               
               
                 AD-202905.1 
                 A-401327.1 
                 UCUAGUUCUGCCAGGA 
                  66 
                 NM__000934.3_1826- 
                 A-401328.1 
                 UUGUCUCCUGGCAGAACUAGAAG 
                 156 
                 NM__000934.3_1824- 
               
               
                   
                   
                 GACAA 
                   
                 1846_G21A_s 
                   
                   
                   
                 1846_C1U_as 
               
               
                   
               
               
                 AD-202904.1 
                 A-401325.1 
                 UUCUAGUUCUGCCAGG 
                  67 
                 NM_000934.3_1825- 
                 A-401326.1 
                 UGUCUCCUGGCAGAACUAGAAGC 
                 157 
                 NM_000934.3_1823- 
               
               
                   
                   
                 AGACA 
                   
                 1845_s 
                   
                   
                   
                 1845_as 
               
               
                   
               
               
                 AD-203151.1 
                 A-401819.1 
                 CUUAAGGCUCUUUUGU 
                  68 
                 NM_000934.3_2220- 
                 A-401820.1 
                 ACCUUACAAAAGAGCCUUAAGGA 
                 158 
                 NM_000934.3_2218- 
               
               
                   
                   
                 AAGGU 
                   
                 2240_s 
                   
                   
                   
                 2240_as 
               
               
                   
               
               
                 AD-202406.1 
                 A-400329.1 
                 UGGAACGUGUCCCAGG 
                  69 
                 NM_000934.3_999- 
                 A-400330.1 
                 UAGUACCUGGGACACGUUCCAUU 
                 159 
                 NM_000934.3_997- 
               
               
                   
                   
                 UACUA 
                   
                 1019_G21A_s 
                   
                   
                   
                 1019_C1U_as 
               
               
                   
               
               
                 AD-202913.1 
                 A-401343.1 
                 UGCCAGGAGACAGGUU 
                  70 
                 NM_000934.3_1834- 
                 A-401344.1 
                 UAGCUAACCUGUCUCCUGGCAGA 
                 160 
                 NM__000934.3_1832- 
               
               
                   
                   
                 AGCUA 
                   
                 1854_G21A_s 
                   
                   
                   
                 1854_C1U_as 
               
               
                   
               
               
                 AD-202277.1 
                 A-400071.1 
                 CCACCUGGACGAGCAG 
                  71 
                 NM_000934.3_842- 
                 A-400072.1 
                 UUGAACUGCUCGUCCAGGUGGAA 
                 161 
                 NM_000934.3_840- 
               
               
                   
                   
                 UUCAA 
                   
                 862_C21A_s 
                   
                   
                   
                 862_G1U_as 
               
               
                   
               
               
                 AD-202133.1 
                 A-399783.1 
                 GGCAAACAUCAACCAA 
                  72 
                 NM__000934.3_680- 
                 A-399784.1 
                 ACCCAUUGGUUGAUGUUUGCCAG 
                 162 
                 NM__000934.3_678- 
               
               
                   
                   
                 UGGGU 
                   
                 700_s 
                   
                   
                   
                 700_as 
               
               
                   
               
               
                 AD-202895.1 
                 A-401307.1 
                 GGGAGGUGCUUCUAGU 
                  73 
                 NM_000934.3_1816- 
                 A-401308.1 
                 UCAGAACUAGAAGCACCUCCCUC 
                 163 
                 NM_000934.3_1814- 
               
               
                   
                   
                 UCUGA 
                   
                 1836_C21A_s 
                   
                   
                   
                 1836_G1U_as 
               
               
                   
               
               
                 AD-202398.1 
                 A-400313.1 
                 ACUUUGAAUGGAACGU 
                  74 
                 NM__000934.3_991- 
                 A-400314.1 
                 UGGACACGUUCCAUUCAAAGUGG 
                 164 
                 NM__000934.3_989- 
               
               
                   
                   
                 GUCCA 
                   
                 1011_C21A_s 
                   
                   
                   
                 1011_G1U_as 
               
               
                   
               
               
                 AD-202136.1 
                 A-399789.1 
                 AAACAUCAACCAAUGG 
                  75 
                 NM__000934.3_683- 
                 A-399790.1 
                 UUCACCCAUUGGUUGAUGUUUGC 
                 165 
                 NM__000934.3_681- 
               
               
                   
                   
                 GUGAA 
                   
                 703_s 
                   
                   
                   
                 703_as 
               
               
                   
               
               
                 AD-202131.1 
                 A-399779.1 
                 CUGGCAAACAUCAACC 
                  76 
                 NM_000934.3_678- 
                 A-399780.1 
                 UCAUUGGUUGAUGUUUGCCAGGU 
                 166 
                 NM_000934.3_676- 
               
               
                   
                   
                 AAUGA 
                   
                 698_G21A_s 
                   
                   
                   
                 698_C1U_as 
               
               
                   
               
               
                 AD-202393.1 
                 A-400303.1 
                 CACCCACUUUGAAUGG 
                  77 
                 NM_000934.3_986- 
                 A-400304.1 
                 ACGUUCCAUUCAAAGUGGGUGGG 
                 167 
                 NM_000934.3_984- 
               
               
                   
                   
                 AACGU 
                   
                 1006_s 
                   
                   
                   
                 1006_as 
               
               
                   
               
               
                 AD-201919.1 
                 A-399355.1 
                 CUCCCUGGUGGCUCAA 
                  78 
                 NM_000934.3_359- 
                 A-399356.1 
                 UACGUUUGAGCCACCAGGGAGAA 
                 168 
                 NM_000934.3_357- 
               
               
                   
                   
                 ACGUA 
                   
                 379_C21A_s 
                   
                   
                   
                 379_G1U_as 
               
               
                   
               
               
                 AD-202787.1 
                 A-401091.1 
                 GCCUCUCCACUCAUGU 
                  79 
                 NM__000934.3_1594- 
                 A-401092.1 
                 UAGUCACAUGAGUGGAGAGGCUG 
                 169 
                 NM__000934.3_1592- 
               
               
                   
                   
                 GACUA 
                   
                 1614_C21A_s 
                   
                   
                   
                 1614_G1U_as 
               
               
                   
               
               
                 AD-205414.1 
                 A-406374.1 
                 UGAGAUACAGGUGGCU 
                  80 
                 NM__008878.2_898- 
                 A-406375.1 
                 AAAUGAGCCACCUGUAUCUCAGG 
                 170 
                 NM__008878.2_896- 
               
               
                   
                   
                 CAUUU 
                   
                 918_s 
                   
                   
                   
                 918_as 
               
               
                   
               
               
                 AD-205279.1 
                 A-406104.1 
                 CGGUUUCUGGAGGACC 
                  81 
                 NM__008878.2_763- 
                 A-406105.1 
                 AACUUGGUCCUCCAGAAACCGUG 
                 171 
                 NM__008878.2_761- 
               
               
                   
                   
                 AAGUU 
                   
                 783_s 
                   
                   
                   
                 783_as 
               
               
                   
               
               
                 AD-202373.1 
                 A-400263.1 
                 AGCUUUGUGGUCCUUG 
                  82 
                 NM_000934.3_966- 
                 A-400264.1 
                 UGGUACAAGGACCACAAAGCUCA 
                 172 
                 NM_000934.3_964- 
               
               
                   
                   
                 UACCA 
                   
                 986_C21A_s 
                   
                   
                   
                 986_G1U_as 
               
               
                   
               
               
                 AD-202198.1 
                 A-399913.1 
                 UGCUUCUCCUCAACGC 
                  83 
                 NM_000934.3_763- 
                 A-399914.1 
                 UGAUGGCGUUGAGGAGAAGCAAC 
                 173 
                 NM_000934.3_761- 
               
               
                   
                   
                 CAUCA 
                   
                 783_C21A_s 
                   
                   
                   
                 783_G1U_as 
               
               
                   
               
               
                 AD-202420.1 
                 A-400357.1 
                 GGUACUGGCCAACCUG 
                  84 
                 NM_000934.3_1013- 
                 A-400358.1 
                 UAACUCAGGUUGGCCAGUACCUG 
                 174 
                 NM_000934.3_1011- 
               
               
                   
                   
                 AGUUA 
                   
                 1033_G21A_s 
                   
                   
                   
                 1033_C1U_as 
               
               
                   
               
               
                 AD-202199.1 
                 A-399915.1 
                 GCUUCUCCUCAACGCC 
                  85 
                 NM__000934.3_764- 
                 A-399916.1 
                 UGGAUGGCGUUGAGGAGAAGCAA 
                 175 
                 NM__000934.3_762- 
               
               
                   
                   
                 AUCCA 
                   
                 784_s 
                   
                   
                   
                 784_as 
               
               
                   
               
               
                 AD-204885.1 
                 A-405324.1 
                 CAUGAUGGCUUUUACU 
                  86 
                 NM__008878.2_304- 
                 A-405325.1 
                 UCAGUAGUAAAAGCCAUCAUGGC 
                 176 
                 NM__008878.2_302- 
               
               
                   
                   
                 ACUGA 
                   
                 324_s 
                   
                   
                   
                 324_as 
               
               
                   
               
               
                 AD-205284.1 
                 A-406114.1 
                 UCUGGAGGACCAAGUU 
                  87 
                 NM__008878.2_768- 
                 A-406115.1 
                 UGUCAAACUUGGUCCUCCAGAAA 
                 177 
                 NM__008878.2_766- 
               
               
                   
                   
                 UGACA 
                   
                 788_C21A_s 
                   
                   
                   
                 788_G1U_as 
               
               
                   
               
               
                 AD-205458.1 
                 A-406462.1 
                 UGGAACGUGUCCGAGG 
                  88 
                 NM__008878.2_971- 
                 A-406463.1 
                 UAGUACCUCGGACACGUUCCACU 
                 178 
                 NM__008878.2_969- 
               
               
                   
                   
                 UACUA 
                   
                 991_s 
                   
                   
                   
                 991_as 
               
               
                   
               
               
                 AD-205413.1 
                 A-406372.1 
                 CUGAGAUACAGGUGGC 
                  89 
                 NM__008878.2_897- 
                 A-406373.1 
                 AAUGAGCCACCUGUAUCUCAGGU 
                 179 
                 NM__008878.2_895- 
               
               
                   
                   
                 UCAUU 
                   
                 917_s 
                   
                   
                   
                 917_as 
               
               
                   
               
               
                 AD-206058.1 
                 A-407660.1 
                 GUCCCUCCACAUUUGU 
                  90 
                 NM__008878.2_1804- 
                 A-407661.1 
                 UGCUUACAAAUGUGGAGGGACCC 
                 180 
                 NM__008878.2_1802- 
               
               
                   
                   
                 AAGCA 
                   
                 1824_s 
                   
                   
                   
                 1824_as 
               
               
                   
               
               
                 AD-205411.1 
                 A-406368.1 
                 ACCUGAGAUACAGGUG 
                  91 
                 NM__008878.2_895- 
                 A-406369.1 
                 UGAGCCACCUGUAUCUCAGGUUG 
                 181 
                 NM__008878.2_893- 
               
               
                   
                   
                 GCUCA 
                   
                 915_s 
                   
                   
                   
                 915_as 
               
               
                   
               
               
                 AD-202407.1 
                 A-400331.1 
                 GGAACGUGUCCCAGGU 
                  92 
                 NM__000934.3_1000- 
                 A-400332.1 
                 UCAGUACCUGGGACACGUUCCAU 
                 182 
                 NM__000934.3_998- 
               
               
                   
                   
                 ACUGA 
                   
                 1020_G21A_s 
                   
                   
                   
                 1020_C1U_as 
               
               
                   
               
               
                 AD-205459.1 
                 A-406464.1 
                 GGAACGUGUCCGAGGU 
                  93 
                 NM__008878.2_972- 
                 A-406465.1 
                 UUAGUACCUCGGACACGUUCCAC 
                 183 
                 NM__008878.2_970- 
               
               
                   
                   
                 ACUAA 
                   
                 992_G21A_s 
                   
                   
                   
                 992_C1U_as 
               
               
                   
               
               
                 AD-205455.1 
                 A-406456.1 
                 GAGUGGAACGUGUCCG 
                  94 
                 NM__008878.2_968- 
                 A-406457.1 
                 UACCUCGGACACGUUCCACUCAA 
                 184 
                 NM__008878.2_966- 
               
               
                   
                   
                 AGGUA 
                   
                 988_s 
                   
                   
                   
                 988_as 
               
               
                   
               
               
                 AD-205264.1 
                 A-406074.1 
                 CGCCAUCCACUUUCAC 
                  95 
                 NM__008878.2_748- 
                 A-406075.1 
                 AAACCGUGAAAGUGGAUGGCGUU 
                 185 
                 NM__008878.2_746- 
               
               
                   
                   
                 GGUUU 
                   
                 768_s 
                   
                   
                   
                 768_as 
               
               
                   
               
               
                 AD-204884.1 
                 A-405322.1 
                 CCAUGAUGGCUUUUAC 
                  96 
                 NM__008878.2_303- 
                 A-405323.1 
                 UAGUAGUAAAAGCCAUCAUGGCC 
                 186 
                 NM__008878.2_301- 
               
               
                   
                   
                 UACUA 
                   
                 323_G21A_s 
                   
                   
                   
                 323_C1U_as 
               
               
                   
               
               
                 AD-204888.1 
                 A-405330.1 
                 GAUGGCUUUUACUACU 
                  97 
                 NM__008878.2_307- 
                 A-405331.1 
                 AGGUCAGUAGUAAAAGCCAUCAU 
                 187 
                 NM__008878.2_305- 
               
               
                   
                   
                 GACCU 
                   
                 327_s 
                   
                   
                   
                 327_as 
               
               
                   
               
               
                 AD-205270.1 
                 A-406086.1 
                 CCACUUUCACGGUUUC 
                  98 
                 NM__008878.2_754- 
                 A-406087.1 
                 UUCCAGAAACCGUGAAAGUGGAU 
                 188 
                 NM__008878.2_752- 
               
               
                   
                   
                 UGGAA 
                   
                 774_G21A_s 
                   
                   
                   
                 774_C1U_as 
               
               
                   
               
               
                 AD-204894.1 
                 A-405342.1 
                 UUUUACUACUGACCUG 
                  99 
                 NM__008878.2_313- 
                 A-405343.1 
                 UAGAACAGGUCAGUAGUAAAAGC 
                 189 
                 NM__008878.2_311- 
               
               
                   
                   
                 UUCUA 
                   
                 333_C21A_s 
                   
                   
                   
                 333_G1U_as 
               
               
                   
               
               
                 AD-205273.1 
                 A-406092.1 
                 CUUUCACGGUUUCUGG 
                 100 
                 NM__008878.2_757- 
                 A-406093.1 
                 UUCCUCCAGAAACCGUGAAAGUG 
                 190 
                 NM__008878.2_755- 
               
               
                   
                   
                 AGGAA 
                   
                 777_C21A_s 
                   
                   
                   
                 777_G1U_as 
               
               
                   
               
               
                 AD-206218.1 
                 A-407980.1 
                 UUGCUGAAGACAUUCA 
                 101 
                 NM__008878.2_1966- 
                 A-407981.1 
                 UGACCUGAAUGUCUUCAGCAAGG 
                 191 
                 NM__008878.2_1964- 
               
               
                   
                   
                 GGUCA 
                   
                 1986_C21A_s 
                   
                   
                   
                 1986_G1U_as 
               
               
                   
               
               
                 AD-202132.1 
                 A-399781.1 
                 UGGCAAACAUCAACCA 
                 102 
                 NM__000934.3_679- 
                 A-399782.1 
                 UCCAUUGGUUGAUGUUUGCCAGG 
                 192 
                 NM__000934.3_677- 
               
               
                   
                   
                 AUGGA 
                   
                 699_G21A_s 
                   
                   
                   
                 699_C1U_as 
               
               
                   
               
               
                 AD-202389.1 
                 A-400295.1 
                 UACCCACCCACUUUGA 
                 103 
                 NM__000934.3_982- 
                 A-400296.1 
                 UCCAUUCAAAGUGGGUGGGUACA 
                 193 
                 NM__000934.3_980- 
               
               
                   
                   
                 AUGGA 
                   
                 1002_s 
                   
                   
                   
                 1002_as 
               
               
                   
               
               
                 AD-204944.1 
                 A-405442.1 
                 AGUGUGGCCCUAGCAC 
                 104 
                 NM__008878.2_383- 
                 A-405443.1 
                 AGAGAGUGCUAGGGCCACACUAA 
                 194 
                 NM__008878.2_381- 
               
               
                   
                   
                 UCUCU 
                   
                 403_s 
                   
                   
                   
                 403_as 
               
               
                   
               
               
                 AD-205463.1 
                 A-406472.1 
                 CGUGUCCGAGGUACUA 
                 105 
                 NM__008878.2_976- 
                 A-406473.1 
                 UUGGCUAGUACCUCGGACACGUU 
                 195 
                 NM__008878.2_974- 
               
               
                   
                   
                 GCCAA 
                   
                 996_s 
                   
                   
                   
                 996_as 
               
               
                   
               
               
                 AD-206057.1 
                 A-407658.1 
                 GGUCCCUCCACAUUUG 
                 106 
                 NM__008878.2_1803- 
                 A-407659.1 
                 UCUUACAAAUGUGGAGGGACCCU 
                 196 
                 NM__008878.2_1801- 
               
               
                   
                   
                 UAAGA 
                   
                 1823_C21A_s 
                   
                   
                   
                 1823_G1U_as 
               
               
                   
               
               
                 AD-206281.1 
                 A-408106.1 
                 CCCUUUAGCUCCCAAG 
                 107 
                 NM__008878.2_2118- 
                 A-408107.1 
                 AGAGUCUUGGGAGCUAAAGGGUC 
                 197 
                 NM__008878.2_2116- 
               
               
                   
                   
                 ACUCU 
                   
                 2138_s 
                   
                   
                   
                 2138_as 
               
               
                   
               
               
                 AD-206059.1 
                 A-407662.1 
                 UCCCUCCACAUUUGUA 
                 108 
                 NM__008878.2_1805- 
                 A-407663.1 
                 UUGCUUACAAAUGUGGAGGGACC 
                 198 
                 NM__008878.2_1803- 
               
               
                   
                   
                 AGCAA 
                   
                 1825_G21A_s 
                   
                   
                   
                 1825_C1U_as 
               
               
                   
               
               
                 AD-206219.1 
                 A-407982.1 
                 UGCUGAAGACAUUCAG 
                 109 
                 NM__008878.2_1967- 
                 A-407983.1 
                 UGGACCUGAAUGUCUUCAGCAAG 
                 199 
                 NM__008878.2_1965- 
               
               
                   
                   
                 GUCCA 
                   
                 1987_C21A_s 
                   
                   
                   
                 1987_G1U_as 
               
               
                   
               
               
                 AD-205227.1 
                 A-406000.1 
                 CCUCUCUGAGCUGCCG 
                 110 
                 NM__008878.2_706- 
                 A-406001.1 
                 UUAUCCGGCAGCUCAGAGAGGAA 
                 200 
                 NM__008878.2_704- 
               
               
                   
                   
                 GAUAA 
                   
                 726_G21A_s 
                   
                   
                   
                 726_C1U_as 
               
               
                   
               
               
                 AD-204943.1 
                 A-405440.1 
                 UAGUGUGGCCCUAGCA 
                 111 
                 NM__008878.2_382- 
                 A-405441.1 
                 UAGAGUGCUAGGGCCACACUAAG 
                 201 
                 NM__008878.2_380- 
               
               
                   
                   
                 CUCUA 
                   
                 402_C21A_s 
                   
                   
                   
                 402_G1U_as 
               
               
                   
               
               
                 AD-204882.1 
                 A-405318.1 
                 GGCCAUGAUGGCUUUU 
                 112 
                 NM__008878.2_301- 
                 A-405319.1 
                 UUAGUAAAAGCCAUCAUGGCCUG 
                 202 
                 NM__008878.2_299- 
               
               
                   
                   
                 ACUAA 
                   
                 321_C21A_s 
                   
                   
                   
                 321_G1U_as 
               
               
                   
               
               
                 AD-205214.1 
                 A-405974.1 
                 AGAUUGAGGAUUUCCU 
                 113 
                 NM__008878.2_693- 
                 A-405975.1 
                 UAGAGAGGAAAUCCUCAAUCUUC 
                 203 
                 NM__008878.2_691- 
               
               
                   
                   
                 CUCUA 
                   
                 713_G21A_s 
                   
                   
                   
                 713_C1U_as 
               
               
                   
               
               
                 AD-206056.1 
                 A-407656.1 
                 GGGUCCCUCCACAUUU 
                 114 
                 NM__008878.2_1802 
                 A-407657.1 
                 UUUACAAAUGUGGAGGGACCCUA 
                 204 
                 NM__008878.2_1800- 
               
               
                   
                   
                 GUAAA 
                   
                 -1822_G21A_s 
                   
                   
                   
                 1822_C1U_as 
               
               
                   
               
               
                 AD-205456.1 
                 A-406458.1 
                 AGUGGAACGUGUCCGA 
                 115 
                 NM__008878.2_969- 
                 A-406459.1 
                 UUACCUCGGACACGUUCCACUCA 
                 205 
                 NM__008878.2_967- 
               
               
                   
                   
                 GGUAA 
                   
                 989_C21A_s 
                   
                   
                   
                 989_G1U_as 
               
               
                   
               
               
                 AD-205464.1 
                 A-406474.1 
                 GUGUCCGAGGUACUAG 
                 116 
                 NM__008878.2_977- 
                 A-406475.1 
                 UUUGGCUAGUACCUCGGACACGU 
                 206 
                 NM__008878.2_975- 
               
               
                   
                   
                 CCAAA 
                   
                 997_C21A_s 
                   
                   
                   
                 997_G1U_as 
               
               
                   
               
               
                 AD-205401.1 
                 A-406348.1 
                 UGCUGGAGCAACCUGA 
                 117 
                 NM__008878.2_885- 
                 A-406349.1 
                 UUAUCUCAGGUUGCUCCAGCAGG 
                 207 
                 NM__008878.2_883- 
               
               
                   
                   
                 GAUAA 
                   
                 905_C21A_s 
                   
                   
                   
                 905_G1U_as 
               
               
                   
               
               
                 AD-205034.1 
                 A-405620.1 
                 CCCGCAUCUACUGAGC 
                 118 
                 NM__008878.2_481- 
                 A-405621.1 
                 AAAUGGCUCAGUAGAUGCGGGAG 
                 208 
                 NM__008878.2_479- 
               
               
                   
                   
                 CAUUU 
                   
                 501_s 
                   
                   
                   
                 501_as 
               
               
                   
               
            
           
         
       
     
     
       
         
           
               
             
               
                 TABLE 3 
               
             
            
               
                   
               
               
                 Modified Sense and Antisense Strand Sequences of SERPINF2 dsRNA Agents 
               
            
           
           
               
               
               
               
               
               
               
               
               
               
            
               
                   
                 Sense  
                   
                 SEQ 
                   
                   
                 SEQ 
                   
                 SEQ 
                   
               
               
                   
                 Oligo 
                 Modified Sense Sequence 
                 ID 
                 Antisense 
                 Modified Antisense Sequence 
                 ID 
                 mRNA target sequence 
                 ID 
                   
               
               
                 Duplex Name 
                 Name 
                 5′ to 3′  
                 NO: 
                 Oligo Name 
                 5′ to 3′  
                 NO: 
                 5′ to 3′  
                 NO: 
                 Range 
               
               
                   
               
               
                 AD-203163.1 
                 A-401843.1 
                 usgsuaagGfuUfUfUfuguagugauu 
                 209 
                 A-401844.1 
                 asAfsucaCfuAfCfaaaaAfcCfuuacasa 
                 299 
                 UUUGUAAGGUUUUUGU 
                 389 
                 2231-2253 
               
               
                   
                   
                 L96 
                   
                   
                 sa 
                   
                 AGUGAUU 
                   
                   
               
               
                   
               
               
                 AD-203147.1 
                 A-401811.1 
                 gscsuccuUfaAfGfGfcucuuuugua 
                 210 
                 A-401812.1 
                 usAfscaaAfaGfAfgccuUfaAfggagcsu 
                 300 
                 GAGCUCCUUAAGGCUC 
                 390 
                 2214-2236 
               
               
                   
                   
                 L96 
                   
                   
                 sc 
                   
                 UUUUGUA 
                   
                   
               
               
                   
               
               
                 AD-202129.1 
                 A-399775.1 
                 ascscuggCfaAfAfCfaucaaccaauL 
                 211 
                 A-399776.1 
                 asUfsuggUfuGfAfuguuUfgCfcaggusc 
                 301 
                 UGACCUGGCAAACAUC 
                 391 
                 674-696 
               
               
                   
                   
                 96 
                   
                   
                 sa 
                   
                 AACCAAU 
                   
                   
               
               
                   
               
               
                 AD-203171.1 
                 A-401859.1 
                 usasgugaUfuUfUfUfaugccaccua 
                 212 
                 A-401860.1 
                 usAfsgguGfgCfAfuaaaAfaUfcacuasc 
                 302 
                 UGUAGUGAUUUUUAUG 
                 392 
                 2244-2266 
               
               
                   
                   
                 L96 
                   
                   
                 sa 
                   
                 CCACCUG 
                   
                   
               
               
                   
               
               
                 AD-202239.1 
                 A-399995.1 
                 asascaagUfuUfGfAfcccgagccuu 
                 213 
                 A-399996.1 
                 asAfsggcUfcGfGfgucaAfaCfuuguusc 
                 303 
                 GGAACAAGUUUGACCC 
                 393 
                 802-824 
               
               
                   
                   
                 L96 
                   
                   
                 Sc 
                   
                 GAGCCUU 
                   
                   
               
               
                   
               
               
                 AD-202137.1 
                 A-399791.1 
                 asascaucAfaCfCfAfaugggugaaa 
                 214 
                 A-399792.1 
                 usUfsucaCfcCfAfuuggUfuGfauguusu 
                 304 
                 CAAACAUCAACCAAUG 
                 394 
                 682-704 
               
               
                   
                   
                 L96 
                   
                   
                 sg 
                   
                 GGUGAAG 
                   
                   
               
               
                   
               
               
                 AD-202196.1 
                 A-399909.1 
                 gsusugcuUfcUfCfCfucaacgccau 
                 215 
                 A-399910.1 
                 asUfsggcGfuUfGfaggaGfaAfgcaacsa 
                 305 
                 GUGUUGCUUCUCCUCA 
                 395 
                 759-781 
               
               
                   
                   
                 L96 
                   
                   
                 Sc 
                   
                 ACGCCAU 
                   
                   
               
               
                   
               
               
                 AD-203162.1 
                 A-401841.1 
                 ususguaaGfgUfUfUfuuguagugau 
                 216 
                 A-401842.1 
                 asUfscacUfaCfAfaaaaCfcUfuacaasa 
                 306 
                 UUUUGUAAGGUUUUUG 
                 396 
                 2230-2252 
               
               
                   
                   
                 L96 
                   
                   
                 sa 
                   
                 UAGUGAU 
                   
                   
               
               
                   
               
               
                 AD-202130.1 
                 A-399777.1 
                 cscsuggcAfaAfCfAfucaaccaauaL 
                 217 
                 A-399778.1 
                 usAfsuugGfuUfGfauguUfuGfccaggsu 
                 307 
                 GACCUGGCAAACAUCA 
                 397 
                 675-697 
               
               
                   
                   
                 96 
                   
                   
                 Sc 
                   
                 ACCAAUG 
                   
                   
               
               
                   
               
               
                 AD-203169.1 
                 A-401855.1 
                 usgsuaguGfaUfUfUfuuaugccaca 
                 218 
                 A-401856.1 
                 usGfsuggCfaUfAfaaaaUfcAfcuacasa 
                 308 
                 UUUGUAGUGAUUUUUA 
                 398 
                 2242-2264 
               
               
                   
                   
                 L96 
                   
                   
                 sa 
                   
                 UGCCACC 
                   
                   
               
               
                   
               
               
                 AD-202052.1 
                 A-399621.1 
                 usgscagaAfaGfGfAfuuucccauca 
                 219 
                 A-399622.1 
                 usGfsaugGfgAfAfauccUfuUfcugcasg 
                 309 
                 CCUGCAGAAAGGAUUU 
                 399 
                 578-600 
               
               
                   
                   
                 L96 
                   
                   
                 sg 
                   
                 CCCAUCA 
                   
                   
               
               
                   
               
               
                 AD-203184.1 
                 A-401885.1 
                 gscscaccUfgAfAfUfaaagaaugaa 
                 220 
                 A-401886.1 
                 usUfscauUfcUfUfuauuCfaGfguggcsa 
                 310 
                 AUGCCACCUGAAUAAA 
                 400 
                 2257-2279 
               
               
                   
                   
                 L96 
                   
                   
                 su 
                   
                 GAAUGAA 
                   
                   
               
               
                   
               
               
                 AD-202492.1 
                 A-400501.1 
                 cscsuaagCfuGfUfAfucugaaacaa 
                 221 
                 A-400502.1 
                 usUfsguuUfcAfGfauacAfgCfuuaggsc 
                 311 
                 UGCCUAAGCUGUAUCU 
                 401 
                 1084-1106 
               
               
                   
                   
                 L96 
                   
                   
                 sa 
                   
                 GAAACAC 
                   
                   
               
               
                   
               
               
                 AD-202491.1 
                 A-400499.1 
                 gscscuaaGfcUfGfUfaucugaaaca 
                 222 
                 A-400500.1 
                 usGfsuuuCfaGfAfuacaGfcUfuaggcsa 
                 312 
                 CUGCCUAAGCUGUAUC 
                 402 
                 1083-1105 
               
               
                   
                   
                 L96 
                   
                   
                 sg 
                   
                 UGAAACA 
                   
                   
               
               
                   
               
               
                 AD-202050.1 
                 A-399617.1 
                 cscsugcaGfaAfAfGfgauuucccau 
                 223 
                 A-399618.1 
                 asUfsgggAfaAfUfccuuUfcUfgcaggsu 
                 313 
                 UACCUGCAGAAAGGAU 
                 403 
                 576-598 
               
               
                   
                   
                 L96 
                   
                   
                 sa 
                   
                 UUCCCAU 
                   
                   
               
               
                   
               
               
                 AD-202197.1 
                 A-399911.1 
                 ususgcuuCfuCfCfUfcaacgccaua 
                 224 
                 A-399912.1 
                 usAfsuggCfgUfUfgaggAfgAfagcaasc 
                 314 
                 UGUUGCUUCUCCUCAA 
                 404 
                 760-782 
               
               
                   
                   
                 L96 
                   
                   
                 sa 
                   
                 CGCCAUC 
                   
                   
               
               
                   
               
               
                 AD-202192.1 
                 A-399901.1 
                 cscsguguUfgCfUfUfcuccucaaca 
                 225 
                 A-399902.1 
                 usGfsuugAfgGfAfgaagCfaAfcacggsu 
                 315 
                 CACCGUGUUGCUUCUC 
                 405 
                 755-777 
               
               
                   
                   
                 L96 
                   
                   
                 sg 
                   
                 CUCAACG 
                   
                   
               
               
                   
               
               
                 AD-203148.1 
                 A-401813.1 
                 csusccuuAfaGfGfCfucuuuuguaa 
                 226 
                 A-401814.1 
                 usUfsacaAfaAfGfagccUfuAfaggagsc 
                 316 
                 AGCUCCUUAAGGCUCU 
                 406 
                 2215-2237 
               
               
                   
                   
                 L96 
                   
                   
                 su 
                   
                 UUUGUAA 
                   
                   
               
               
                   
               
               
                 AD-202499.1 
                 A-400515.1 
                 usgsuaucUfgAfAfAfcaccaaauga 
                 227 
                 A-400516.1 
                 usCfsauuUfgGfUfguuuCfaGfauacasg 
                 317 
                 GCUGUAUCUGAAACAC 
                 407 
                 1091-1113 
               
               
                   
                   
                 L96 
                   
                   
                 Sc 
                   
                 CAAAUGG 
                   
                   
               
               
                   
               
               
                 AD-202054.1 
                 A-399625.1 
                 csasgaaaGfgAfUfUfucccaucaaaL 
                 228 
                 A-399626.1 
                 usUfsugaUfgGfGfaaauCfcUfuucugsc 
                 318 
                 UGCAGAAAGGAUUUCC 
                 408 
                 580-602 
               
               
                   
                   
                 96 
                   
                   
                 sa 
                   
                 CAUCAAA 
                   
                   
               
               
                   
               
               
                 AD-202372.1 
                 A-400261.1 
                 gsasgcuuUfgUfGfGfuccuuguaca 
                 229 
                 A-400262.1 
                 usGfsuacAfaGfGfaccaCfaAfagcucsas 
                 319 
                 AUGAGCUUUGUGGUCC 
                 409 
                 963-985 
               
               
                   
                   
                 L96 
                   
                   
                 u 
                   
                 UUGUACC 
                   
                   
               
               
                   
               
               
                 AD-202796.1 
                 A-401109.1 
                 csuscaugUfgAfCfUfcuuuccaaca 
                 230 
                 A-401110.1 
                 usGfsuugGfaAfAfgaguCfaCfaugagsu 
                 320 
                 CACUCAUGUGACUCUU 
                 410 
                 1601-1623 
               
               
                   
                   
                 L96 
                   
                   
                 sg 
                   
                 UCCAACC 
                   
                   
               
               
                   
               
               
                 AD-206288.1 
                 A-408120.1 
                 gscsucccAfaGfAfCfucuuuuguaa 
                 231 
                 A-408121.1 
                 usUfsacaAfaAfGfagucUfuGfggagcsu 
                 321 
                 UAGCUCCCAAGACUCU 
                 411 
                 2123-2145 
               
               
                   
                   
                 L96 
                   
                   
                 sa 
                   
                 UUUGUAA 
                   
                   
               
               
                   
               
               
                 AD-202191.1 
                 A-399899.1 
                 ascscgugUfuGfCfUfucuccucaaa 
                 232 
                 A-399900.1 
                 usUfsugaGfgAfGfaagcAfaCfacggusg 
                 322 
                 ACACCGUGUUGCUUCU 
                 412 
                 754-776 
               
               
                   
                   
                 L96 
                   
                   
                 su 
                   
                 CCUCAAC 
                   
                   
               
               
                   
               
               
                 AD-203177.1 
                 A-401871.1 
                 ususuuuaUfgCfCfAfccugaauaaa 
                 233 
                 A-401872.1 
                 usUfsuauUfcAfGfguggCfaUfaaaaasu 
                 323 
                 GAUUUUUAUGCCACCU 
                 413 
                 2250-2272 
               
               
                   
                   
                 L96 
                   
                   
                 sc 
                   
                 GAAUAAA 
                   
                   
               
               
                   
               
               
                 AD-202489.1 
                 A-400495.1 
                 csusgccuAfaGfCfUfguaucugaaa 
                 234 
                 A-400496.1 
                 usUfsucaGfaUfAfcagcUfuAfggcagsc 
                 324 
                 GGCUGCCUAAGCUGUA 
                 414 
                 1081-1103 
               
               
                   
                   
                 L96 
                   
                   
                 sc 
                   
                 UCUGAAA 
                   
                   
               
               
                   
               
               
                 AD-202387.1 
                 A-400291.1 
                 usgsuaccCfaCfCfCfacuuugaaua 
                 235 
                 A-400292.1 
                 usAfsuucAfaAfGfugggUfgGfguacasa 
                 325 
                 CUUGUACCCACCCACU 
                 415 
                  978-1000 
               
               
                   
                   
                 L96 
                   
                   
                 sg 
                   
                 UUGAAUG 
                   
                   
               
               
                   
               
               
                 AD-202399.1 
                 A-400315.1 
                 csusuugaAfuGfGfAfacguguccca 
                 236 
                 A-400316.1 
                 usGfsggaCfaCfGfuuccAfuUfcaaagsu 
                 326 
                 CACUUUGAAUGGAACG 
                 416 
                  990-1012 
               
               
                   
                   
                 L96 
                   
                   
                 sg 
                   
                 UGUCCCA 
                   
                   
               
               
                   
               
               
                 AD-202386.1 
                 A-400289.1 
                 ususguacCfcAfCfCfcacuuugaau 
                 237 
                 A-400290.1 
                 asUfsucaAfaGfUfggguGfgGfuacaasg 
                 327 
                 CCUUGUACCCACCCACU 
                 417 
                 977-999 
               
               
                   
                   
                 L96 
                   
                   
                 sg 
                   
                 UUGAAU 
                   
                   
               
               
                   
               
               
                 AD-202053.1 
                 A-399623.1 
                 gscsagaaAfgGfAfUfuucccaucaa 
                 238 
                 A-399624.1 
                 usUfsgauGfgGfAfaaucCfuUfucugcsa 
                 328 
                 CUGCAGAAAGGAUUUC 
                 418 
                 579-601 
               
               
                   
                   
                 L96 
                   
                   
                 sg 
                   
                 CCAUCAA 
                   
                   
               
               
                   
               
               
                 AD-203166.1 
                 A-401849.1 
                 ususuuguAfgUfGfAfuuuuuaugca 
                 239 
                 A-401850.1 
                 usGfscauAfaAfAfaucaCfuAfcaaaasa 
                 329 
                 GUUUUUGUAGUGAUUU 
                 419 
                 2239-2261 
               
               
                   
                   
                 L96 
                   
                   
                 sc 
                   
                 UUAUGCC 
                   
                   
               
               
                   
               
               
                 AD-202827.1 
                 A-401171.1 
                 gsasggccAfuUfCfUfuucccaacaa 
                 240 
                 A-401172.1 
                 usUfsguuGfgGfAfaagaAfuGfgccucsu 
                 330 
                 GAGAGGCCAUUCUUUC 
                 420 
                 1663-1685 
               
               
                   
                   
                 L96 
                   
                   
                 sc 
                   
                 CCAACAC 
                   
                   
               
               
                   
               
               
                 AD-205123.1 
                 A-405796.1 
                 gsasuuucCfuGfGfAfgcaaucggaa 
                 241 
                 A-405797.1 
                 usUfsccgAfuUfGfcuccAfgGfaaaucsg 
                 331 
                 ACGAUUUCCUGGAGCA 
                 421 
                 576-598 
               
               
                   
                   
                 L96 
                   
                   
                 su 
                   
                 AUCGGAA 
                   
                   
               
               
                   
               
               
                 AD-202490.1 
                 A-400497.1 
                 usgsccuaAfgCfUfGfuaucugaaac 
                 242 
                 A-400498.1 
                 gsUfsuucAfgAfUfacagCfuUfaggcasg 
                 332 
                 GCUGCCUAAGCUGUAU 
                 422 
                 1082-1104 
               
               
                   
                   
                 L96 
                   
                   
                 Sc 
                   
                 CUGAAAC 
                   
                   
               
               
                   
               
               
                 AD-202051.1 
                 A-399619.1 
                 csusgcagAfaAfGfGfauuucccaua 
                 243 
                 A-399620.1 
                 usAfsuggGfaAfAfuccuUfuCfugcagsg 
                 333 
                 ACCUGCAGAAAGGAUU 
                 423 
                 577-599 
               
               
                   
                   
                 L96 
                   
                   
                 su 
                   
                 UCCCAUC 
                   
                   
               
               
                   
               
               
                 AD-202894.1 
                 A-401305.1 
                 asgsggagGfuGfCfUfucuaguucua 
                 244 
                 A-401306.1 
                 usAfsgaaCfuAfGfaagcAfcCfucccusc 
                 334 
                 GGAGGGAGGUGCUUCU 
                 424 
                 1813-1835 
               
               
                   
                   
                 L96 
                   
                   
                 sc 
                   
                 AGUUCUG 
                   
                   
               
               
                   
               
               
                 AD-202494.1 
                 A-400505.1 
                 usasagcuGfuAfUfCfugaaacacca 
                 245 
                 A-400506.1 
                 usGfsgugUfuUfCfagauAfcAfgcuuasg 
                 335 
                 CCUAAGCUGUAUCUGA 
                 425 
                 1086-1108 
               
               
                   
                   
                 L96 
                   
                   
                 sg 
                   
                 AACACCA 
                   
                   
               
               
                   
               
               
                 AD-202905.1 
                 A-401327.1 
                 uscsuaguUfcUfGfCfcaggagacaa 
                 246 
                 A-401328.1 
                 usUfsgucUfcCfUfggcaGfaAfcuagasa 
                 336 
                 CUUCUAGUUCUGCCAG 
                 426 
                 1824-1846 
               
               
                   
                   
                 L96 
                   
                   
                 sg 
                   
                 GAGACAG 
                   
                   
               
               
                   
               
               
                 AD-202904.1 
                 A-401325.1 
                 ususcuagUfuCfUfGfccaggagaca 
                 247 
                 A-401326.1 
                 usGfsucuCfcUfGfgcagAfaCfuagaasg 
                 337 
                 GCUUCUAGUUCUGCCA 
                 427 
                 1823-1845 
               
               
                   
                   
                 L96 
                   
                   
                 Sc 
                   
                 GGAGACA 
                   
                   
               
               
                   
               
               
                 AD-203151.1 
                 A-401819.1 
                 csusuaagGfcUfCfUfuuuguaaggu 
                 248 
                 A-401820.1 
                 asCfscuuAfcAfAfaagaGfcCfuuaagsg 
                 338 
                 UCCUUAAGGCUCUUUU 
                 428 
                 2218-2240 
               
               
                   
                   
                 L96 
                   
                   
                 sa 
                   
                 GUAAGGU 
                   
                   
               
               
                   
               
               
                 AD-202406.1 
                 A-400329.1 
                 usgsgaacGfuGfUfCfccagguacua 
                 249 
                 A-400330.1 
                 usAfsguaCfcUfGfggacAfcGfuuccasu 
                 339 
                 AAUGGAACGUGUCCCA 
                 429 
                  997-1019 
               
               
                   
                   
                 L96 
                   
                   
                 su 
                   
                 GGUACUG 
                   
                   
               
               
                   
               
               
                 AD-202913.1 
                 A-401343.1 
                 usgsccagGfaGfAfCfagguuagcua 
                 250 
                 A-401344.1 
                 usAfsgcuAfaCfCfugucUfcCfuggcasg 
                 340 
                 UCUGCCAGGAGACAGG 
                 430 
                 1832-1854 
               
               
                   
                   
                 L96 
                   
                   
                 sa 
                   
                 UUAGCUG 
                   
                   
               
               
                   
               
               
                 AD-202277.1 
                 A-400071.1 
                 cscsaccuGfgAfCfGfagcaguucaa 
                 251 
                 A-400072.1 
                 usUfsgaaCfuGfCfucguCfcAfgguggsa 
                 341 
                 UUCCACCUGGACGAGC 
                 431 
                 840-862 
               
               
                   
                   
                 L96 
                   
                   
                 sa 
                   
                 AGUUCAC 
                   
                   
               
               
                   
               
               
                 AD-202133.1 
                 A-399783.1 
                 gsgscaaaCfaUfCfAfaccaaugggu 
                 252 
                 A-399784.1 
                 asCfsccaUfuGfGfuugaUfgUfuugccsa 
                 342 
                 CUGGCAAACAUCAACC 
                 432 
                 678-700 
               
               
                   
                   
                 L96 
                   
                   
                 sg 
                   
                 AAUGGGU 
                   
                   
               
               
                   
               
               
                 AD-202895.1 
                 A-401307.1 
                 gsgsgaggUfgCfUfUfcuaguucuga 
                 253 
                 A-401308.1 
                 usCfsagaAfcUfAfgaagCfaCfcucccsu 
                 343 
                 GAGGGAGGUGCUUCUA 
                 433 
                 1814-1836 
               
               
                   
                   
                 L96 
                   
                   
                 sc 
                   
                 GUUCUGC 
                   
                   
               
               
                   
               
               
                 AD-202398.1 
                 A-400313.1 
                 ascsuuugAfaUfGfGfaacgugucca 
                 254 
                 A-400314.1 
                 usGfsgacAfcGfUfuccaUfuCfaaagusg 
                 344 
                 CCACUUUGAAUGGAAC 
                 434 
                  989-1011 
               
               
                   
                   
                 L96 
                   
                   
                 sg 
                   
                 GUGUCCC 
                   
                   
               
               
                   
               
               
                 AD-202136.1 
                 A-399789.1 
                 asasacauCfaAfCfCfaaugggugaaL 
                 255 
                 A-399790.1 
                 usUfscacCfcAfUfugguUfgAfuguuusg 
                 345 
                 GCAAACAUCAACCAAU 
                 435 
                 681-703 
               
               
                   
                   
                 96 
                   
                   
                 sc 
                   
                 GGGUGAA 
                   
                   
               
               
                   
               
               
                 AD-202131.1 
                 A-399779.1 
                 csusggcaAfaCfAfUfcaaccaaugaL 
                 256 
                 A-399780.1 
                 usCfsauuGfgUfUfgaugUfuUfgccagsg 
                 346 
                 ACCUGGCAAACAUCAA 
                 436 
                 676-698 
               
               
                   
                   
                 96 
                   
                   
                 su 
                   
                 CCAAUGG 
                   
                   
               
               
                   
               
               
                 AD-202393.1 
                 A-400303.1 
                 csascccaCfuUfUfGfaauggaacgu 
                 257 
                 A-400304.1 
                 asCfsguuCfcAfUfucaaAfgUfgggugsg 
                 347 
                 CCCACCCACUUUGAAU 
                 437 
                  984-1006 
               
               
                   
                   
                 L96 
                   
                   
                 sg 
                   
                 GGAACGU 
                   
                   
               
               
                   
               
               
                 AD-201919.1 
                 A-399355.1 
                 csuscccuGfgUfGfGfcucaaacgua 
                 258 
                 A-399356.1 
                 usAfscguUfuGfAfgccaCfcAfgggagsa 
                 348 
                 UUCUCCCUGGUGGCUC 
                 438 
                 357-379 
               
               
                   
                   
                 L96 
                   
                   
                 sa 
                   
                 AAACGUC 
                   
                   
               
               
                   
               
               
                 AD-202787.1 
                 A-401091.1 
                 gscscucuCfcAfCfUfcaugugacua 
                 259 
                 A-401092.1 
                 usAfsgucAfcAfUfgaguGfgAfgaggcsu 
                 349 
                 CAGCCUCUCCACUCAU 
                 439 
                 1592-1614 
               
               
                   
                   
                 L96 
                   
                   
                 sg 
                   
                 GUGACUC 
                   
                   
               
               
                   
               
               
                 AD-205414.1 
                 A-406374.1 
                 usgsagauAfcAfGfGfuggcucauuu 
                 260 
                 A-406375.1 
                 asAfsaugAfgCfCfaccuGfuAfucucasg 
                 350 
                 CCUGAGAUACAGGUGG 
                 440 
                 896-918 
               
               
                   
                   
                 L96 
                   
                   
                 sg 
                   
                 CUCAUUU 
                   
                   
               
               
                   
               
               
                 AD-205279.1 
                 A-406104.1 
                 csgsguuuCfuGfGfAfggaccaaguu 
                 261 
                 A-406105.1 
                 asAfscuuGfgUfCfcuccAfgAfaaccgsu 
                 351 
                 CACGGUUUCUGGAGGA 
                 441 
                 761-783 
               
               
                   
                   
                 L96 
                   
                   
                 sg 
                   
                 CCAAGUU 
                   
                   
               
               
                   
               
               
                 AD-202373.1 
                 A-400263.1 
                 asgscuuuGfuGfGfUfccuuguacca 
                 262 
                 A-400264.1 
                 usGfsguaCfaAfGfgaccAfcAfaagcusc 
                 352 
                 UGAGCUUUGUGGUCCU 
                 442 
                 964-986 
               
               
                   
                   
                 L96 
                   
                   
                 sa 
                   
                 UGUACCC 
                   
                   
               
               
                   
               
               
                 AD-202198.1 
                 A-399913.1 
                 usgscuucUfcCfUfCfaacgccaucaL 
                 263 
                 A-399914.1 
                 usGfsaugGfcGfUfugagGfaGfaagcasa 
                 353 
                 GUUGCUUCUCCUCAAC 
                 443 
                 761-783 
               
               
                   
                   
                 96 
                   
                   
                 sc 
                   
                 GCCAUCC 
                   
                   
               
               
                   
               
               
                 AD-202420.1 
                 A-400357.1 
                 gsgsuacuGfgCfCfAfaccugaguua 
                 264 
                 A-400358.1 
                 usAfsacuCfaGfGfuuggCfcAfguaccsu 
                 354 
                 CAGGUACUGGCCAACC 
                 444 
                 1011-1033 
               
               
                   
                   
                 L96 
                   
                   
                 sg 
                   
                 UGAGUUG 
                   
                   
               
               
                   
               
               
                 AD-202199.1 
                 A-399915.1 
                 gscsuucuCfcUfCfAfacgccauccaL 
                 265 
                 A-399916.1 
                 usGfsgauGfgCfGfuugaGfgAfgaagcsa 
                 355 
                 UUGCUUCUCCUCAACG 
                 445 
                 762-784 
               
               
                   
                   
                 96 
                   
                   
                 sa 
                   
                 CCAUCCA 
                   
                   
               
               
                   
               
               
                 AD-204885.1 
                 A-405324.1 
                 csasugauGfgCfUfUfuuacuacuga 
                 266 
                 A-405325.1 
                 usCfsaguAfgUfAfaaagCfcAfucaugsg 
                 356 
                 GCCAUGAUGGCUUUUA 
                 446 
                 302-324 
               
               
                   
                   
                 L96 
                   
                   
                 sc 
                   
                 CUACUGA 
                   
                   
               
               
                   
               
               
                 AD-205284.1 
                 A-406114.1 
                 uscsuggaGfgAfCfCfaaguuugaca 
                 267 
                 A-406115.1 
                 usGfsucaAfaCfUfugguCfcUfccagasa 
                 357 
                 UUUCUGGAGGACCAAG 
                 447 
                 766-788 
               
               
                   
                   
                 L96 
                   
                   
                 sa 
                   
                 UUUGACC 
                   
                   
               
               
                   
               
               
                 AD-205458.1 
                 A-406462.1 
                 usgsgaacGfuGfUfCfcgagguacua 
                 268 
                 A-406463.1 
                 usAfsguaCfcUfCfggacAfcGfuuccasc 
                 358 
                 AGUGGAACGUGUCCGA 
                 448 
                 969-991 
               
               
                   
                   
                 L96 
                   
                   
                 su 
                   
                 GGUACUA 
                   
                   
               
               
                   
               
               
                 AD-205413.1 
                 A-406372.1 
                 csusgagaUfaCfAfGfguggcucauu 
                 269 
                 A-406373.1 
                 asAfsugaGfcCfAfccugUfaUfcucagsg 
                 359 
                 ACCUGAGAUACAGGUG 
                 449 
                 895-917 
               
               
                   
                   
                 L96 
                   
                   
                 su 
                   
                 GCUCAUU 
                   
                   
               
               
                   
               
               
                 AD-206058.1 
                 A-407660.1 
                 gsuscccuCfcAfCfAfuuuguaagca 
                 270 
                 A-407661.1 
                 usGfscuuAfcAfAfauguGfgAfgggacsc 
                 360 
                 GGGUCCCUCCACAUUU 
                 450 
                 1802-1824 
               
               
                   
                   
                 L96 
                   
                   
                 sc 
                   
                 GUAAGCA 
                   
                   
               
               
                   
               
               
                 AD-205411.1 
                 A-406368.1 
                 ascscugaGfaUfAfCfagguggcuca 
                 271 
                 A-406369.1 
                 usGfsagcCfaCfCfuguaUfcUfcaggusu 
                 361 
                 CAACCUGAGAUACAGG 
                 451 
                 893-915 
               
               
                   
                   
                 L96 
                   
                   
                 sg 
                   
                 UGGCUCA 
                   
                   
               
               
                   
               
               
                 AD-202407.1 
                 A-400331.1 
                 gsgsaacgUfgUfCfCfcagguacuga 
                 272 
                 A-400332.1 
                 usCfsaguAfcCfUfgggaCfaCfguuccsa 
                 362 
                 AUGGAACGUGUCCCAG 
                 452 
                  998-1020 
               
               
                   
                   
                 L96 
                   
                   
                 su 
                   
                 GUACUGG 
                   
                   
               
               
                   
               
               
                 AD-205459.1 
                 A-406464.1 
                 gsgsaacgUfgUfCfCfgagguacuaa 
                 273 
                 A-406465.1 
                 usUfsaguAfcCfUfcggaCfaCfguuccsa 
                 363 
                 GUGGAACGUGUCCGAG 
                 453 
                 970-992 
               
               
                   
                   
                 L96 
                   
                   
                 sc 
                   
                 GUACUAG 
                   
                   
               
               
                   
               
               
                 AD-205455.1 
                 A-406456.1 
                 gsasguggAfaCfGfUfguccgaggua 
                 274 
                 A-406457.1 
                 usAfsccuCfgGfAfcacgUfuCfcacucsa 
                 364 
                 UUGAGUGGAACGUGUC 
                 454 
                 966-988 
               
               
                   
                   
                 L96 
                   
                   
                 sa 
                   
                 CGAGGUA 
                   
                   
               
               
                   
               
               
                 AD-205264.1 
                 A-406074.1 
                 csgsccauCfcAfCfUfuucacgguuu 
                 275 
                 A-406075.1 
                 asAfsaccGfuGfAfaaguGfgAfuggcgsu 
                 365 
                 AACGCCAUCCACUUUC 
                 455 
                 746-768 
               
               
                   
                   
                 L96 
                   
                   
                 su 
                   
                 ACGGUUU 
                   
                   
               
               
                   
               
               
                 AD-204884.1 
                 A-405322.1 
                 cscsaugaUfgGfCfUfuuuacuacua 
                 276 
                 A-405323.1 
                 usAfsguaGfuAfAfaagcCfaUfcauggsc 
                 366 
                 GGCCAUGAUGGCUUUU 
                 456 
                 301-323 
               
               
                   
                   
                 L96 
                   
                   
                 sc 
                   
                 ACUACUG 
                   
                   
               
               
                   
               
               
                 AD-204888.1 
                 A-405330.1 
                 gsasuggcUfuUfUfAfcuacugaccu 
                 277 
                 A-405331.1 
                 asGfsgucAfgUfAfguaaAfaGfccaucsa 
                 367 
                 AUGAUGGCUUUUACUA 
                 457 
                 305-327 
               
               
                   
                   
                 L96 
                   
                   
                 su 
                   
                 CUGACCU 
                   
                   
               
               
                   
               
               
                 AD-205270.1 
                 A-406086.1 
                 cscsacuuUfcAfCfGfguuucuggaa 
                 278 
                 A-406087.1 
                 usUfsccaGfaAfAfccguGfaAfaguggsa 
                 368 
                 AUCCACUUUCACGGUU 
                 458 
                 752-774 
               
               
                   
                   
                 L96 
                   
                   
                 su 
                   
                 UCUGGAG 
                   
                   
               
               
                   
               
               
                 AD-204894.1 
                 A-405342.1 
                 ususuuacUfaCfUfGfaccuguucua 
                 279 
                 A-405343.1 
                 usAfsgaaCfaGfGfucagUfaGfuaaaasg 
                 369 
                 GCUUUUACUACUGACC 
                 459 
                 311-333 
               
               
                   
                   
                 L96 
                   
                   
                 sc 
                   
                 UGUUCUC 
                   
                   
               
               
                   
               
               
                 AD-205273.1 
                 A-406092.1 
                 csusuucaCfgGfUfUfucuggaggaa 
                 280 
                 A-406093.1 
                 usUfsccuCfcAfGfaaacCfgUfgaaagsu 
                 370 
                 CACUUUCACGGUUUCU 
                 460 
                 755-777 
               
               
                   
                   
                 L96 
                   
                   
                 sg 
                   
                 GGAGGAC 
                   
                   
               
               
                   
               
               
                 AD-206218.1 
                 A-407980.1 
                 ususgcugAfaGfAfCfauucagguca 
                 281 
                 A-407981.1 
                 usGfsaccUfgAfAfugucUfuCfagcaasg 
                 371 
                 CCUUGCUGAAGACAUU 
                 461 
                 1964-1986 
               
               
                   
                   
                 L96 
                   
                   
                 sg 
                   
                 CAGGUCC 
                   
                   
               
               
                   
               
               
                 AD-202132.1 
                 A-399781.1 
                 usgsgcaaAfcAfUfCfaaccaaugga 
                 282 
                 A-399782.1 
                 usCfscauUfgGfUfugauGfuUfugccasg 
                 372 
                 CCUGGCAAACAUCAAC 
                 462 
                 677-699 
               
               
                   
                   
                 L96 
                   
                   
                 sg 
                   
                 CAAUGGG 
                   
                   
               
               
                   
               
               
                 AD-202389.1 
                 A-400295.1 
                 usascccaCfcCfAfCfuuugaaugga 
                 283 
                 A-400296.1 
                 usCfscauUfcAfAfagugGfgUfggguasc 
                 373 
                 UGUACCCACCCACUUU 
                 463 
                  980-1002 
               
               
                   
                   
                 L96 
                   
                   
                 sa 
                   
                 GAAUGGA 
                   
                   
               
               
                   
               
               
                 AD-204944.1 
                 A-405442.1 
                 asgsugugGfcCfCfUfagcacucucu 
                 284 
                 A-405443.1 
                 asGfsagaGfuGfCfuaggGfcCfacacusa 
                 374 
                 UUAGUGUGGCCCUAGC 
                 464 
                 381-403 
               
               
                   
                   
                 L96 
                   
                   
                 sa 
                   
                 ACUCUCU 
                   
                   
               
               
                   
               
               
                 AD-205463.1 
                 A-406472.1 
                 csgsugucCfgAfGfGfuacuagccaa 
                 285 
                 A-406473.1 
                 usUfsggcUfaGfUfaccuCfgGfacacgsu 
                 375 
                 AACGUGUCCGAGGUAC 
                 465 
                 974-996 
               
               
                   
                   
                 L96 
                   
                   
                 su 
                   
                 UAGCCAA 
                   
                   
               
               
                   
               
               
                 AD-206057.1 
                 A-407658.1 
                 gsgsucccUfcCfAfCfauuuguaaga 
                 286 
                 A-407659.1 
                 usCfsuuaCfaAfAfugugGfaGfggaccsc 
                 376 
                 AGGGUCCCUCCACAUU 
                 466 
                 1801-1823 
               
               
                   
                   
                 L96 
                   
                   
                 su 
                   
                 UGUAAGC 
                   
                   
               
               
                   
               
               
                 AD-206281.1 
                 A-408106.1 
                 cscscuuuAfgCfUfCfccaagacucu 
                 287 
                 A-408107.1 
                 asGfsaguCfuUfGfggagCfuAfaagggsu 
                 377 
                 GACCCUUUAGCUCCCA 
                 467 
                 2116-2138 
               
               
                   
                   
                 L96 
                   
                   
                 sc 
                   
                 AGACUCU 
                   
                   
               
               
                   
               
               
                 AD-206059.1 
                 A-407662.1 
                 uscsccucCfaCfAfUfuuguaagcaa 
                 288 
                 A-407663.1 
                 usUfsgcuUfaCfAfaaugUfgGfagggasc 
                 378 
                 GGUCCCUCCACAUUUG 
                 468 
                 1803-1825 
               
               
                   
                   
                 L96 
                   
                   
                 sc 
                   
                 UAAGCAG 
                   
                   
               
               
                   
               
               
                 AD-206219.1 
                 A-407982.1 
                 usgscugaAfgAfCfAfuucaggucca 
                 289 
                 A-407983.1 
                 usGfsgacCfuGfAfauguCfuUfcagcasa 
                 379 
                 CUUGCUGAAGACAUUC 
                 469 
                 1965-1987 
               
               
                   
                   
                 L96 
                   
                   
                 sg 
                   
                 AGGUCCC 
                   
                   
               
               
                   
               
               
                 AD-205227.1 
                 A-406000.1 
                 cscsucucUfgAfGfCfugccggauaa 
                 290 
                 A-406001.1 
                 usUfsaucCfgGfCfagcuCfaGfagaggsa 
                 380 
                 UUCCUCUCUGAGCUGC 
                 470 
                 704-726 
               
               
                   
                   
                 L96 
                   
                   
                 sa 
                   
                 CGGAUAG 
                   
                   
               
               
                   
               
               
                 AD-204943.1 
                 A-405440.1 
                 usasguguGfgCfCfCfuagcacucua 
                 291 
                 A-405441.1 
                 usAfsgagUfgCfUfagggCfcAfcacuasa 
                 381 
                 CUUAGUGUGGCCCUAG 
                 471 
                 380-402 
               
               
                   
                   
                 L96 
                   
                   
                 sg 
                   
                 CACUCUC 
                   
                   
               
               
                   
               
               
                 AD-204882.1 
                 A-405318.1 
                 gsgsccauGfaUfGfGfcuuuuacuaa 
                 292 
                 A-405319.1 
                 usUfsaguAfaAfAfgccaUfcAfuggccsu 
                 382 
                 CAGGCCAUGAUGGCUU 
                 472 
                 299-321 
               
               
                   
                   
                 L96 
                   
                   
                 sg 
                   
                 UUACUAC 
                   
                   
               
               
                   
               
               
                 AD-205214.1 
                 A-405974.1 
                 asgsauugAfgGfAfUfuuccucucua 
                 293 
                 A-405975.1 
                 usAfsgagAfgGfAfaaucCfuCfaaucusu 
                 383 
                 GAAGAUUGAGGAUUUC 
                 473 
                 691-713 
               
               
                   
                   
                 L96 
                   
                   
                 sc 
                   
                 CUCUCUG 
                   
                   
               
               
                   
               
               
                 AD-206056.1 
                 A-407656.1 
                 gsgsguccCfuCfCfAfcauuuguaaa 
                 294 
                 A-407657.1 
                 usUfsuacAfaAfUfguggAfgGfgacccsu 
                 384 
                 UAGGGUCCCUCCACAU 
                 474 
                 1800-1822 
               
               
                   
                   
                 L96 
                   
                   
                 sa 
                   
                 UUGUAAG 
                   
                   
               
               
                   
               
               
                 AD-205456.1 
                 A-406458.1 
                 asgsuggaAfcGfUfGfuccgagguaa 
                 295 
                 A-406459.1 
                 usUfsaccUfcGfGfacacGfuUfccacusc 
                 385 
                 UGAGUGGAACGUGUCC 
                 475 
                 967-989 
               
               
                   
                   
                 L96 
                   
                   
                 sa 
                   
                 GAGGUAC 
                   
                   
               
               
                   
               
               
                 AD-205464.1 
                 A-406474.1 
                 gsusguccGfaGfGfUfacuagccaaa 
                 296 
                 A-406475.1 
                 usUfsuggCfuAfGfuaccUfcGfgacacsg 
                 386 
                 ACGUGUCCGAGGUACU 
                 476 
                 975-997 
               
               
                   
                   
                 L96 
                   
                   
                 su 
                   
                 AGCCAAC 
                   
                   
               
               
                   
               
               
                 AD-205401.1 
                 A-406348.1 
                 usgscuggAfgCfAfAfccugagauaa 
                 297 
                 A-406349.1 
                 usUfsaucUfcAfGfguugCfuCfcagcasg 
                 387 
                 CCUGCUGGAGCAACCU 
                 477 
                 883-905 
               
               
                   
                   
                 L96 
                   
                   
                 sg 
                   
                 GAGAUAC 
                   
                   
               
               
                   
               
               
                 AD-205034.1 
                 A-405620.1 
                 cscscgcaUfcUfAfCfugagccauuu 
                 298 
                 A-405621.1 
                 asAfsaugGfcUfCfaguaGfaUfgcgggsa 
                 388 
                 CUCCCGCAUCUACUGA 
                 478 
                 479-501 
               
               
                   
                   
                 L96 
                   
                   
                 sg 
                   
                 GCCAUUU 
               
               
                   
               
            
           
         
       
     
     
       
         
           
               
             
               
                 TABLE 4 
               
             
            
               
                   
               
               
                 SERPINF2 Single 10 nM and 0.1 nM Dose Screen in Hep3B cells 
               
               
                 (values indicate % of message remaining) 
               
            
           
           
               
               
               
               
               
            
               
                   
                 10 nM  
                 10 nM 
                 0.1 nM  
                 0.1 nM 
               
               
                 Duplex ID 
                 Average 
                 STDEV 
                 Average 
                 STDEV 
               
               
                   
               
            
           
           
               
               
               
               
               
            
               
                 AD-203163.1 
                 3.99 
                 1.54 
                 35.56 
                 6.61 
               
               
                 AD-203147.1 
                 4.43 
                 0.95 
                 74.45 
                 7.80 
               
               
                 AD-202129.1 
                 5.09 
                 0.65 
                 51.46 
                 11.46 
               
               
                 AD-203171.1 
                 6.47 
                 2.80 
                 70.40 
                 8.24 
               
               
                 AD-202239.1 
                 7.11 
                 3.08 
                 72.92 
                 11.49 
               
               
                 AD-202137.1 
                 8.93 
                 3.08 
                 85.09 
                 9.35 
               
               
                 AD-202196.1 
                 9.53 
                 4.47 
                 91.51 
                 10.03 
               
               
                 AD-203162.1 
                 9.59 
                 2.67 
                 74.89 
                 4.45 
               
               
                 AD-202130.1 
                 9.91 
                 1.83 
                 73.08 
                 13.47 
               
               
                 AD-203169.1 
                 9.94 
                 4.10 
                 65.00 
                 17.66 
               
               
                 AD-202052.1 
                 10.49 
                 1.35 
                 82.10 
                 14.23 
               
               
                 AD-203184.1 
                 11.77 
                 1.68 
                 70.99 
                 14.90 
               
               
                 AD-202492.1 
                 13.12 
                 7.39 
                 97.36 
                 9.93 
               
               
                 AD-202491.1 
                 13.64 
                 3.25 
                 83.92 
                 13.16 
               
               
                 AD-202050.1 
                 13.95 
                 3.30 
                 98.93 
                 8.09 
               
               
                 AD-202197.1 
                 14.19 
                 4.52 
                 85.40 
                 4.49 
               
               
                 AD-202192.1 
                 14.56 
                 4.70 
                 93.25 
                 9.88 
               
               
                 AD-203148.1 
                 15.18 
                 10.67 
                 67.89 
                 6.79 
               
               
                 AD-202499.1 
                 15.88 
                 4.59 
                 87.15 
                 8.82 
               
               
                 AD-202054.1 
                 16.12 
                 4.62 
                 60.58 
                 8.56 
               
               
                 AD-202372.1 
                 17.57 
                 4.27 
                 97.08 
                 17.65 
               
               
                 AD-202796.1 
                 17.61 
                 6.93 
                 99.33 
                 11.62 
               
               
                 AD-206288.1 
                 17.62 
                 2.30 
                 102.90 
                 2.78 
               
               
                 AD-202191.1 
                 18.07 
                 10.80 
                 50.60 
                 9.23 
               
               
                 AD-203177.1 
                 18.83 
                 8.19 
                 81.37 
                 21.78 
               
               
                 AD-202489.1 
                 19.13 
                 4.49 
                 102.86 
                 6.67 
               
               
                 AD-202387.1 
                 19.58 
                 4.55 
                 110.69 
                 13.33 
               
               
                 AD-202399.1 
                 21.07 
                 2.81 
                 101.54 
                 8.60 
               
               
                 AD-202386.1 
                 22.14 
                 10.71 
                 82.79 
                 7.45 
               
               
                 AD-202053.1 
                 22.16 
                 9.75 
                 88.51 
                 8.66 
               
               
                 AD-203166.1 
                 22.83 
                 9.53 
                 97.08 
                 10.70 
               
               
                 AD-202827.1 
                 23.37 
                 5.69 
                 97.37 
                 9.83 
               
               
                 AD-205123.1 
                 23.44 
                 3.98 
                 109.19 
                 22.08 
               
               
                 AD-202490.1 
                 27.49 
                 11.58 
                 93.09 
                 23.32 
               
               
                 AD-202051.1 
                 28.61 
                 9.86 
                 106.66 
                 25.53 
               
               
                 AD-202894.1 
                 30.30 
                 16.53 
                 95.28 
                 5.66 
               
               
                 AD-202494.1 
                 31.84 
                 14.08 
                 112.19 
                 13.26 
               
               
                 AD-202905.1 
                 33.51 
                 10.24 
                 103.04 
                 12.40 
               
               
                 AD-202904.1 
                 36.44 
                 8.37 
                 111.35 
                 12.49 
               
               
                 AD-203151.1 
                 38.01 
                 10.25 
                 116.83 
                 20.09 
               
               
                 AD-202406.1 
                 39.88 
                 12.29 
                 114.12 
                 13.26 
               
               
                 AD-202913.1 
                 42.05 
                 9.55 
                 110.51 
                 4.43 
               
               
                 AD-202277.1 
                 42.85 
                 17.43 
                 109.60 
                 11.58 
               
               
                 AD-202133.1 
                 43.57 
                 12.43 
                 109.00 
                 15.57 
               
               
                 AD-202895.1 
                 51.91 
                 13.91 
                 105.13 
                 8.81 
               
               
                 AD-202398.1 
                 52.06 
                 10.33 
                 107.98 
                 15.80 
               
               
                 AD-202136.1 
                 52.87 
                 8.23 
                 113.49 
                 9.99 
               
               
                 AD-202131.1 
                 55.15 
                 17.03 
                 115.20 
                 12.26 
               
               
                 AD-202393.1 
                 55.30 
                 17.11 
                 117.36 
                 16.07 
               
               
                 AD-201919.1 
                 60.06 
                 20.20 
                 117.56 
                 11.60 
               
               
                 AD-202787.1 
                 60.78 
                 6.44 
                 101.21 
                 24.69 
               
               
                 AD-205414.1 
                 61.10 
                 4.87 
                 99.19 
                 13.48 
               
               
                 AD-205279.1 
                 61.27 
                 4.05 
                 90.54 
                 24.56 
               
               
                 AD-202373.1 
                 61.83 
                 20.15 
                 120.16 
                 20.29 
               
               
                 AD-202198.1 
                 64.73 
                 9.39 
                 107.54 
                 5.70 
               
               
                 AD-202420.1 
                 76.18 
                 8.20 
                 116.98 
                 23.33 
               
               
                 AD-202199.1 
                 82.21 
                 17.17 
                 103.61 
                 2.75 
               
               
                 AD-204885.1 
                 82.62 
                 35.39 
                 115.74 
                 1.39 
               
               
                 AD-205284.1 
                 83.03 
                 24.48 
                 106.27 
                 27.83 
               
               
                 AD-205458.1 
                 83.31 
                 23.23 
                 105.38 
                 10.06 
               
               
                 AD-205413.1 
                 84.64 
                 15.14 
                 114.56 
                 20.06 
               
               
                 AD-206058.1 
                 90.57 
                 29.97 
                 107.33 
                 8.99 
               
               
                 AD-205411.1 
                 94.59 
                 12.06 
                 118.63 
                 28.93 
               
               
                 AD-202407.1 
                 106.96 
                 17.59 
                 113.18 
                 17.86 
               
               
                 AD-205459.1 
                 109.74 
                 31.14 
                 106.34 
                 18.26 
               
               
                 AD-205455.1 
                 111.71 
                 21.62 
                 112.78 
                 7.76 
               
               
                 AD-205264.1 
                 112.90 
                 6.47 
                 112.44 
                 18.05 
               
               
                 AD-204884.1 
                 113.22 
                 11.72 
                 107.09 
                 3.43 
               
               
                 AD-204888.1 
                 119.44 
                 3.35 
                 111.36 
                 12.87 
               
               
                 AD-205270.1 
                 120.85 
                 19.15 
                 105.61 
                 20.53 
               
               
                 AD-204894.1 
                 120.98 
                 20.92 
                 105.19 
                 6.54 
               
               
                 AD-205273.1 
                 122.39 
                 15.30 
                 111.46 
                 11.37 
               
               
                 AD-206218.1 
                 122.51 
                 25.67 
                 112.92 
                 10.34 
               
               
                 AD-202132.1 
                 122.82 
                 23.67 
                 132.29 
                 15.56 
               
               
                 AD-202389.1 
                 122.84 
                 9.76 
                 117.70 
                 6.54 
               
               
                 AD-204944.1 
                 124.59 
                 18.43 
                 107.63 
                 10.53 
               
               
                 AD-205463.1 
                 125.93 
                 19.08 
                 122.92 
                 19.90 
               
               
                 AD-206057.1 
                 126.66 
                 12.62 
                 108.80 
                 11.55 
               
               
                 AD-206281.1 
                 126.70 
                 29.26 
                 117.25 
                 17.44 
               
               
                 AD-206059.1 
                 130.04 
                 18.75 
                 116.48 
                 17.30 
               
               
                 AD-206219.1 
                 132.66 
                 23.95 
                 101.59 
                 18.88 
               
               
                 AD-205227.1 
                 139.82 
                 21.78 
                 123.34 
                 30.07 
               
               
                 AD-204943.1 
                 140.50 
                 32.99 
                 114.02 
                 14.07 
               
               
                 AD-204882.1 
                 144.54 
                 25.34 
                 118.80 
                 16.49 
               
               
                 AD-205214.1 
                 145.54 
                 14.94 
                 110.48 
                 25.47 
               
               
                 AD-206056.1 
                 146.84 
                 18.86 
                 117.44 
                 9.63 
               
               
                 AD-205456.1 
                 149.51 
                 22.58 
                 111.56 
                 10.31 
               
               
                 AD-205464.1 
                 159.65 
                 26.57 
                 119.56 
                 25.63 
               
               
                 AD-205401.1 
                 162.58 
                 43.64 
                 138.49 
                 20.23 
               
               
                 AD-205034.1 
                 169.05 
                 28.31 
                 133.42 
                 32.79 
               
               
                   
               
            
           
         
       
     
     
       
         
           
               
             
               
                 TABLE 5 
               
             
            
               
                   
               
               
                 SERPINF2 Single 10 nM and 0.1 nM Dose Screen in PMH cells 
               
               
                 (values indicate % of message remaining) 
               
            
           
           
               
               
               
               
               
            
               
                   
                 10 nM  
                 10 nM 
                 0.1 nM 
                 0.1 nM 
               
               
                 Duplex ID 
                 Average 
                 STDEV 
                 Average 
                 STDEV 
               
               
                   
               
            
           
           
               
               
               
               
               
            
               
                 AD-203163.1 
                 48.72 
                 12.19 
                 91.98 
                 27.42 
               
               
                 AD-203147.1 
                 106.09 
                 25.50 
                 97.57 
                 27.79 
               
               
                 AD-202129.1 
                 6.92 
                 1.49 
                 55.97 
                 14.69 
               
               
                 AD-203171.1 
                 8.93 
                 2.20 
                 55.64 
                 19.00 
               
               
                 AD-202239.1 
                 24.07 
                 7.42 
                 83.84 
                 23.71 
               
               
                 AD-202137.1 
                 15.87 
                 4.20 
                 151.84 
                 29.04 
               
               
                 AD-202196.1 
                 21.01 
                 3.83 
                 144.43 
                 21.16 
               
               
                 AD-203162.1 
                 64.83 
                 24.12 
                 100.92 
                 12.24 
               
               
                 AD-202130.1 
                 12.64 
                 2.27 
                 83.03 
                 10.31 
               
               
                 AD-203169.1 
                 7.73 
                 2.48 
                 93.44 
                 28.85 
               
               
                 AD-202052.1 
                 23.51 
                 7.49 
                 173.21 
                 50.33 
               
               
                 AD-203184.1 
                 13.41 
                 3.03 
                 175.24 
                 18.56 
               
               
                 AD-202492.1 
                 69.03 
                 9.85 
                 98.06 
                 4.10 
               
               
                 AD-202491.1 
                 80.07 
                 23.91 
                 70.78 
                 11.97 
               
               
                 AD-202050.1 
                 21.86 
                 4.91 
                 128.65 
                 24.69 
               
               
                 AD-202197.1 
                 41.27 
                 14.01 
                 84.85 
                 14.98 
               
               
                 AD-202192.1 
                 19.22 
                 3.81 
                 197.11 
                 40.49 
               
               
                 AD-203148.1 
                 93.44 
                 26.65 
                 84.59 
                 32.55 
               
               
                 AD-202499.1 
                 80.88 
                 22.76 
                 86.53 
                 29.38 
               
               
                 AD-202054.1 
                 6.94 
                 0.85 
                 60.85 
                 15.72 
               
               
                 AD-202372.1 
                 113.61 
                 36.27 
                 105.15 
                 16.68 
               
               
                 AD-202796.1 
                 72.43 
                 5.23 
                 141.69 
                 58.25 
               
               
                 AD-206288.1 
                 5.76 
                 0.82 
                 72.01 
                 42.31 
               
               
                 AD-202191.1 
                 5.68 
                 1.45 
                 71.06 
                 12.31 
               
               
                 AD-203177.1 
                 17.13 
                 2.88 
                 168.42 
                 57.48 
               
               
                 AD-202489.1 
                 163.05 
                 27.91 
                 182.92 
                 31.00 
               
               
                 AD-202387.1 
                 144.06 
                 13.65 
                 178.12 
                 56.65 
               
               
                 AD-202399.1 
                 113.28 
                 23.19 
                 135.14 
                 27.47 
               
               
                 AD-202386.1 
                 142.31 
                 53.78 
                 160.98 
                 75.09 
               
               
                 AD-202053.1 
                 30.41 
                 10.84 
                 153.67 
                 30.69 
               
               
                 AD-203166.1 
                 31.72 
                 2.87 
                 117.70 
                 18.90 
               
               
                 AD-202827.1 
                 173.32 
                 33.42 
                 89.09 
                 39.49 
               
               
                 AD-205123.1 
                 44.75 
                 4.54 
                 90.65 
                 20.52 
               
               
                 AD-202490.1 
                 126.16 
                 29.81 
                 114.40 
                 44.49 
               
               
                 AD-202051.1 
                 27.19 
                 5.06 
                 161.21 
                 68.60 
               
               
                 AD-202894.1 
                 95.28 
                 26.23 
                 83.40 
                 23.60 
               
               
                 AD-202494.1 
                 160.55 
                 28.29 
                 171.80 
                 55.51 
               
               
                 AD-202905.1 
                 144.20 
                 38.76 
                 116.63 
                 34.43 
               
               
                 AD-202904.1 
                 158.48 
                 26.93 
                 152.99 
                 42.92 
               
               
                 AD-203151.1 
                 146.22 
                 7.65 
                 166.88 
                 38.14 
               
               
                 AD-202406.1 
                 14.65 
                 5.23 
                 147.36 
                 43.26 
               
               
                 AD-202913.1 
                 144.16 
                 14.66 
                 129.93 
                 37.09 
               
               
                 AD-202277.1 
                 130.67 
                 28.87 
                 105.89 
                 29.69 
               
               
                 AD-202133.1 
                 86.38 
                 10.26 
                 179.68 
                 48.02 
               
               
                 AD-202895.1 
                 146.53 
                 19.39 
                 145.01 
                 38.87 
               
               
                 AD-202398.1 
                 141.72 
                 20.47 
                 127.39 
                 33.80 
               
               
                 AD-202136.1 
                 114.74 
                 15.28 
                 179.00 
                 42.44 
               
               
                 AD-202131.1 
                 110.12 
                 16.71 
                 185.82 
                 19.51 
               
               
                 AD-202393.1 
                 158.60 
                 17.36 
                 176.78 
                 28.97 
               
               
                 AD-201919.1 
                 143.53 
                 25.59 
                 128.74 
                 53.19 
               
               
                 AD-202787.1 
                 152.60 
                 29.52 
                 196.53 
                 20.75 
               
               
                 AD-205414.1 
                 23.45 
                 3.84 
                 160.02 
                 52.25 
               
               
                 AD-205279.1 
                 26.52 
                 4.52 
                 110.95 
                 14.45 
               
               
                 AD-202373.1 
                 144.10 
                 10.49 
                 202.66 
                 15.52 
               
               
                 AD-202198.1 
                 107.60 
                 5.84 
                 116.19 
                 43.35 
               
               
                 AD-202420.1 
                 128.73 
                 22.47 
                 163.98 
                 19.61 
               
               
                 AD-202199.1 
                 114.74 
                 15.52 
                 124.15 
                 27.26 
               
               
                 AD-204885.1 
                 13.98 
                 2.59 
                 79.21 
                 65.51 
               
               
                 AD-205284.1 
                 56.54 
                 6.90 
                 75.08 
                 17.39 
               
               
                 AD-205458.1 
                 6.72 
                 1.03 
                 73.21 
                 4.23 
               
               
                 AD-205413.1 
                 40.48 
                 5.09 
                 82.62 
                 29.94 
               
               
                 AD-206058.1 
                 80.11 
                 17.91 
                 79.28 
                 19.51 
               
               
                 AD-205411.1 
                 109.03 
                 19.09 
                 76.11 
                 27.41 
               
               
                 AD-202407.1 
                 159.96 
                 29.08 
                 196.07 
                 68.28 
               
               
                 AD-205459.1 
                 16.66 
                 3.49 
                 150.11 
                 48.58 
               
               
                 AD-205455.1 
                 90.23 
                 27.32 
                 89.69 
                 24.06 
               
               
                 AD-205264.1 
                 12.99 
                 4.90 
                 162.72 
                 33.33 
               
               
                 AD-204884.1 
                 10.68 
                 3.20 
                 84.68 
                 59.21 
               
               
                 AD-204888.1 
                 39.30 
                 7.39 
                 130.72 
                 54.24 
               
               
                 AD-205270.1 
                 107.94 
                 8.91 
                 129.03 
                 23.76 
               
               
                 AD-204894.1 
                 7.95 
                 2.61 
                 92.13 
                 34.65 
               
               
                 AD-205273.1 
                 97.91 
                 16.90 
                 119.67 
                 61.70 
               
               
                 AD-206218.1 
                 15.42 
                 3.47 
                 93.12 
                 16.02 
               
               
                 AD-202132.1 
                 138.86 
                 28.87 
                 159.57 
                 51.07 
               
               
                 AD-202389.1 
                 156.09 
                 12.09 
                 170.42 
                 66.34 
               
               
                 AD-204944.1 
                 38.03 
                 13.46 
                 125.02 
                 56.26 
               
               
                 AD-205463.1 
                 24.85 
                 1.51 
                 180.42 
                 68.11 
               
               
                 AD-206057.1 
                 121.80 
                 20.97 
                 150.46 
                 43.19 
               
               
                 AD-206281.1 
                 13.73 
                 1.78 
                 103.54 
                 51.84 
               
               
                 AD-206059.1 
                 30.61 
                 8.96 
                 80.76 
                 31.65 
               
               
                 AD-206219.1 
                 42.02 
                 16.58 
                 148.12 
                 36.61 
               
               
                 AD-205227.1 
                 76.27 
                 5.11 
                 172.29 
                 24.21 
               
               
                 AD-204943.1 
                 12.39 
                 3.99 
                 97.08 
                 38.47 
               
               
                 AD-204882.1 
                 4.83 
                 1.03 
                 79.29 
                 19.40 
               
               
                 AD-205214.1 
                 45.25 
                 7.23 
                 91.38 
                 5.20 
               
               
                 AD-206056.1 
                 68.35 
                 6.16 
                 140.72 
                 8.81 
               
               
                 AD-205456.1 
                 83.91 
                 20.29 
                 160.43 
                 28.61 
               
               
                 AD-205464.1 
                 23.72 
                 1.60 
                 94.79 
                 15.95 
               
               
                 AD-205401.1 
                 96.64 
                 24.08 
                 66.66 
                 20.49 
               
               
                 AD-205034.1 
                 6.68 
                 2.05 
                 75.06 
                 23.42 
               
               
                   
               
            
           
         
       
     
     Example 3. In Vivo Screening of dsRNA Duplexes in Mice Injected with GalNAc-siRNAs Targeting SERPINF2 
     Duplexes of interest, identified from the above in vitro studies, were evaluated in mice. Female wild-type (C57BL/6) mice (n=3 per group) were subcutaneously administered a single 2 mg/kg dose of the siRNA agents shown in  FIG. 1  and listed in Table 6, on day 0. Ten days post dosing, animal liver samples were collected and snap-frozen in liquid nitrogen. Tissue mRNA was extracted and analyzed by the RT-QPCR method. SERPINF2 mRNA levels were compared to housekeeping gene GAPDH. The values were then normalized to the average of PBS vehicle control group. The data were expressed as percent of baseline value, and presented as mean plus standard deviation. The results are listed in Table 7, and shown in  FIG. 2 . 
     The dsRNA duplex AD-329749, listed in Table 8, was selected for further in vivo analysis. Briefly, female wild-type (C57BL/6) mice (n=3 per group) were subcutaneously administered a single 0.3 mg/kg, 3 mg/kg or 10 mg/kg of AD-329749 on day 0. Ten days post dosing, animal liver samples were collected and analyzed for SERPINF2 mRNA levels as described above. The results are shown in  FIG. 3 . 
     
       
         
           
               
             
               
                 TABLE 6 
               
             
            
               
                   
               
               
                  Sense and antisense strand sequences of SERPINF2 dsRNAs for in vivo analysis 
               
            
           
           
               
               
               
               
               
               
               
            
               
                 Duplex 
                 Sense 
                   
                 SEQ 
                 Antisense 
                 Antisense Oligo 
                 SEQ ID 
               
               
                 Name 
                 Oligo Name 
                 Sense Oligo Sequence 
                 ID NO: 
                 Oligo Name 
                 Sequence 
                 NO: 
               
               
                   
               
               
                 AD- 
                   
                 csasgaaaGfgAfUfUfuc 
                 479 
                 A-399626.1 
                 usUfsugaUfgGfGfaaau 
                 487 
               
               
                 202054.2 
                 A-399625.1 
                 ccaucaaaL96 
                   
                   
                 CfcUfuucugscsa 
                   
               
               
                   
               
               
                 AD- 
                   
                 ascscuggCfaAfAfCfau 
                 480 
                 A-399776.1 
                 asUfsuggUfuGfAfugu 
                 488 
               
               
                 202129.2 
                 A-399775.1 
                 caaccaauL96 
                   
                   
                 uUfgCfcagguscsa 
                   
               
               
                   
               
               
                 AD- 
                 A-399899.1 
                 ascscgugUfuGfCfUfuc 
                 481 
                 A-399900.1 
                 usUfsugaGfgAfGfaagc 
                 489 
               
               
                 202191.2 
                   
                 uccucaaaL96 
                   
                   
                 AfaCfacggusgsu 
                   
               
               
                   
               
               
                 AD- 
                 A-401855.1 
                 usgsuaguGfaUfUfUfu 
                 482 
                 A-401856.1 
                 usGfsuggCfaUfAfaaaa 
                 490 
               
               
                 203169.2 
                   
                 uaugccacaL96 
                   
                   
                 UfcAfcuacasasa 
                   
               
               
                   
               
               
                 AD- 
                 A-401859.1 
                 usasgugaUfuUfUfUfa 
                 483 
                 A-401860.1 
                 usAfsgguGfgCfAfuaaa 
                 491 
               
               
                 203171.2 
                   
                 ugccaccuaL96 
                   
                   
                 AfaUfcacuascsa 
                   
               
               
                   
               
               
                 AD- 
                 A-405318.1 
                 gsgsccauGfaUfGfGfcu 
                 484 
                 A-405319.1 
                 usUfsaguAfaAfAfgcca 
                 492 
               
               
                 204882.2 
                   
                 uuuacuaaL96 
                   
                   
                 UfcAfuggccsusg 
                   
               
               
                   
               
               
                 AD- 
                 A-406462.1 
                 usgsgaacGfuGfUfCfcg 
                 485 
                 A-406463.1 
                 usAfsguaCfcUfCfggac 
                 493 
               
               
                 205458.2 
                   
                 agguacuaL96 
                   
                   
                 AfcGfuuccascsu 
                   
               
               
                   
               
               
                 AD- 
                 A-408120.1 
                 gscsucccAfaGfAfCfuc 
                 486 
                 A-408121.1 
                 usUfsacaAfaAfGfaguc 
                 494 
               
               
                 206288.2 
                   
                 uuuuguaaL96 
                   
                   
                 UfuGfggagcsusa 
               
               
                   
               
            
           
         
       
     
     
       
         
           
               
             
               
                 TABLE 7 
               
             
            
               
                   
               
               
                 SERPINF2 Single 2 mg/kg Dose Screen in C57BL/6 mice 
               
            
           
           
               
               
               
            
               
                   
                 % of message 
                   
               
               
                   
                 remaining 
                   
               
               
                 Duplex ID 
                 relative to PBS 
                 STDEV 
               
               
                   
               
            
           
           
               
               
               
            
               
                 PBS 
                 100.04 
                 3.15 
               
               
                 AD-202054.2 
                 75.61 
                 3.08 
               
               
                 AD-202129.2 
                 91.33 
                 6.48 
               
               
                 AD-202191.2 
                 62.40 
                 1.87 
               
               
                 AD-203169.2 
                 41.32 
                 3.66 
               
               
                 AD-203171.2 
                 57.01 
                 3.35 
               
               
                 AD-204882.2 
                 54.36 
                 14.93 
               
               
                 AD-205458.2 
                 82.25 
                 14.80 
               
               
                 AD-206288.2 
                 46.51 
                 4.68 
               
               
                   
               
            
           
         
       
     
     
       
         
           
               
             
               
                 TABLE 8 
               
             
            
               
                   
               
               
                 SERPINF2 Single 0.3 mg/kg, 3 mg/kg and 10 mg/kg Dose Screen in C57BL/6 mice 
               
            
           
           
               
               
               
               
               
               
               
            
               
                   
                   
                   
                   
                 SEQ ID 
                 Unmodified 
                 SEQ ID 
               
               
                 Duplex ID 
                 Oligo Name 
                 strand 
                 Oligo Sequence 
                  NO: 
                 sequence 
                 NO: 
               
               
                   
               
               
                 AD-329749 
                 A-401855 
                 sense 
                 usgsuaguGfaUfUfU 
                 495 
                 UGUAGUGAUUUUU 
                 497 
               
               
                   
                   
                   
                 fuuaugccacaL96 
                   
                 AUGCCACA 
                   
               
               
                   
               
               
                   
                 A-620653 
                 antis 
                 VPusGfsuggCfaUf 
                 496 
                 UGUGGCAUAAAAA 
                 498 
               
               
                   
                   
                   
                 AfaaaaUfcAfcuacas 
                   
                 UCACUACAAA 
                   
               
               
                   
                   
                   
                 asa 
               
               
                   
               
            
           
         
       
     
     Example 4. Knocking Down Hepatic SERPINF2 by GalNAc-siRNA Strategy Expedited Thrombolysis in a Rodent Large-Vein Electrolytic Injury Model 
     Post-Thrombotic Syndrome (PTS) is a chronic complication of deep venous thrombosis (DVT) and has been estimated to affect 40% of DVT patients within two years. Thrombolytic therapy may reduce the burden of PTS. A2AP is a serine protease inhibitor responsible for inactivating plasmin. In this study, a mouse large-vein electrolytic injury model was exploited to evaluate the effect of SERPINF2 suppression on dynamic thrombosis and thrombolysis. 
     C57Bl/6 mice (3-4 months old) were subcutaneously administered a 10 mg/kg dose of AD-203169 on a weekly basis to ensure sustained A2AP mRNA reduction. On day 0, animals were subcutaneously administered a 10 mg/kg dose of AD-203169. On day 10, thrombosis formation was induced through large vein electrolytic injury (3V, 90 s). Five minutes prior to this thrombus induction, rhodamine 6G (0.5 mg/kg) and anti-fibrin antibody (monoclonal 59D8) labeled with Alexa-Fluor-647 (Invitrogen) were injected into the external jugular vein. The femoral vein thrombus site was subsequently imaged with intra-vital microscopy using time-lapse capture over 60 min to record fibrin deposition. (Cooley et al., Murine model of large-vein electrolytic injury induction of thrombosis with slow resolution, Trombosis Research (2015)). 
     Subsequently, wounds were closed and animals recovered from anesthesia Animals were sacrificed on day 11 (24 hrs post injury). Liver tissues were collected (liquid nitrogen). Liquid nitrogen collected liver samples (in grinding jars) can be stored at −80° C. before further analysis. Electrolytic injured veins were also collected. Each harvested vein with thrombus was fixed in 4% buffered formaldehyde, processed with paraffin embedding, and sectioned longitudinally in its entirety for Haemotoxylin and Eosin staining and histo-morphometric volume reconstruction of the residual thrombus and remodeling vein wall. 
     As shown in  FIGS. 4A and 4B  and Table 9, weekly administration of GalNAc-siRNAs targeting A2AP led to &gt;90% target liver mRNA suppression. Twenty-four hours post injury, animals with A2AP knockdown displayed reduced fibrin deposition ( FIG. 4A ), and a significant decrease in thrombus size ( FIG. 4B ). These results demonstrate that knocking down hepatic A2AP accelerated thrombolysis in the mouse electrolytic injury model. Accordingly, knocking down hepatic SERPINF2 accelerated thrombolysis in mouse electrolytic injury model and, thus, demonstrates threat targeting SERPINF2 with a dsRNA is a useful therapeutic for post thrombotic syndrome. 
     
       
         
           
               
             
               
                 TABLE 9 
               
             
            
               
                   
               
               
                 Histomorphometric analysis 
               
               
                 of the vein with thrombus 
               
            
           
           
               
               
               
            
               
                   
                   
                 A2AP- 
               
               
                   
                 PBS 
                 siRNA 
               
               
                   
                   
               
            
           
           
               
               
               
            
               
                   
                 0.193655 
                 0.104445 
               
               
                   
                 0.369339 
                 0.037008 
               
               
                   
                 0.220671 
                 0.062907 
               
               
                   
                 0.15794 
                 0.042792 
               
               
                   
                 0.292683 
                 0.135604 
               
               
                   
                 0.409563 
                 0.066414 
               
               
                   
                 0.189599 
                 0.136267 
               
               
                   
                 0.377682 
               
               
                   
                   
               
            
           
         
       
     
     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.