Patent Publication Number: US-2021171956-A1

Title: GLUCOKINASE (GCK) iRNA COMPOSITIONS AND METHODS OF USE THEREOF

Description:
RELATED APPLICATIONS 
     This application is a continuation of U.S. patent application Ser. No. 15/849,776, filed on Dec. 21, 2017, which is a 35 § U.S.C. 111(a) continuation application which claims the benefit of priority to PCT/US2016/038616, filed on Jun. 22, 2016, which claims the benefit of priority of U.S. Provisional Patent Application No. 62/183,413, filed on Jun. 23, 2015, U.S. Provisional Patent Application No. 62/200,207, filed on Aug. 3, 2015, and U.S. Provisional Patent Application No. 62/260,876, filed on Nov. 30, 2015. The entire contents of each of the foregoing applications are hereby 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 Oct. 1, 2020, is named 121301-03705_SL.txt and is 523,767 bytes in size. 
     BACKGROUND OF THE INVENTION 
     Hypoglycemia is a biochemical symptom indicating the presence of an underlying disease or disorder. Since glucose is the fundamental energy currency of the cell, diseases and disorders that affect its availability or use can cause hypoglycemia. 
     The body normally defends against hypoglycemia by decreasing insulin secretion and increasing glucagon, epinephrine, growth hormone, and cortisol secretion. These hormonal changes combine to increase hepatic glucose production, increase alternative fuel availability, and decrease glucose use. The increase in hepatic glucose production is initially caused by the breakdown of liver glycogen stores resulting from lower insulin levels and increased glucagon levels. When glycogen stores become depleted and protein breakdown increases because of increased cortisol levels, hepatic gluconeogenesis replaces glycogenolysis as the primary source of glucose production. Decreased use of peripheral glucose occurs initially because of a fall in insulin levels and later because of increases in epinephrine, cortisol, and growth hormone levels. 
     All of these events increase lipolysis and plasma free fatty acid levels, which are then available as an alternative fuel source and act to competitively inhibit glucose use. Plasma free fatty acids also stimulate glucose production. Hypoglycemia occurs when one or more of these counterregulatory mechanisms fail because of, for example, the overuse of glucose (as in hyperinsulinism) or the underproduction of glucose (as in a glycogen-storage diseases). 
     GSD is an inherited genetic disorder due to an absence or deficiency of one of the enzymes responsible for making or breaking down glycogen in the body. This enzyme deficiency causes either abnormal tissue concentrations of glycogen or incorrectly or abnormally formed glycogen and patients with glycogen storage diseases (GSD) may also have low blood glucose levels. GSD occurs in about one of 50,000 to 100,000 births. Some patients might die before diagnosis, while severe infantile forms and some milder forms might go unrecognized. Symptoms of GSD vary based on the enzyme that is missing and usually result from the buildup of glycogen or from the inability to produce glucose when needed. Because GSD occurs mainly in muscles and the liver, those areas show the most symptoms, such as, poor growth, muscle cramps, low blood sugar, enlarged liver, swollen belly, and abnormal blood chemistry. 
     Current treatments for glycogen storage diseases have been very limited and mainly focused on correcting hypoglycemia and other metabolic disturbances through dietary control, such as, by using a modified form of cornstarch, or a high-protein diet. 
     Accordingly, there is a need in the art for alternative treatments for subjects having a glycogen storage disease (GSD), e.g., type Ia GSD, which are independent of the underlying molecular defects. 
     SUMMARY OF THE INVENTION 
     The present invention provides iRNA compositions which effect the RNA-induced silencing complex (RISC)-mediated cleavage of RNA transcripts of a glucokinase (GCK) gene. The GCK gene may be within a cell, e.g., a cell within a subject, such as a human. The present invention also provides methods of using the iRNA compositions of the invention for inhibiting the expression of a GCK gene and/or for treating a subject who would benefit from inhibiting or reducing the expression of a GCK gene, e.g., a subject suffering or prone to suffering from a glycogen storage disease (GSD), or one or more signs or symptoms of GSD, such as hypoglycemia, lactic acidosis, hyperuricemia, hyperlipidemia, hepatomegaly, kidney disease as a result if glycogen accumulation, hunger, jitteriness, lethargy, apnea, seizures, diaphoresis, confusion, headaches, dizziness, unusual mood or behavior changes, loss of consciousness, coma, muscle cramps, bleeding diathesis, short stature, osteoporosis, delayed puberty, gout, renal disease, systemic hypertension, pulmonary hypertension, hepatic adenomas, pancreatitis, anemia, vitamin D deficiency, polycystic ovaries, irregular menstrual cycles, menorrhagia, and/or eruptive xanthomata. 
     GCK in the liver of a subject is the rate limiting enzyme for both glucose uptake and glycogen synthesis. Furthermore, the liver is a major site for regulation of whole body glucose metabolism. Therefore, by inhibiting expression of a glucokinase gene, e.g., a glucokinase gene in the liver of a subject, with a double stranded RNAi agent of the invention, e.g., an RNAi agent that contains and/or is coupled to a ligand, e.g., GalNAc3, that directs the RNAi agent to a site of interest, e.g., the liver, hepatic glucose uptake and storage will decrease, thus, inhibiting hypoglycemia, e.g., fasting hypoglycemia, in subjects suffering or prone to suffering from a glycogen storage disease (GSD), e.g., type Ia GSD, independent of the underlying cause. Furthermore, by inhibiting expression of a glucokinase gene, e.g., a glucokinase gene in the liver of a subject, with a double stranded RNAi agent of the invention, insulin resistance in the liver is selectively induced without altering insulin clearance in order to maintain safe glucose levels in subjects suffering or prone to suffering from a glycogen storage disease (GSD), e.g., type Ia GSD. In addition, by inhibiting expression of a glucokinase gene, e.g., a glucokinase gene in the liver of a subject having a glycogen storage disease (GSD), e.g., type Ia GSD, with a double stranded RNAi agent of the invention, hepatic glucose uptake and storage will decrease, thus, inhibiting hypoglycemia, e.g., fasting hypoglycemia, inhibit hepatic glycogen storage to inhibit hepatomegaly, reduce lipid abnormalities, lactic acidosis and/or hyperuricemia. 
     Accordingly, in one aspect, the present invention provides double stranded ribonucleic acids (RNAi) agents for inhibiting expression of a glucokinase (GCK) gene. The double stranded RNAi agents comprise 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:15. 
     In another aspect, the invention provides double stranded ribonucleic acids (RNAi) agents for inhibiting expression of glucokinase (GCK), wherein the double stranded RNAi agents comprises a sense strand and an antisense strand, the antisense strand comprising a region of complementarity which comprises at least 15 contiguous nucleotides differing no more than 3 nucleotides from any one of the antisense sequences listed in any one of Tables 2, 3, 6, and 7. 
     In certain embodiments, the double stranded RNAi agent comprises at least one modified nucleotide. In one embodiment, the modified nucleotide comprises a 2′-O-methyl modified nucleotide. In another embodiment, the modified nucleotide comprises a 2′-fluoro modified nucleotide. In one embodiment, the modified nucleotide comprises a 3′-terminal deoxy-thymine (dT) nucleotide. In another embodiment, the modified nucleotide comprises a short sequence of deoxy-thymine (dT) nucleotides. 
     In certain embodiments, the dsRNA comprises no more than 4 (i.e., 4, 3, 2, 1, or 0) unmodified nucleotides in the sense strand. In certain embodiments, the dsRNA comprises no more than 4 (i.e., 4, 3, 2, 1, or 0) unmodified nucleotides in the antisense strand. In certain embodiments, the double stranded RNAi agent comprises no more than 4 (i.e., 4, 3, 2, 1, or 0) unmodified nucleotides in both the sense strand and the antisense strand. In certain embodiments, the double stranded RNAi agent comprises all modified nucleotides in the sense strand. In certain embodiments, the dsRNA comprises all modified nucleotides in the antisense strand. In certain embodiments, the double stranded RNAi agent comprises all modified nucleotides in both the sense strand and the antisense strand. 
     In one aspect, the present invention provides double stranded ribonucleic acid (RNAi) agents for inhibiting expression of glucokinase (GCK), wherein the double stranded RNAi agent comprises a sense strand and an antisense strand forming a double stranded region, 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:15, wherein substantially all of the nucleotides of the sense strand and substantially all of the nucleotides of the antisense strand are modified nucleotides, and wherein the sense strand is conjugated to a ligand attached at the 3′-terminus. 
     In certain embodiments, at least one of the modified nucleotides is selected from the group consisting of 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 5′-phosphorothioate group, a nucleotide comprising a 5′-methylphosphonate group, a nucleotide comprising a 5′ phosphate or 5′ phosphate mimic, a nucleotide comprising vinyl phosphate, a nucleotide comprising adenosine-glycol nucleic acid (GNA), a nucleotide comprising thymidine-glycol nucleic acid (GNA)S-Isomer, a nucleotide comprising 2-hydroxymethyl-tetrahydrofurane-5-phosphate, a nucleotide comprising 2′-deoxythymidine-3′phosphate, a nucleotide comprising 2′-deoxyguanosine-3′-phosphate, and a terminal nucleotide linked to a cholesteryl derivative or a dodecanoic acid bisdecylamide group. 
     In certain embodiments, the modified nucleotides is/are independently selected from the group consisting of a 2′-O-methyl modified nucleotide, a nucleotide comprising a 5′-phosphorothioate group, a 3′-terminal deoxy-thymine (dT) nucleotide, and a terminal nucleotide linked to a cholesteryl derivative or a dodecanoic acid bisdecylamide group. In certain embodiments, the modified nucleotide is selected from the group consisting of a 2′-deoxy-2′-fluoro modified nucleotide, a 2′-deoxy-modified nucleotide, a 3′-terminal deoxy-thymine (dT) nucleotide, a locked nucleotide, an abasic nucleotide, a 2′-amino-modified nucleotide, a 2′-alkyl-modified nucleotide, a morpholino nucleotide, a phosphoramidate, and a non-natural base comprising nucleotide. 
     In certain embodiments, the double stranded RNAi agent comprises a region of complementarity at least 17 nucleotides in length. In certain embodiments, the double stranded RNAi agent comprises a region of complementarity 19-23 nucleotides in length. In certain embodiments, the double stranded RNAi agent comprises a region of complementarity 19-21 nucleotides in length. In certain embodiments, the double stranded RNAi agent comprises a region of complementarity is 19 nucleotides in length. In certain embodiments, the double stranded RNAi agent comprises a region of complementarity is 21 nucleotides in length. 
     In certain embodiments, each strand of the double stranded RNAi agent is no more than 30 nucleotides in length. In certain embodiments, the double stranded RNAi agent is at least 15 nucleotides in length. 
     In some embodiments, at least one strand of the dsRNA agent comprises a 3′ overhang of at least 1 nucleotide, e.g., at least one strand comprises a 3′ overhang of at least 2 nucleotides, e.g., 2, 3, 4, 5, 6, 7, 9, 10, 11, 12, 13, 14, or 15 nucleotides. In some embodiments, at least one strand of the RNAi agent comprises a 5′ overhang of at least 1 nucleotide. In certain embodiments, at least one strand comprises a 5′ overhang of at least 2 nucleotides, e.g., 2, 3, 4, 5, 6, 7, 9, 10, 11, 12, 13, 14, or 15 nucleotides. In some embodiments, both the 3′ and the 5′ end of one strand of the RNAi agent comprise an overhang of at least 1 nucleotide. In some embodiments, at least one strand comprises a 3′ overhang of at least 2 nucleotides. 
     In certain embodiments, the double stranded RNAi agent further comprises a ligand. In certain embodiments, the ligand is conjugated to the 3′ end of the sense strand of the double stranded RNAi agent. In certain embodiments, the ligand is an N-acetylgalactosamine (GalNAc) derivative. 
     In certain embodiments, the ligand is 
     
       
         
         
             
             
         
       
     
     In certain embodiments, the wherein the double stranded RNAi agent is conjugated to the ligand as shown in the following schematic 
     
       
         
         
             
             
         
       
     
     and, wherein X is O or S. 
     In certain embodiments, the ligand is a cholesterol. 
     In certain embodiments, the ligand is one or more GalNAc derivatives attached through a monovalent, a bivalent, or a trivalent branched linker. The ligand may be conjugated to the 3′ end of the sense strand of the polynucleotide agent, the 5′ end of the sense strand of the polynucleotide agent, the 3′ end of the antisense strand of the polynucleotide agent, the 5′ end of the antisense strand of the polynucleotide agent. 
     In some embodiments, the polynucleotide agents of the invention comprise a plurality, e.g., 2, 3, 4, 5, or 6, of GalNAc, each independently attached to a plurality of nucleotides of the polynucleotide agent through a plurality of monovalent linkers. 
     In one embodiment, the agents inhibit the expression of GCK and the sense strand and the antisense strand comprise nucleotide sequences selected from the group consisting of the nucleotide sequences of any one of the agents listed in any one of Tables 2, 3, 6, and 7. 
     The invention provides cells containing the double stranded RNAi agents provided herein. 
     In another aspect, the invention provides pharmaceutical composition for inhibiting expression of a glucokinase (GCK) gene comprising the double stranded RNAi agent provided herein. In certain embodiments, the pharmaceutical compositions further comprise a lipid formulation. 
     The invention also provides methods of inhibiting expression of a glucokinase (GCK) gene in a cell. The methods include contacting the cell with a double stranded RNAi agent described herein; and maintaining the cell produced for a time sufficient to obtain degradation of the mRNA transcript of a GCK gene, thereby inhibiting expression of the GCK 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 human subject suffers from a disease or disorder that would benefit from reduction in GCK expression, such as a glycogen storage disease (GSD), e.g., type Ia GSD. In certain embodiments, the GCK expression is inhibited by at least about 5%, 10%, 15%, 20%, 25%, 30%, about 40%, about 50%, about 60%, about 70%, about 80%, about 90%, about 95%, about 98% or about 100%. 
     The invention further provides methods of treating a subject having a disorder that would benefit from reduction in expression of a glucokinase (GCK) gene. The methods include administering to the subject a therapeutically effective amount of any of the double stranded RNAi agents provided herein, thereby treating the subject, such as a disease or disorder associated with a glycogen storage disease (GSD), e.g., type Ia GSD. 
     In one aspect, the present invention provides methods of preventing at least one symptom in a subject having a disease or disorder that would benefit from reduction in expression of a glucokinase (GCK) gene. The methods include administering to the subject a prophylactically effective amount of a double stranded RNAi agent described herein, thereby preventing at least one symptom in the subject having a disorder that would benefit from reduction in GCK expression. 
     In certain embodiments, administration of the double stranded RNAi agent to the subject causes an increase in one or more blood glucose and/or a decrease in GCK protein accumulation. In certain embodiments, administration of the double stranded RNAi agent to the subject, e.g., a subject having a glycogen storage disease (GSD), e.g., type Ia GSD, causes a decrease in one or more signs of a glycogen storage disease (GSD), e.g., type Ia GSD, e.g., hypoglycemia, lactic acidosis, hyperuricemia, hyperlipidemia, hepatomegaly, kidney disease as a result of glycogen accumulation, hunger, jitteriness, lethargy, apnea, seizures, diaphoresis, confusion, headaches, dizziness, unusual mood or behavior changes, loss of consciousness, coma, muscle cramps, bleeding diathesis, short stauture, osteoporosis, delayed puberty, gout, renal disease, systemic hypertension, pulmonary hypertension, hepatic adenomas, pancreatitis, anemia, vitamin D deficiency, polycystic ovaries, irregular menstrual cycles, menorrhagia, and/or eruptive xanthomata. 
     The invention also provides methods of inhibiting the expression of a glucokinase (GCK) gene in a subject. The methods include method administering to the subject a therapeutically effective amount of any of the double stranded RNAi agents provided herein or a pharmaceutical composition comprising any of the double stranded RNAi agents provided herein, thereby inhibiting the expression of GCK in the subject. 
     In one aspect, the present invention provides methods of increasing the blood glucose levels, e.g., fasting blood glucose levels, in a subject having a disease or disorder that would benefit from reduction in GCK expression. The methods include administering to the subject a therapeutically effective amount of any of the double stranded RNAi agents provided herein or a pharmaceutical composition comprising any of the double stranded RNAi agents provided herein, thereby increasing the blood glucose levels in the subject. 
     In another aspect, the present invention provides methods of decreasing plasma lactate levels in a subject having a disease or disorder that would benefit from reduction in GCK expression. The methods include administering to the subject a therapeutically effective amount of any of the double stranded RNAi agents provided herein or a pharmaceutical composition comprising any of the double stranded RNAi agents provided herein, thereby decreasing the blood lactate levels in the subject. 
     In yet another aspect, the present invention provides methods of decreasing plasma uric acid levels in a subject having a disease or disorder that would benefit from reduction in GCK expression. The methods include administering to the subject a therapeutically effective amount of any of the double stranded RNAi agents provided herein or a pharmaceutical composition comprising any of the double stranded RNAi agents provided herein, thereby decreasing the plasma uric acidlevels in the subject. 
     In one aspect, the present invention provides methods of decreasing plasma triglyceride levels in a subject having a disease or disorder that would benefit from reduction in GCK expression. The methods include administering to the subject a therapeutically effective amount of any of the double stranded RNAi agents provided herein or a pharmaceutical composition comprising any of the double stranded RNAi agents provided herein, thereby decreasing plasma triglyceride levels in the subject. 
     In another aspect, the present invention provides methods of decreasing total plasma cholesterol levels in a subject having a disease or disorder that would benefit from reduction in GCK expression. The methods include administering to the subject a therapeutically effective amount of any of the double stranded RNAi agents provided herein or a pharmaceutical composition comprising any of the double stranded RNAi agents provided herein, thereby decreasing the total plasma cholesterol levels in said subject. 
     In one aspect, the present invention provides methods of decreasing hepatomegaly in a subject having a disease or disorder that would benefit from reduction in GCK expression. The methods include administering to the subject a therapeutically effective amount of any of the double stranded RNAi agents provided herein or a pharmaceutical composition comprising any of the double stranded RNAi agents provided herein, thereby decreasing hepatomegaly in the subject. 
     In certain embodiments, the dsRNA agent is administered to the subject at a dose of about 0.01 mg/kg to about 10 mg/kg or about 0.5 mg/kg to about 50 mg/kg. In certain embodiments, the dsRNA agent is administered about once per month, once every other two months, or once a quarter (i.e., once every three months), for example, at a dose of about 0.1 mg/kg to about 5.0 mg/kg. 
     In certain embodiments, the double stranded RNA agent is administered to the subject once a week. In certain embodiments, the dsRNA agent is administered to the subject once a month. In certain embodiments, the dsRNA agent is administered once per quarter (i.e., every three months). 
     In some embodiments, the methods of the invention further include administering an additional therapeutic to the subject. In one embodiment, the additional therepaeutic is a sodium-glucose co-transporter 2 (SGLT2) inhibitor, e.g., Dapagliflozin, Canagliflozin, Ipragliflozin (ASP-1941), Tofogliflozin, Empagliflozin, Sergliflozin etabonate, Remogliflozin etabonate (BHVO91009), and Ertugliflozin (PF-04971729/MK-8835. 
     In yet another aspect, the invention provides kits for performing the methods of the invention. In one aspect, the invention provides a kit for performing a method for inhibiting expression of a glucokinase (GCK) gene in a cell by contacting a cell with a double stranded RNAi agent in an amount effective to inhibit expression of the GCK in the cell. The kit comprises an RNAi agent and instructions for use and, optionally, means for administering the RNAi agent to a subject. 
    
    
     DETAILED DESCRIPTION OF THE INVENTION 
     The present invention provides iRNA compositions, which affect the RNA-induced silencing complex (RISC)-mediated cleavage of RNA transcripts of a glucokinase (GCK) gene. The GCK gene may be within a cell, e.g., a cell within a subject, such as a human. The present invention also provides methods of using the iRNA compositions of the invention for inhibiting the expression of a glucokinase (GCK) gene and/or for treating a subject having a disorder that would benefit from inhibiting or reducing the expression of a GCK gene, such as a glycogen storage disease (GSD), e.g., type Ia GSD, and one or more of the signs or symptoms associated therewith. 
     The iRNAs of the invention include an RNA strand (the antisense strand) having a region which is about 30 nucleotides or less in length, e.g., 15-30, 15-29, 15-28, 15-27, 15-26, 15-25, 15-24, 15-23, 15-22, 15-21, 15-20, 15-19, 15-18, 15-17, 18-30, 18-29, 18-28, 18-27, 18-26, 18-25, 18-24, 18-23, 18-22, 18-21, 18-20, 19-30, 19-29, 19-28, 19-27, 19-26, 19-25, 19-24, 19-23, 19-22, 19-21, 19-20, 20-30, 20-29, 20-28, 20-27, 20-26, 20-25, 20-24, 20-23, 20-22, 20-21, 21-30, 21-29, 21-28, 21-27, 21-26, 21-25, 21-24, 21-23, or 21-22 nucleotides in length, which region is substantially complementary to at least part of an mRNA transcript of a GCK gene. 
     In certain embodiments, the iRNAs of the invention include an RNA strand (the antisense strand) which can include longer lengths, for example up to 66 nucleotides, 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 GCK gene. These iRNAs with the longer length antisense strands 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 these iRNAs enables the targeted degradation of mRNAs of a GCK gene in mammals. Very low dosages of GCK iRNAs, in particular, can specifically and efficiently mediate RNA interference (RNAi), resulting in significant inhibition of expression of a GCK gene. Using cell-based assays, the present inventors have demonstrated that iRNAs targeting GCK can mediate RNAi, resulting in significant inhibition of expression of a GCK gene. Thus, methods and compositions including these iRNAs are useful for treating a subject who would benefit by a reduction in the levels and/or activity of a GCK protein, such as a subject having a glycogen storage disease (GSD), e.g., type Ia GSD. 
     The following detailed description discloses how to make and use compositions containing iRNAs to inhibit the expression of a GCK gene, as well as compositions and methods for treating subjects having diseases and disorders that would benefit from inhibition and/or reduction of the expression of this gene. 
     I. Definitions 
     In order that the present invention may be more readily understood, certain terms are first defined. In addition, it should be noted that whenever a value or range of values of a parameter are recited, it is intended that values and ranges intermediate to the recited values are also intended to be part of this invention. 
     The articles “a” and “an” are used herein to refer to one or to more than one (i.e., to at least one) of the grammatical object of the article. By way of example, “an element” means one element or more than one element, e.g., a plurality of elements. 
     The term “including” is used herein to mean, and is used interchangeably with, the phrase “including but not limited to”. The term “or” is used herein to mean, and is used interchangeably with, the term “and/or,” unless context clearly indicates otherwise. 
     The term “about” is used herein to mean within the typical ranges of tolerances in the art. For example, “about” can be understood as within 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 terms “GCK,” “glucokinase,” “hexokinase D,” and “hexokinase 4” refer to an enzyme that facilitates phosphorylation of glucose to glucose-6-phosphate having an amino acid sequence from any vertebrate or mammalian source, including, but not limited to, human, bovine, chicken, rodent, mouse, rat, porcine, ovine, primate, monkey, and guinea pig, unless specified otherwise. The term also refers to fragments and variants of native GCK that maintain at least one in vivo or in vitro activity of a native GCK. The term encompasses full-length unprocessed precursor forms of GCK as well as mature forms resulting from, e.g., post-translational processing. 
     The sequence of a human GCK mRNA transcript (transcript variant 2) can be found at, for example, GenBank Accession No. GI: 15967158 (NM_033507; NCBI GenelD: 2645; SEQ ID NO:1). The sequence of another human GCK mRNA transcript (transcript variant 1) can be found at, for example, GenBank Accession No. GI: 167621407 (NM_000162); SEQ ID NO:2). The sequence of yet another human GCK mRNA transcript (transcript variant 3) can be found at, for example, GenBank Accession No. GI: 15967160 (NM_033508); SEQ ID NO:3). The predicted sequence of a rhesus GCK mRNA transcript (transcript variant ×1) can be found at, for example, GenBank Accession No. GI: 544420246 (XM_005549685; SEQ ID NO:4). The predicted sequence of another rhesus GCK mRNA transcript (transcript variant ×2) can be found at, for example, GenBank Accession No. GI: 544420248 (XM_005549686; SEQ ID NO:5). The predicted sequence of yet another rhesus GCK mRNA transcript (transcript variant ×3) can be found at, for example, GenBank Accession No. GI: 544420250 (XM_005549687; SEQ ID NO:6). The predicted sequence of another rhesus GCK mRNA transcript (transcript variant ×4) can be found at, for example, GenBank Accession No. GI: 544420252 (XM_005549688; SEQ ID NO:7). The sequence of a mouse GCK mRNA transcript (transcript variant 1)can be found at, for example, GenBank Accession No. GI: 565671706 (NM_010292; SEQ ID NO:8). The sequence of another mouse GCK mRNA transcript (transcript variant 2)can be found at, for example, GenBank Accession No. GI: 565671714 (NM_001287386; SEQ ID NO:9). The sequence of a rat GCK mRNA transcript (transcript variant 2) can be found at, for example, GenBank Accession No. GI: 399220372 (NM_012565; SEQ ID NO:10). The sequence of another rat GCK mRNA transcript (transcript variant 1) can be found at, for example, GenBank Accession No. GI: 399220370 (NM_001270849; SEQ ID NO:11). The sequence of yet another rat GCK mRNA transcript (transcript variant 3) can be found at, for example, GenBank Accession No. GI: 399220373 (NM_001270850; SEQ ID NO:12). Additional examples of GCK mRNA sequences are readily available using publicly available databases, e.g., GenBank, UniProt, and OMIM. 
     As used herein, “target sequence” refers to a contiguous portion of the nucleotide sequence of an mRNA molecule formed during the transcription of a GCK gene, including mRNA that is a product of RNA processing of a primary transcription product. In one embodiment, the target portion of the sequence will be at least long enough to serve as a substrate for iRNA-directed cleavage at or near that portion of the nucleotide sequence of an mRNA molecule formed during the transcription of a GCK gene. 
     The target sequence may be from about 9-36 nucleotides in length, e.g., about 15-30 nucleotides in length. For example, the target sequence can be from about 15-30 nucleotides, 15-29, 15-28, 15-27, 15-26, 15-25, 15-24, 15-23, 15-22, 15-21, 15-20, 15-19, 15-18, 15-17, 18-30, 18-29, 18-28, 18-27, 18-26, 18-25, 18-24, 18-23, 18-22, 18-21, 18-20, 19-30, 19-29, 19-28, 19-27, 19-26, 19-25, 19-24, 19-23, 19-22, 19-21, 19-20, 20-30, 20-29, 20-28, 20-27, 20-26, 20-25, 20-24, 20-23, 20-22, 20-21, 21-30, 21-29, 21-28, 21-27, 21-26, 21-25, 21-24, 21-23, or 21-22 nucleotides in length. 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. 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,” and “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 GCK 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 RNAi that interacts with a target RNA sequence, e.g., a GCK 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 double stranded short interfering RNAs (siRNAs) comprising a sense strand and an antisense strand by a Type III endonuclease known as Dicer (Sharp et al. (2001)  Genes Dev.  15:485). Dicer, a ribonuclease-III-like enzyme, processes these dsRNA into 19-23 base pair short interfering RNAs with characteristic two base 3′ overhangs (Bernstein, et al., (2001)  Nature  409:363). These siRNAs are then incorporated into an RNA-induced silencing complex (RISC) where one or more helicases unwind the siRNA duplex, enabling the complementary antisense strand to guide target recognition (Nykanen, et al., (2001)  Cell  107:309). Upon binding to the appropriate target mRNA, one or more endonucleases within the RISC cleave the target to induce silencing (Elbashir, et al., (2001)  Genes Dev.  15:188). Thus, in one aspect the invention relates to a single stranded RNA (ssRNA) (the antisense strand of an siRNA duplex) generated within a cell and which promotes the formation of a RISC complex to effect silencing of the target gene, i.e., a GCK gene. Accordingly, the term “siRNA” is also used herein to refer to an RNAi as described above. 
     In another embodiment, the RNAi agent may be a single-stranded RNA that is introduced into a cell or organism to inhibit a target mRNA. Single-stranded RNAi agents bind to the RISC endonuclease, Argonaute 2, which then cleaves the target mRNA. The single-stranded siRNAs are generally 15-30 nucleotides and are chemically modified. The design and testing of single-stranded RNAs are described in U.S. Pat. No. 8,101,348 and in Lima et al., (2012)  Cell  150:883-894, the entire contents of each of which are hereby incorporated herein by reference. Any of the antisense nucleotide sequences described herein may be used as a single-stranded siRNA as described herein or as chemically modified by the methods described in Lima et al., (2012)  Cell  150:883-894. 
     In another embodiment, an “iRNA” for use in the compositions and methods of the invention is a double stranded RNA and is referred to herein as a “double stranded RNAi agent,” “double stranded RNA (dsRNA) molecule,” “dsRNA agent,” or “dsRNA”. The term “dsRNA”, refers to a complex of ribonucleic acid molecules, having a duplex structure comprising two anti-parallel and substantially complementary nucleic acid strands, referred to as having “sense” and “antisense” orientations with respect to a target RNA, i.e., a GCK 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. 
     The majority of nucleotides of each strand of a dsRNA molecule may be ribonucleotides, but as described in detail herein, each or both strands can also include one or more non-ribonucleotides, e.g., a deoxyribonucleotide and/or a modified nucleotide. In addition, as used in this specification, an “RNAi agent” may include ribonucleotides with chemical modifications; an RNAi agent may include substantial modifications at multiple nucleotides. As used herein, the term “modified nucleotide” refers to a nucleotide having, independently, a modified sugar moiety, a modified internucleotide linkage, and/or modified nucleobase. Thus, the term modified nucleotide encompasses substitutions, additions or removal of, e.g., a functional group or atom, to internucleoside linkages, sugar moieties, or nucleobases. The modifications suitable for use in the agents of the 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 “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 9 to 36 base pairs in length, e.g., about 15-30 base pairs in length, for example, about 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, or 36 base pairs in length, such as about 15-30, 15-29, 15-28, 15-27, 15-26, 15-25, 15-24, 15-23, 15-22, 15-21, 15-20, 15-19, 15-18, 15-17, 18-30, 18-29, 18-28, 18-27, 18-26, 18-25, 18-24, 18-23, 18-22, 18-21, 18-20, 19-30, 19-29, 19-28, 19-27, 19-26, 19-25, 19-24, 19-23, 19-22, 19-21, 19-20, 20-30, 20-29, 20-28, 20-27, 20-26, 20-25, 20-24, 20-23, 20-22, 20-21, 21-30, 21-29, 21-28, 21-27, 21-26, 21-25, 21-24, 21-23, or 21-22 base pairs in length. Ranges and lengths intermediate to the above recited ranges and lengths are also contemplated to be part of the invention. 
     The two strands forming the duplex structure may be different portions of one larger RNA molecule, or they may be separate RNA molecules. Where the two strands are part of one larger molecule, and therefore are connected by an uninterrupted chain of nucleotides between the 3′-end of one strand and the 5′-end of the respective other strand forming the duplex structure, the connecting RNA chain is referred to as a “hairpin loop.” A hairpin loop can comprise at least one unpaired nucleotide. In some embodiments, the hairpin loop can comprise at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, at least 20, at least 23, or more unpaired nucleotides. 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, but can be covalently connected. Where the two strands are connected covalently by means other than an uninterrupted chain of nucleotides between the 3′-end of one strand and the 5′-end of the respective other strand forming the duplex structure, the connecting structure is referred to as a “linker.” The RNA strands may have the same or a different number of nucleotides. The maximum number of base pairs is the number of nucleotides in the shortest strand of the dsRNA minus any overhangs that are present in the duplex. In addition to the duplex structure, an RNAi may comprise one or more nucleotide overhangs. 
     In one embodiment, an RNAi agent of the invention is a dsRNA, each strand of which comprises 19-23 nucleotides, that interacts with a target RNA sequence, e.g., a GCK gene, without wishing to be bound by theory, long double stranded RNA introduced into cells is broken down into siRNA by a Type III endonuclease known as Dicer (Sharp et al. (2001)  Genes Dev.  15:485). Dicer, a ribonuclease-III-like enzyme, processes the dsRNA into 19-23 base pair short interfering RNAs with characteristic two base 3′ overhangs (Bernstein, et al., (2001)  Nature  409:363). The siRNAs are then incorporated into an RNA-induced silencing complex (RISC) where one or more helicases unwind the siRNA duplex, enabling the complementary antisense strand to guide target recognition (Nykanen, et al., (2001)  Cell  107:309). Upon binding to the appropriate target mRNA, one or more endonucleases within the RISC cleave the target to induce silencing (Elbashir, et al., (2001)  Genes Dev.  15:188). In one embodiment, an RNAi agent of the invention is a dsRNA of 24-30 nucleotides that interacts with a target RNA sequence, e.g., a prolyl hydroxylase domain-containing gene, i.e., a PHD1 target mRNA sequence, a PHD2 target mRNA sequence, or a PHD3 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 an iRNA, e.g., a dsRNA. For example, when a 3′-end of one strand of a dsRNA extends beyond the 5′-end of the other strand, or vice versa, there is a nucleotide overhang. A dsRNA can comprise an overhang of at least one nucleotide; alternatively the overhang can comprise at least two nucleotides, at least three nucleotides, at least four nucleotides, at least five nucleotides or more. A nucleotide overhang can comprise or consist of a nucleotide/nucleoside analog, including a deoxynucleotide/nucleoside. The overhang(s) can be on the sense strand, the antisense strand or any combination thereof. Furthermore, the nucleotide(s) of an overhang can be present on the 5′-end, 3′-end or both ends of either an antisense or sense strand of a dsRNA. 
     In one embodiment of the dsRNA, at least one strand comprises a 3′ overhang of at least 1 nucleotide. In another embodiment, at least one strand comprises a 3′ overhang of at least 2 nucleotides, e.g., 2, 3, 4, 5, 6, 7, 9, 10, 11, 12, 13, 14, or 15 nucleotides. In other embodiments, at least one strand of the RNAi agent comprises a 5′ overhang of at least 1 nucleotide. In certain embodiments, at least one strand comprises a 5′ overhang of at least 2 nucleotides, e.g., 2, 3, 4, 5, 6, 7, 9, 10, 11, 12, 13, 14, or 15 nucleotides. In still other embodiments, both the 3′ and the 5′ end of one strand of the RNAi agent comprise an overhang of at least 1 nucleotide. 
     In one embodiment, the antisense strand of a dsRNA has a 1-10 nucleotide, e.g., a 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 nucleotide, overhang at the 3′-end and/or the 5′-end. In one embodiment, the sense strand of a dsRNA has a 1-10 nucleotide, e.g., a 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 nucleotide, overhang at the 3′-end and/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, 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 another embodiment, one or more of the nucleotides in the overhang is replaced with a nucleoside thiophosphate. 
     The terms “blunt” or “blunt ended” as used herein in reference to a dsRNA mean that there are no unpaired nucleotides or nucleotide analogs at a given terminal end of a dsRNA, i.e., no nucleotide overhang. One or both ends of a dsRNA can be blunt. Where both ends of a dsRNA are blunt, the dsRNA is said to be blunt ended. To be clear, a “blunt ended” dsRNA is a dsRNA that is blunt at both ends, i.e., no nucleotide overhang at either end of the molecule. Most often such a molecule will be double stranded over its entire length. 
     The term “antisense strand” or “guide strand” refers to the strand of an iRNA, e.g., a dsRNA, which includes a region that is substantially complementary to a target sequence, e.g., a GCK 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 GCK nucleotide sequence, as defined herein. Where the region of complementarity is not fully complementary to the target sequence, the mismatches can be in the internal or terminal regions of the molecule. Generally, the most tolerated mismatches are in the terminal regions, e.g., within 5, 4, 3, or 2 nucleotides of the 5′- and/or 3′-terminus 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, 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 and/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 an iRNA 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 GCK). For example, a polynucleotide is complementary to at least a part of a GCK mRNA if the sequence is substantially complementary to a non-interrupted portion of an mRNA encoding GCK. 
     Accordingly, in some embodiments, the antisense strand polynucleotides disclosed herein are fully complementary to the target GCK sequence. In other embodiments, the antisense strand polynucleotides disclosed herein are substantially complementary to the target GCK sequence and comprise a contiguous nucleotide sequence which is at least about 80% complementary over its entire length to the equivalent region of the nucleotide sequence of SEQ ID NO:1, or a fragment of SEQ ID NO:1, such as about 85%, about 86%, about 87%, about 88%, about 89%, about 90%, about % 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, or about 99% complementary. 
     In other embodiments, the antisense polynucleotides disclosed herein are substantially complementary to the target GCK sequence and comprise a contiguous nucleotide sequence which is at least about 80% complementary over its entire length to any one of the sense strand nucleotide sequences in any one of Tables 2, 3, 6, and 7, or a fragment of any one of the sense strand nucleotide sequences in any one of Tables 2, 3, 6, and 7, such as at least 85%, 90%, 95% complementary, or 100% complementary. 
     In one embodiment, an RNAi agent of the invention includes a sense strand that is substantially complementary to an antisense polynucleotide which, in turn, is complementary to a target GCK sequence, and wherein the sense strand polynucleotide comprises a contiguous nucleotide sequence which is at least about 80% complementary over its entire length to the equivalent region of the nucleotide sequence of SEQ ID NO:5, or a fragment of any one of SEQ ID NO:5, such as about 85%, about 86%, about 87%, about 88%, about 89%, about 90%, about % 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, or about 99% complementary. 
     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 GCK,” as used herein, includes inhibition of expression of any GCK gene (such as, e.g., a mouse GCK gene, a rat GCK gene, a monkey GCK gene, or a human GCK gene) as well as variants or mutants of a GCK gene that encode a GCK protein. 
     “Inhibiting expression of a GCK gene” includes any level of inhibition of a GCK gene, e.g., at least partial suppression of the expression of a GCK gene, such as an inhibition by at least about 20%. In certain embodiments, inhibition is by at least about 25%, at least about 30%, at least about 35%, at least about 40%, at least about 45%, at least about 50%, at least about 55%, at least about 60%, at least about 65%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99%. 
     The expression of a GCK gene may be assessed based on the level of any variable associated with GCK gene expression, e.g., GCK mRNA level or GCK protein level. The expression of a GCK may also be assessed indirectly based on, e.g., blood glucose levels, serum ketone body levels, and/or serum fatty acid levels. Inhibition may be assessed by a change in an absolute or relative level of one or more of these variables compared with a control level. The control level may be any type of control level that is utilized in the art, e.g., a pre-dose baseline level, or a level determined from a similar subject, cell, or sample that is untreated or treated with a control (such as, e.g., buffer only control or inactive agent control). 
     In one embodiment, at least partial suppression of the expression of a GCK gene, is assessed by a reduction of the amount of GCK mRNA which can be isolated from or detected in a first cell or group of cells in which a GCK gene is transcribed and which has or have been treated such that the expression of a GCK gene is inhibited, as compared to a second cell or group of cells substantially identical to the first cell or group of cells but which has or have not been so treated (control cells). The degree of inhibition may be expressed in terms of: 
     
       
         
           
             
               
                 
                   
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     The phrase “contacting a cell with an RNAi agent,” such as a dsRNA, as used herein, includes contacting a cell by any possible means. Contacting a cell with an RNAi agent includes contacting a cell in vitro with the iRNA or contacting a cell in vivo with the iRNA. The contacting may be done directly or indirectly. Thus, for example, the RNAi agent may be put into physical contact with the cell by the individual performing the method, or alternatively, the RNAi agent may be put into a situation that will permit or cause it to subsequently come into contact with the cell. 
     Contacting a cell in vitro may be done, for example, by incubating the cell with the RNAi agent. Contacting a cell in vivo may be done, for example, by injecting the RNAi agent into or near the tissue where the cell is located, or by injecting the RNAi agent 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 RNAi agent may contain and/or be coupled to a ligand, e.g., GalNAc3, that directs the RNAi agent 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 RNAi agent and subsequently transplanted into a subject. 
     In one embodiment, 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 diffusive or active cellular processes, or by auxiliary agents or devices. Introducing an iRNA into a cell may be in vitro and/or in vivo. For example, for in vivo introduction, iRNA can be injected into a tissue site or administered systemically. In vivo delivery can also be done by a beta-glucan delivery system, such as those described in U.S. Pat. Nos. 5,032,401 and 5,607,677, and U.S. Publication No. 2005/0281781, the entire contents of which are hereby incorporated herein by reference. In vitro introduction into a cell includes methods known in the art such as electroporation and lipofection. Further approaches are described herein below and/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 camel, a llama, a horse, a goat, a rabbit, a sheep, a hamster, a guinea pig, a cat, a dog, a rat, a mouse, a horse, and a whale), or a bird (e.g., a duck or a goose). It is understood that the sequence of the GCK gene must be sufficiently complementary to the antisense strand of the iRNA agent for the agent to be used in the indicated species. 
     In an embodiment, the subject is a human, such as a human being treated or assessed for a disease, disorder or condition that would benefit from reduction in GCK expression; a human at risk for a disease, disorder or condition that would benefit from reduction in GCK expression; a human having a disease, disorder or condition that would benefit from reduction in GCK expression; and/or human being treated for a disease, disorder or condition that would benefit from reduction in GCK expression as described herein. 
     As used herein, the terms “treating” or “treatment” refer to a beneficial or desired result including, such as increasing blood glucose levels in a subject. The terms “treating” or “treatment” also include, but are not limited to, alleviation or amelioration of one or more symptoms of a glycogen storage disease (GSD), e.g., type Ia GSD, or at least one sign or symptom associated with GSD, such as hypoglycemia, lactic acidosis, hyperuricemia, hyperlipidemia, hepatomegaly, kidney disease as a result of glycogen accumulation, hunger, jitteriness, lethargy, apnea, seizures, diaphoresis, confusion, headaches, dizziness, unusual mood or behavior changes, loss of consciousness, coma, muscle cramps, bleeding diathesis, short stauture, osteoporosis, delayed puberty, gout, renal disease, systemic hypertension, pulmonary hypertension, hepatic adenomas, pancreatitis, anemia, vitamin D deficiency, polycystic ovaries, irregular menstrual cycles, menorrhagia, and eruptive xanthomata. 
     “Treatment” can also mean prolonging survival as compared to expected survival in the absence of treatment. 
     By “lower” in the context of a disease marker or symptom is meant a statistically significant decrease in such level. The decrease can be, for example, at least 10%, at least 20%, at least 30%, at least 40%, or more, down to a level accepted as within the range of normal for an individual without such disorder, or to below the level of detection of the assay. In certain embodiments, the decrease is down to a level accepted as within the range of normal for an individual without such disorder which can also be referred to as a normalization of a level. For example, lowering cholesterol to 180 mg/dl or lower would be considered to be within the range of normal for a subject. A subject having a cholesterol level of 230 mg/dl with a cholesterol level decreased to 210 mg/dl would have a cholesterol level that was decreased by 40% towards normal (230−210/230−180=20/50=40% reduction). In certain embodiments, the reduction is the normalization of the level of a sign or symptom of a disease, a reduction in the difference between the subject level of a sign of the disease and the normal level of the sign for the disease (e.g., to the upper level of normal when the value for the subject must be decreased to reach a normal value, and to the lower level of normal when the value for the subject must be increased to reach a normal level). In certain embodiments, the methods include a clinically relevant inhibition of expression of a GCK gene, e.g. as demonstrated by a clinically relevant outcome after treatment of a subject with an agent to reduce the expression of a GCK gene. 
     As used herein, “prevention” or “preventing,” when used in reference to a disease, disorder or condition thereof, that would benefit from a reduction in expression of a GCK gene, refers to a reduction in the likelihood that a subject will develop a symptom associated with such disease, disorder, or condition, e.g., a glycogen storage disease (GSD), e.g., type Ia GSD. The likelihood of developing type Ia GSD is reduced, for example, when an individual having one or more risk factors for type Ia GSD, e.g., a genetic disorder, either fails to develop type Ia GSD, or signs or symptoms thereof, or develops type Ia GSD, or signs or symptoms thereof, with less severity relative to a population having the same risk factors and not receiving treatment as described herein. 
     The failure to develop a disease, disorder or condition, or the reduction in the development of a symptom associated with such a disease, disorder or condition (e.g., by at least about 10% on a clinically accepted scale for that disease or disorder), or the exhibition of delayed symptoms delayed (e.g., by days, weeks, months, or years) is considered effective prevention. Prevention can require administration of more than one dose of an agent described herein. 
     As used herein, the term “blood glucose level” refers to the level of glucose present in blood as determined by any routine method known in the art. It is understood that the glucose level in a subject sample is dependent on, for example, whether the subject has had a meal, whether the subject has fasted, the time of day and the level of activity of the subject. Therefore, the glucose level must be compared to an appropriate control to determine if the glucose level is, in fact, altered from a normal level or from a level obtained from the subject at an earlier time point, e.g., prior to treatment. In general, blood glucose levels, e.g., fasting blood glucose levels, in a subject that does not have a disease or disorder that would benefit from reduction in the levels of GCK as described herein is about 70 to about 120 mg/dL. 
     As used herein, the term “plasma lactate level” refers to the level of lactate present in blood as determined by any routine method known in the art. It is understood that the lactate level in a subject sample is dependent on, for example, whether the subject has fasted, the time of day and the level of activity of the subject. Therefore, the lactate level must be compared to an appropriate control to determine if the lactate level is, in fact, altered from a normal level or from a level obtained from the subject at an earlier time point, e.g., prior to treatment. In general, plasma lactate levels in a subject that does not have a disease or disorder that would benefit from reduction in the levels of GCK as described herein is about 0.5 to about 2.2 mmol/L. 
     As used herein, the term “plasma uric acid level” refers to the level of uric acid present in blood as determined by any routine method known in the art. It is understood that the lactate level in a subject sample is dependent on, for example, whether the subject has fasted, the time of day and the level of activity of the subject. Therefore, the uric acid level must be compared to an appropriate control to determine if the uric acid level is, in fact, altered from a normal level or from a level obtained from the subject at an earlier time point, e.g., prior to treatment. In general, plasma uric acid levels in a subject that does not have a disease or disorder that would benefit from reduction in the levels of GCK as described herein is about 2.0 to about 5.0 mg/dL. 
     As used herein, the term “plasma triglyceride level” refers to the level of triglycerides present in blood as determined by any routine method known in the art. It is understood that the triglyceride level in a subject sample is dependent on, for example, whether the subject has fasted, the time of day and the level of activity of the subject. Therefore, the triglyceride level must be compared to an appropriate control to determine if the triglyceride level is, in fact, altered from a normal level or from a level obtained from the subject at an earlier time point, e.g., prior to treatment. In general, plasma triglyceride levels in a subject that does not have a disease or disorder that would benefit from reduction in the levels of GCK as described herein is about 150 to about 200 mg/dL. 
     As used herein, the term “total cholesterol level” refers to the level of high density lipoprotein (HDL) plus low density lipoprotein (LDL) plus 20% of the triglyceride level as determined by any routine method known in the art. It is understood that the total cholesterol level in a subject sample is dependent on, for example, whether the subject has fasted, the time of day and the level of activity of the subject. Therefore, the total cholesterol level must be compared to an appropriate control to determine if the total cholesterol level is, in fact, altered from a normal level or from a level obtained from the subject at an earlier time point, e.g., prior to treatment. In general, plasma cholesterol levels in a subject that does not have a disease or disorder that would benefit from reduction in the levels of GCK as described herein is about 100 to about 200 mg/dL. 
     As used herein, the term “hepatomegaly” refers to swelling of the liver beyond its normal size as determined by any routine method known in the art. It is understood that the size and weight of the liver in a subject that does not have a disease or disorder that would benefit from reduction in the levels of GCK as described herein increases with age and body weight. Sex and body shape also influence the size of the liver, e.g., by percussion, the mean liver size is about 7.5 centimeters in adult women and about 10.5 centimeters in adult men; it may be about 3 centimeters larger or smaller and still be normal. 
     As used herein, a “disease or disorder that would benefit from reduction in GCK expression” is a disease or disorder associated with or caused by a clinically relevant hypoglycemia. For example, this term includes any disorder, disease or condition resulting in one or more signs or symptoms of a glycogen storage disease (GSD), e.g., type Ia GSD, including, but not limited to, hypoglycemia, lactic acidosis, hyperuricemia, hyperlipidemia, hepatomegaly, kidney disease as a result of glycogen accumulation, hunger, jitteriness, lethargy, apnea, seizures, diaphoresis, confusion, headaches, dizziness, unusual mood or behavior changes, loss of consciousness, coma, muscle cramps, bleeding diathesis, short stauture, osteoporosis, delayed puberty, gout, renal disease, systemic hypertension, pulmonary hypertension, hepatic adenomas, pancreatitis, anemia, vitamin D deficiency, polycystic ovaries, irregular menstrual cycles, menorrhagia, and eruptive xanthomata. In certain embodiments, a “disease or disorder that would benefit from reduction in GCK expression” meets the diagnostic requirements of a type Ia GSD. 
     “Glycogen storage disease” or “GSD” is an inherited genetic disorder due to an absence or deficiency of one of the enzymes responsible for making or breaking down glycogen in the body. This enzyme deficiency causes either abnormal tissue concentrations of glycogen or incorrectly or abnormally formed glycogen. There are about 11 known types of GSD, which are classified based on the missing or defective enzymes. For example, Type Ia GSD, the most common form of GSD, is caused by a genetic defect in the enzyme glucose-6-phosphatase. Hepatomegaly due to inappropriate glycogen accumulation and various metabolic disarrangements from inappropriate glucose-6-phosphate metabolism are predominant features of many of the various glycogen storage diseases, such as Types Ia, Ib, III, IV, VI and IX. Symptoms of GSD vary based on the enzyme that is missing and usually result from the buildup of glycogen or from the inability to produce glucose when needed. Glycogen storage disease is usually diagnosed in infancy or childhood, e.g., at about age 3-4. Affected children typically present with hepatomegaly, lactic acidosis, hyperuricemia, hyperlipidemia, hypertriglyceridemia and/or hypoglycemic seizures. Further, affected children often have doll-like faces with fat cheeks, relatively thin extremities, short stature, and protuberant abdomen. Xanthoma and diarrhea may be present. Impaired platelet function can lead to a bleeding tendency with frequent epistaxis. The diagnostic criteria for GSD, e.g., type Ia GSD, include, for example, fasting blood glucose concentration lower than 60 mg/dL; plasma lactate higher than 2.5 mmol/L; plasma uric acid higher than 5.0 mg/dL; triglycerides higher than 250 mg/dL; total plasma cholesterol higher than 200 mg/dL; administration of glucagon or epinephrine (i.e., a glucagon or epinephrine challenge test) causes little or no increase in blood glucose concentration, but both increase serum lactate concentrations significantly; histopathologic liver findings which include distention of the liver cells by glycogen and fat; PAS positive and diastase sensitive glycogen that is uniformly distributed within the cytoplasm; normal or only modestly increased glycogen; and large and numerous lipid vacuoles; biallelic mutations in either G6PC (GSDIa) or SLC37A4 (GSDIb) (Veiga-da-Cunha et al (1998) Am J Hum Genet.  63:976-83; Chou et al (2002)  Hum Mutat.  29:921-30; Matern et al (2002)  Eur J Pediatr.  161 Suppl 1:S10-9; Rake et al (2002)  Eur J Pediatr.  161 Suppl 1:S20-34; and Ekstein et al (2004) Am J Med Genet.  129A:162-4); deficient, e.g., (lower than 10% of normal) glucose-6-phosphatase (G6Pase) catalytic activity (the normal G6Pase enzyme activity level in liver is 3.50±0.8 μmol/min/g tissue, although in rare individuals with milder clinical manifestations, the G6Pase enzyme activity can be higher, e.g., &gt;1.0 μmol/min/g tissue and &lt;2.0 μmol/min/g tissue. 
     “Therapeutically effective amount,” as used herein, is intended to include the amount of an RNAi agent that, when administered to a subject having a glycogen storage disease (GSD), e.g., type Ia GSD, is sufficient to effect treatment of the disease (e.g., by diminishing, ameliorating or maintaining the existing disease or one or more symptoms of disease). The “therapeutically effective amount” may vary depending on the RNAi agent, how the agent is administered, the disease and its severity and the history, age, weight, family history, genetic makeup, the types of preceding or concomitant treatments, if any, and other individual characteristics of the subject to be treated. 
     “Prophylactically effective amount,” as used herein, is intended to include the amount of an iRNA that, when administered to a subject having a glycogen storage disease (GSD), e.g., type Ia GSD, is sufficient to prevent or ameliorate the disease or one or more symptoms of the disease. Ameliorating the disease includes slowing the course of the disease or reducing the severity of later-developing disease. The “prophylactically effective amount” may vary depending on the iRNA, how the agent is administered, the degree of risk of disease, and the history, age, weight, family history, genetic makeup, the types of preceding or concomitant treatments, if any, and other individual characteristics of the patient to be treated. 
     A “therapeutically-effective amount” or “prophylacticaly effective amount” also includes an amount of an RNAi agent that produces some desired local or systemic effect at a reasonable benefit/risk ratio applicable to any treatment. 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, and/or dosage forms which are, within the scope of sound medical judgment, suitable for use in contact with the tissues of human subjects and animal subjects without excessive toxicity, irritation, allergic response, or other problem or complication, commensurate with a reasonable benefit/risk ratio. 
     The phrase “pharmaceutically-acceptable carrier” as used herein means a pharmaceutically-acceptable material, composition or vehicle, such as a liquid or solid filler, diluent, excipient, manufacturing aid (e.g., lubricant, talc magnesium, calcium or zinc stearate, or steric acid), or solvent encapsulating material, involved in carrying or transporting the subject compound from one organ, or portion of the body, to another organ, or portion of the body. Each carrier must be “acceptable” in the sense of being compatible with the other ingredients of the formulation and not injurious to the subject being treated. Some examples of materials which can serve as pharmaceutically-acceptable carriers include: (1) sugars, such as lactose, glucose and sucrose; (2) starches, such as corn starch and potato starch; (3) cellulose, and its derivatives, such as sodium carboxymethyl cellulose, ethyl cellulose and cellulose acetate; (4) powdered tragacanth; (5) malt; (6) gelatin; (7) lubricating agents, such as magnesium state, sodium lauryl sulfate and talc; (8) excipients, such as cocoa butter and suppository waxes; (9) oils, such as peanut oil, cottonseed oil, safflower oil, sesame oil, olive oil, corn oil and soybean oil; (10) glycols, such as propylene glycol; (11) polyols, such as glycerin, sorbitol, mannitol and polyethylene glycol; (12) esters, such as ethyl oleate and ethyl laurate; (13) agar; (14) buffering agents, such as magnesium hydroxide and aluminum hydroxide; (15) alginic acid; (16) pyrogen-free water; (17) isotonic saline; (18) Ringer&#39;s solution; (19) ethyl alcohol; (20) pH buffered solutions; (21) polyesters, polycarbonates and/or polyanhydrides; (22) bulking agents, such as polypeptides and amino acids (23) serum component, such as serum albumin, HDL and LDL; and (22) other non-toxic compatible substances employed in pharmaceutical formulations. 
     The term “sample,” as used herein, includes a collection of similar fluids, cells, or tissues isolated from a subject, as well as fluids, cells, or tissues present within a subject. Examples of biological fluids include blood, serum and serosal fluids, plasma, 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 some embodiments, a “sample derived from a subject” refers to blood drawn from the subject or plasma isolated therefrom, saliva, or urine, typically a 24 hour urine sample. 
     II. iRNAs of the Invention 
     Described herein are iRNAs which inhibit the expression of a GCK gene. In one embodiment, the iRNA agent includes double stranded ribonucleic acid (dsRNA) molecules for inhibiting the expression of a GCK gene in a cell, such as a cell within a subject, e.g., a mammal, such as a human having a glycogen storage disease (GSD), e.g., type Ia GSD. The dsRNA 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 GCK gene. The region of complementarity is about 30 nucleotides or less in length (e.g., about 30, 29, 28, 27, 26, 25, 24, 23, 22, 21, 20, 19, or 18 nucleotides or less in length). Upon contact with a cell expressing the GCK gene, the iRNA inhibits the expression of the GCK gene (e.g., a human, a primate, a non-primate, or a bird GCK gene) by at least about 10% as assayed by, for example, a PCR or branched DNA (bDNA)-based method, or by a protein-based method, such as by immunofluorescence analysis, using, for example, western blotting, or flowcytometric techniques. In preferred embodiments, inhibiton of expression is deteremined by the qPCR method provided in the examples. For in vitro assessment of activity, percent inhibition is determined using the methods provided herein at a single dose at, for example, a 10 nM duplex final concentration. For in vivo studies, the level after treatment can be compared to, for example, an appropriate historical control or a pooled population sample control to determine the level of reduction, e.g., when a baseline value is no available for the subject. 
     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 GCK 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 between 15 and 30 base pairs in length, e.g., between, 15-29, 15-28, 15-27, 15-26, 15-25, 15-24, 15-23, 15-22, 15-21, 15-20, 15-19, 15-18, 15-17, 18-30, 18-29, 18-28, 18-27, 18-26, 18-25, 18-24, 18-23, 18-22, 18-21, 18-20, 19-30, 19-29, 19-28, 19-27, 19-26, 19-25, 19-24, 19-23, 19-22, 19-21, 19-20, 20-30, 20-29, 20-28, 20-27, 20-26, 20-25, 20-24, 20-23, 20-22, 20-21, 21-30, 21-29, 21-28, 21-27, 21-26, 21-25, 21-24, 21-23, or 21-22 base pairs in length. Ranges and lengths intermediate to the above recited ranges and lengths are also contemplated to be part of the invention. 
     Similarly, the region of complementarity to the target sequence is between 15 and 30 nucleotides in length, e.g., between 15-29, 15-28, 15-27, 15-26, 15-25, 15-24, 15-23, 15-22, 15-21, 15-20, 15-19, 15-18, 15-17, 18-30, 18-29, 18-28, 18-27, 18-26, 18-25, 18-24, 18-23, 18-22, 18-21, 18-20, 19-30, 19-29, 19-28, 19-27, 19-26, 19-25, 19-24, 19-23, 19-22, 19-21, 19-20, 20-30, 20-29, 20-28, 20-27, 20-26, 20-25, 20-24, 20-23, 20-22, 20-21, 21-30, 21-29, 21-28, 21-27, 21-26, 21-25, 21-24, 21-23, or 21-22 nucleotides in length. Ranges and lengths intermediate to the above recited ranges and lengths are also contemplated to be part of the invention. 
     In some embodiments, the dsRNA is between about 15 and about 23 nucleotides in length, or between about 25 and 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 can serve as substrates for Dicer. As the ordinarily skilled person will also recognize, the region of an RNA targeted for cleavage will most often be part of a larger RNA molecule, often an mRNA molecule. Where relevant, a “part” of an mRNA target is a contiguous sequence of an mRNA target of sufficient length to allow it to be a substrate for RNAi-directed cleavage (i.e., cleavage through a RISC pathway). 
     One of skill in the art will also recognize that the duplex region is a primary functional portion of a dsRNA, e.g., a duplex region of about 9 to 36 base pairs, e.g., about 10-36, 11-36, 12-36, 13-36, 14-36, 15-36, 9-35, 10-35, 11-35, 12-35, 13-35, 14-35, 15-35, 9-34, 10-34, 11-34, 12-34, 13-34, 14-34, 15-34, 9-33, 10-33, 11-33, 12-33, 13-33, 14-33, 15-33, 9-32, 10-32, 11-32, 12-32, 13-32, 14-32, 15-32, 9-31, 10-31, 11-31, 12-31, 13-32, 14-31, 15-31, 15-30, 15-29, 15-28, 15-27, 15-26, 15-25, 15-24, 15-23, 15-22, 15-21, 15-20, 15-19, 15-18, 15-17, 18-30, 18-29, 18-28, 18-27, 18-26, 18-25, 18-24, 18-23, 18-22, 18-21, 18-20, 19-30, 19-29, 19-28, 19-27, 19-26, 19-25, 19-24, 19-23, 19-22, 19-21, 19-20, 20-30, 20-29, 20-28, 20-27, 20-26, 20-25, 20-24, 20-23, 20-22, 20-21, 21-30, 21-29, 21-28, 21-27, 21-26, 21-25, 21-24, 21-23, or 21-22 base pairs. 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 GCK expression is not generated in the target cell by cleavage of a larger dsRNA. 
     A dsRNA as described herein can further include one or more single-stranded nucleotide overhangs e.g., 1, 2, 3, or 4 nucleotides. dsRNAs having at least one nucleotide overhang can have unexpectedly 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 either an antisense or sense strand of a dsRNA. In certain embodiments, longer, extended overhangs are possible. 
     A dsRNA can be synthesized by standard methods known in the art as further discussed below, e.g., by use of an automated DNA synthesizer, such as are commercially available from, for example, Biosearch™, Applied Biosystems™, Inc. 
     iRNA compounds of the invention may be prepared using a two-step procedure. First, the individual strands of the double stranded RNA molecule are prepared separately. Then, the component strands are annealed. The individual strands of the siRNA compound can be prepared using solution-phase or solid-phase organic synthesis or both. Organic synthesis offers the advantage that the oligonucleotide strands comprising unnatural or modified nucleotides can be easily prepared. Single-stranded oligonucleotides of the invention can be prepared using solution-phase or solid-phase organic synthesis or both. 
     In certain aspects, a dsRNA of the invention includes at least two nucleotide sequences, a sense sequence and an anti-sense sequence. The sense strand sequence is selected from the group of sequences provided in any of Tables 2, 3, 6, and 7 and the corresponding nucleotide sequence of the antisense strand of the sense strand is selected from the group of sequences provided in any of Tables 2, 3, 6, and 7. 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 GCK gene. As such, in this aspect, a dsRNA will include two oligonucleotides, where one oligonucleotide is described as the sense strand in any of Tables 2, 3, 6, and 7, and the second oligonucleotide is described as the corresponding antisense strand of the sense strand in any of Tables 2, 3, 6, and 7. In one embodiment, the substantially complementary sequences of the dsRNA are contained on separate oligonucleotides. In another embodiment, the substantially complementary sequences of the dsRNA are contained on a single oligonucleotide. 
     It will be understood that, although the sequences in Tables 3 and 7 are described as modified and/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 any one of Tables 3 and 7 that is un-modified, un-conjugated, and/or modified and/or conjugated differently than described therein. 
     In another aspect, a double stranded ribonucleic acid (dsRNA) of the invention for inhibiting expression of GCK comprises, consists essentially of, or consists of a sense strand and an antisense strand, wherein the sense strand comprises the nucleotide sequence of a sense strand in any of Tables 2, 3, 6, and 7 and the antisense strand comprises the nucleotide sequence of the corresponding antisense strand in any of Tables 2, 3, 6, and 7. 
     The skilled person is well aware that dsRNAs having a duplex structure of between about 20 and 23 base pairs, e.g., 21, base pairs have been hailed as particularly effective in inducing RNA interference (Elbashir et al., (2001)  EMBO J.,  20:6877-6888). However, others have found that shorter or longer RNA duplex structures can also be effective (Chu and Rana (2007) RNA 14:1714-1719; Kim et al. (2005)  Nat Biotech  23:222-226). In the embodiments described above, by virtue of the nature of the oligonucleotide sequences provided herein, dsRNAs described herein can include at least one strand of a length of minimally 21 nucleotides. It can be reasonably expected that shorter duplexes minus only a few nucleotides on one or both ends can be similarly effective as compared to the dsRNAs described above. Hence, dsRNAs having a sequence of at least 15, 16, 17, 18, 19, 20, or more contiguous nucleotides derived from one of the sequences provided herein, and differing in their ability to inhibit the expression of a GCK 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 described herein identify a site(s) in a GCK transcript that is susceptible to RISC-mediated cleavage. As such, the present invention further features iRNAs that target within this site(s). 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 15 contiguous nucleotides from one of the sequences provided herein coupled to additional nucleotide sequences taken from the region contiguous to the selected sequence in a GCK gene. 
     While a target sequence is generally about 15-30 nucleotides in length, there is wide variation in the suitability of particular sequences in this range for directing cleavage of any given target RNA. Various software packages and the guidelines set out herein provide guidance for the identification of optimal target sequences for any given gene target, but an empirical approach can also be taken in which a “window” or “mask” of a given size (as a non-limiting example, 21 nucleotides) is literally or figuratively (including, e.g., in silico) placed on the target RNA sequence to identify sequences in the size range that can serve as target sequences. By moving the sequence “window” progressively one nucleotide upstream or downstream of an initial target sequence location, the next potential target sequence can be identified, until the complete set of possible sequences is identified for any given target size selected. This process, coupled with systematic synthesis and testing of the identified sequences (using assays as described herein or as known in the art) to identify those sequences that perform optimally can identify those RNA sequences that, when targeted with an iRNA agent, mediate the best inhibition of target gene expression. Thus, while the sequences identified herein represent effective target sequences, it is contemplated that further optimization of inhibition efficiency can be achieved by progressively “walking the window” one nucleotide upstream or downstream of the given sequences to identify sequences with equal or better inhibition characteristics. 
     Further, it is contemplated that for any sequence identified herein, further optimization could be achieved by systematically either adding or removing nucleotides to generate longer or shorter sequences and testing those sequences generated by walking a window of the longer or shorter size up or down the target RNA from that point. Again, coupling this approach to generating new candidate targets with testing for effectiveness of iRNAs based on those target sequences in an inhibition assay as known in the art and/or as described herein can lead to further improvements in the efficiency of inhibition. Further still, such optimized sequences can be adjusted by, e.g., the introduction of modified nucleotides as described herein or as known in the art, addition or changes in overhang, or other modifications as known in the art and/or discussed herein to further optimize the molecule (e.g., increasing serum stability or circulating half-life, increasing thermal stability, enhancing transmembrane delivery, targeting to a particular location or cell type, increasing interaction with silencing pathway enzymes, increasing release from endosomes) as an expression inhibitor. 
     An iRNA agent as described herein can contain one or more mismatches to the target sequence. In one embodiment, an iRNA as described herein contains no more than 3 mismatches. If the antisense strand of the iRNA contains mismatches to a target sequence, it is preferable that the area of mismatch is not located in the center of the region of complementarity. If the antisense strand of the iRNA contains mismatches to the target sequence, it is preferable that the mismatch be restricted to be within the last 5 nucleotides from either the 5′- or 3′-end of the region of complementarity. For example, for a 23 nucleotide iRNA agent the strand which is complementary to a region of a GCK gene, generally does not contain any mismatch within the central 13 nucleotides. The methods described herein or methods known in the art can be used to determine whether an iRNA containing a mismatch to a target sequence is effective in inhibiting the expression of a GCK gene. Consideration of the efficacy of iRNAs with mismatches in inhibiting expression of a GCK gene is important, especially if the particular region of complementarity in a GCK gene is known to have polymorphic sequence variation within the population. 
     III. Modified iRNAs of the Invention 
     In one embodiment, the RNA of the iRNA of the invention e.g., a dsRNA, is un-modified, and does not comprise, e.g., chemical modifications and/or conjugations known in the art and described herein. In another embodiment, 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 of the invention are modified. iRNAs of the invention in which “substantially all of the nucleotides are modified” are largely but not wholly modified and can include not more than 5, 4, 3, 2, or 1 unmodified nucleotides. 
     The nucleic acids featured in the invention can be synthesized and/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; and/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. RE39464, the entire contents of each of which are hereby incorporated herein by reference. 
     Modified RNA backbones that do not include a phosphorus atom therein have backbones that are formed by short chain alkyl or cycloalkyl internucleoside linkages, mixed heteroatoms and alkyl or cycloalkyl internucleoside linkages, or one or more short chain heteroatomic or heterocyclic internucleoside linkages. These include those having morpholino linkages (formed in part from the sugar portion of a nucleoside); siloxane backbones; sulfide, sulfoxide and sulfone backbones; formacetyl and thioformacetyl backbones; methylene formacetyl and thioformacetyl backbones; alkene containing backbones; sulfamate backbones; methyleneimino and methylenehydrazino backbones; sulfonate and sulfonamide backbones; amide backbones; and others having mixed N, O, S, and CH 2  component parts. 
     Representative U.S. patents that teach the preparation of the above oligonucleosides include, but are not limited to, U.S. Pat. Nos. 5,034,506; 5,166,315; 5,185,444; 5,214,134; 5,216,141; 5,235,033; 5,64,562; 5,264,564; 5,405,938; 5,434,257; 5,466,677; 5,470,967; 5,489,677; 5,541,307; 5,561,225; 5,596,086; 5,602,240; 5,608,046; 5,610,289; 5,618,704; 5,623,070; 5,663,312; 5,633,360; 5,677,437; and, 5,677,439, the entire contents of each of which are hereby incorporated herein by reference. 
     In other embodiments, suitable RNA mimetics are contemplated for use in iRNAs, in which both the sugar and the internucleoside linkage, i.e., the backbone, of the nucleotide units are replaced with novel groups. The base units are maintained for hybridization with an appropriate nucleic acid target compound. One such oligomeric compound, an RNA mimetic that has been shown to have excellent hybridization properties, is referred to as a peptide nucleic acid (PNA). In PNA compounds, the sugar backbone of an RNA is replaced with an amide containing backbone, in particular an aminoethylglycine backbone. The nucleobases are retained and are bound directly or indirectly to aza nitrogen atoms of the amide portion of the backbone. Representative U.S. patents that teach the preparation of PNA compounds include, but are not limited to, U.S. Pat. Nos. 5,539,082; 5,714,331; and 5,719,262, the entire contents of each of which are hereby incorporated herein by reference. Additional PNA compounds suitable for use in the 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 -4 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 U.S. patents that teach the preparation of such modified sugar structures include, but are not limited to, U.S. Pat. Nos. 4,981,957; 5,118,800; 5,319,080; 5,359,044; 5,393,878; 5,446,137; 5,466,786; 5,514,785; 5,519,134; 5,567,811; 5,576,427; 5,591,722; 5,597,909; 5,610,300; 5,627,053; 5,639,873; 5,646,265; 5,658,873; 5,670,633; and 5,700,920, certain of which are commonly owned with the instant application. The entire contents of each of the foregoing are hereby incorporated herein by reference. 
     An 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 5-methylcytosine (5-me-C), 5-hydroxymethyl cytosine, xanthine, hypoxanthine, 2-aminoadenine, 6-methyl and other alkyl derivatives of adenine and guanine, 2-propyl and other alkyl derivatives of adenine and guanine, 2-thiouracil, 2-thiothymine and 2-thiocytosine, 5-halouracil and cytosine, 5-propynyl uracil and cytosine, 6-azo uracil, cytosine and thymine, 5-uracil (pseudouracil), 4-thiouracil, 8-halo, 8-amino, 8-thiol, 8-thioalkyl, 8-hydroxyl anal other 8-substituted adenines and guanines, 5-halo, particularly 5-bromo, 5-trifluoromethyl and other 5-substituted uracils and cytosines, 7-methylguanine and 7-methyladenine, 8-azaguanine and 8-azaadenine, 7-deazaguanine and 7-daazaadenine and 3-deazaguanine and 3-deazaadenine. Further nucleobases include those disclosed in U.S. Pat. No. 3,687,808, those disclosed in Modified Nucleosides in Biochemistry, Biotechnology and Medicine, Herdewijn, P. ed. Wiley-VCH, 2008; those disclosed in The Concise Encyclopedia Of Polymer Science And Engineering, pages 858-859, Kroschwitz, J. L, ed. John Wiley &amp; Sons, 1990, these disclosed by Englisch et al., (1991)  Angewandte Chemie, International Edition,  30:613, and those disclosed by Sanghvi, Y S., Chapter 15, dsRNA Research and Applications, pages 289-302, Crooke, S. T. and Lebleu, B., Ed., CRC Press, 1993. Certain of these nucleobases are particularly useful for increasing the binding affinity of the oligomeric compounds featured in the invention. These include 5-substituted pyrimidines, 6-azapyrimidines and N-2, N-6 and 0-6 substituted purines, including 2-aminopropyladenine, 5-propynyluracil and 5-propynylcytosine. 5-methylcytosine substitutions have been shown to increase nucleic acid duplex stability by 0.6-1.2° C. (Sanghvi, Y. S., Crooke, S. T. and Lebleu, B., Eds., dsRNA Research and Applications, CRC Press, Boca Raton, 1993, pp. 276-278) and are exemplary base substitutions, even more particularly when combined with 2′-O-methoxyethyl sugar modifications. 
     Representative U.S. patents that teach the preparation of certain of the above noted modified nucleobases as well as other modified nucleobases include, but are not limited to, the above noted U.S. Pat. Nos. 3,687,808, 4,845,205; 5,130,30; 5,134,066; 5,175,273; 5,367,066; 5,432,272; 5,457,187; 5,459,255; 5,484,908; 5,502,177; 5,525,711; 5,552,540; 5,587,469; 5,594,121, 5,596,091; 5,614,617; 5,681,941; 5,750,692; 6,015,886; 6,147,200; 6,166,197; 6,222,025; 6,235,887; 6,380,368; 6,528,640; 6,639,062; 6,617,438; 7,045,610; 7,427,672; and 7,495,088, the entire contents of each of which are hereby incorporated herein by reference. 
     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 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 US Patent Publication Nos. 2013/0096289; 2013/0011922; and 2011/0313020, 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 bicyclic sugar moities. A “bicyclic sugar” is a furanosyl ring modified by the bridging of two atoms. A″bicyclic nucleoside” (“BNA”) is a nucleoside having a sugar moiety comprising a bridge connecting two carbon atoms of the sugar ring, thereby forming a bicyclic ring system. In certain embodiments, the bridge connects the 4′-carbon and the 2′-carbon of the sugar ring. Thus, in some embodiments an agent of the 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′-CH2-O-2′ bridge. This structure effectively “locks” the ribose in the 3′-endo structural conformation. The addition of locked nucleic acids to siRNAs has been shown to increase siRNA stability in serum, and to reduce off-target effects (Elmen, J. et al., (2005)  Nucleic Acids Research  33(1):439-447; Mook, O R. et al., (2007)  Mol Canc Ther  6(3):833-843; Grunweller, A. et al., (2003)  Nucleic Acids Research  31(12):3185-3193). Examples of bicyclic nucleosides for use in the polynucleotides of the 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′-(CH2)-O-2′ (LNA); 4′-(CH2)-S-2′; 4′-(CH2)2-O-2′ (ENA); 4′-CH(CH3)-O-2′ (also referred to as “constrained ethyl” or “cEt”) and 4′-CH(CH2OCH3)-O-2′ (and analogs thereof; see, e.g., U.S. Pat. No. 7,399,845); 4′-C(CH3)(CH3)-O-2′ (and analogs thereof; see e.g., U.S. Pat. No. 8,278,283); 4′-CH2-N(OCH3)-2′ (and analogs thereof; see e.g., U.S. Pat. No. 8,278,425); 4′-CH2-O—N(CH3)-2′ (see, e.g., U.S. Patent Publication No. 2004/0171570); 4′-CH2-N(R)—O-2′, wherein R is H, C1-C12 alkyl, or a protecting group (see, e.g., U.S. Pat. No. 7,427,672); 4′-CH2-C(H)(CH3)-2′ (see, e.g., Chattopadhyaya et al.,  J. Org. Chem.,  2009, 74, 118-134); and 4′-CH2C(═CH2)-2′ (and analogs thereof; see, e.g., U.S. Pat. No. 8,278,426). The entire contents of each of the foregoing are hereby incorporated herein by reference. 
     Additional representative U.S. Patents and US 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(CH3)-0-2′ 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, US Patent Publication No. 2013/0190383; and PCT publication WO 2013/036868, 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′-0-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 RNAi agent. Suitable phosphate mimics are disclosed in, for example US Patent Publication No. 2012/0157511, the entire contents of which are incorporated herein by reference. 
     IV. iRNAs Conjugated to Ligands 
     Another modification of the RNA of an iRNA of the invention involves chemically linking to the RNA one or more ligands, moieties or conjugates that enhance the activity, cellular distribution or cellular uptake of the iRNA. Such moieties include but are not limited to lipid moieties such as a cholesterol moiety (Letsinger et al., (1989)  Proc. Natl. Acid. Sci. USA,  86: 6553-6556), cholic acid (Manoharan et al., (1994)  Biorg. Med. Chem. Let.,  4:1053-1060), a thioether, e.g., beryl-S-tritylthiol (Manoharan et al., (1992)  Ann. N.Y. Acad. Sci.,  660:306-309; Manoharan et al., (1993)  Biorg. Med. Chem. Let.,  3:2765-2770), a thiocholesterol (Oberhauser et al., (1992)  Nucl. Acids Res.,  20:533-538), an aliphatic chain, e.g., dodecandiol or undecyl residues (Saison-Behmoaras et al., (1991)  EMBO J,  10:1111-1118; Kabanov et al., (1990)  FEBS Lett.,  259:327-330; Svinarchuk et al., (1993)  Biochimie,  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., (1995)  Tetrahedron Lett.,  36:3651-3654; Shea et al., (1990)  Nucl. Acids Res.,  18:3777-3783), a polyamine or a polyethylene glycol chain (Manoharan et al., (1995)  Nucleosides  &amp;  Nucleotides,  14:969-973), or adamantane acetic acid (Manoharan et al., (1995)  Tetrahedron Lett.,  36:3651-3654), a palmityl moiety (Mishra et al., (1995)  Biochim. Biophys. Acta,  1264:229-237), or an octadecylamine or hexylamino-carbonyloxycholesterol moiety (Crooke et al., (1996)  J. Pharmacol. Exp. Ther.,  277:923-937). 
     In one embodiment, 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 will not take part in duplex pairing in a duplexed nucleic acid. 
     Ligands can include a naturally occurring substance, such as a protein (e.g., human serum albumin (HSA), low-density lipoprotein (LDL), or globulin); carbohydrate (e.g., a dextran, pullulan, chitin, chitosan, inulin, cyclodextrin, 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, monovalent or multivalent galactose, N-acetyl-galactosamine, N-acetyl-gulucosamine multivalent mannose, multivalent fucose, glycosylated polyaminoacids, 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, ligands include monovalent or multivalent galactose. In certain embodiments, ligands include cholesterol. 
     Other examples of ligands include dyes, intercalating agents (e.g. acridines), cross-linkers (e.g. psoralene, mitomycin C), porphyrins (TPPC4, texaphyrin, Sapphyrin), polycyclic aromatic hydrocarbons (e.g., phenazine, dihydrophenazine), artificial endonucleases (e.g. EDTA), lipophilic molecules, e.g., cholesterol, cholic acid, adamantane acetic acid, 1-pyrene butyric acid, dihydrotestosterone, 1,3-Bis-O(hexadecyl)glycerol, geranyloxyhexyl group, hexadecylglycerol, borneol, menthol, 1,3-propanediol, heptadecyl group, palmitic acid, myristic acid, O3-(oleoyl)lithocholic acid, O3-(oleoyl)cholenic acid, dimethoxytrityl, or phenoxazine), and peptide conjugates (e.g., antennapedia peptide, Tat peptide), alkylating agents, phosphate, amino, mercapto, PEG (e.g., PEG-40K), MPEG, [MPEG]2, polyamino, alkyl, substituted alkyl, radiolabeled markers, enzymes, haptens (e.g. biotin), transport/absorption facilitators (e.g., aspirin, vitamin E, folic acid), synthetic ribonucleases (e.g., imidazole, bisimidazole, histamine, imidazole clusters, acridine-imidazole conjugates, Eu3+ complexes of tetraazamacrocycles), dinitrophenyl, HRP, or AP. 
     Ligands can be proteins, e.g., glycoproteins, or peptides, e.g., molecules having a specific affinity for a co-ligand, or antibodies e.g., an antibody, that binds to a specified cell type such as a 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-gulucosamine 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, and/or intermediate filaments. The drug can be, for example, taxon, vincristine, vinblastine, cytochalasin, nocodazole, japlakinolide, latrunculin A, phalloidin, swinholide A, indanocine, or myoservin. 
     In some embodiments, a ligand attached to an iRNA as described herein acts as a pharmacokinetic modulator (PK modulator). PK modulators include lipophiles, bile acids, steroids, phospholipid analogues, peptides, protein binding agents, PEG, vitamins etc. Exemplary PK modulators include, but are not limited to, cholesterol, fatty acids, cholic acid, lithocholic acid, dialkylglycerides, diacylglyceride, phospholipids, sphingolipids, naproxen, ibuprofen, vitamin E, biotin etc. Oligonucleotides that comprise a number of phosphorothioate linkages are also known to bind to serum protein, thus short oligonucleotides, e.g., oligonucleotides of about 5 bases, 10 bases, 15 bases or 20 bases, comprising multiple of phosphorothioate linkages in the backbone are also amenable to the present invention as ligands (e.g. as PK modulating ligands). In addition, aptamers that bind serum components (e.g. serum proteins) are also suitable for use as PK modulating ligands in the embodiments described herein. 
     Ligand-conjugated oligonucleotides of the invention may be synthesized by the use of an oligonucleotide that bears a pendant reactive functionality, such as that derived from the attachment of a linking molecule onto the oligonucleotide (described below). This reactive oligonucleotide may be reacted directly with commercially-available ligands, ligands that are synthesized bearing any of a variety of protecting groups, or ligands that have a linking moiety attached thereto. 
     The oligonucleotides used in the conjugates of the present invention may be conveniently and routinely made through the well-known technique of solid-phase synthesis. Equipment for such synthesis is sold by several vendors including, for example, Applied Biosystems™ (Foster City, Calif.). Any other means for such synthesis known in the art may additionally or alternatively be employed. It is also known to use similar techniques to prepare other oligonucleotides, such as the phosphorothioates and alkylated derivatives. 
     In the ligand-conjugated oligonucleotides and ligand-molecule bearing sequence-specific linked nucleosides of the present invention, the oligonucleotides and oligonucleosides may be assembled on a suitable DNA synthesizer utilizing standard nucleotide or nucleoside precursors, or nucleotide or nucleoside conjugate precursors that already bear the linking moiety, ligand-nucleotide or nucleoside-conjugate precursors that already bear the ligand molecule, or non-nucleoside ligand-bearing building blocks. 
     When using nucleotide-conjugate precursors that already bear a linking moiety, the synthesis of the sequence-specific linked nucleosides is typically completed, and the ligand molecule is then reacted with the linking moiety to form the ligand-conjugated oligonucleotide. In some embodiments, the oligonucleotides or linked nucleosides of the present invention are synthesized by an automated synthesizer using phosphoramidites derived from ligand-nucleoside conjugates in addition to the standard phosphoramidites and non-standard phosphoramidites that are commercially available and routinely used in oligonucleotide synthesis. 
     A. Lipid Conujugates 
     In one embodiment, 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, neproxin 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, and/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 a preferred embodiment, 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 another preferred embodiment, 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., glyciosylated 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, a 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 oligonucleotide further comprises a carbohydrate. The carbohydrate conjugated iRNA are advantageous for the in vivo delivery of nucleic acids, as well as compositions suitable for in vivo therapeutic use, as described herein. As used herein, “carbohydrate” refers to a compound which is either a carbohydrate per se made up of one or more monosaccharide units having at least 6 carbon atoms (which can be linear, branched or cyclic) with an oxygen, nitrogen, or sulfur atom bonded to each carbon atom; or a compound having as a part thereof a carbohydrate moiety made up of one or more monosaccharide units each having at least six carbon atoms (which can be linear, branched or cyclic), with an oxygen, nitrogen, or sulfur atom bonded to each carbon atom. Representative carbohydrates include the sugars (mono-, di-, tri- and oligosaccharides containing from about 4, 5, 6, 7, 8, or 9 monosaccharide units), and polysaccharides such as starches, glycogen, cellulose and polysaccharide gums. Specific monosaccharides include C5 and above (e.g., C5, C6, C7, or C8) sugars; di- and trisaccharides include sugars having two or three monosaccharide units (e.g., C5, C6, C7, or C8). 
     In one embodiment, 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 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. 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. The hairpin loop may also be formed by an extended overhang in one strand of the duplex. 
     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 and/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 NR8, C(O), C(O)NH, SO, SO 2 , SO 2 NH or a chain of atoms, such as, but not limited to, substituted or unsubstituted alkyl, substituted or unsubstituted alkenyl, substituted or unsubstituted alkynyl, arylalkyl, arylalkenyl, arylalkynyl, heteroarylalkyl, heteroarylalkenyl, heteroarylalkynyl, heterocyclylalkyl, heterocyclylalkenyl, heterocyclylalkynyl, aryl, heteroaryl, heterocyclyl, cycloalkyl, cycloalkenyl, alkylarylalkyl, alkylarylalkenyl, alkylarylalkynyl, alkenylarylalkyl, alkenylarylalkenyl, alkenylarylalkynyl, alkynylarylalkyl, alkynylarylalkenyl, alkynylarylalkynyl, alkylheteroarylalkyl, alkylheteroarylalkenyl, alkylheteroarylalkynyl, alkenylheteroarylalkyl, alkenylheteroarylalkenyl, alkenylheteroarylalkynyl, alkynylheteroarylalkyl, alkynylheteroarylalkenyl, alkynylheteroarylalkynyl, alkylheterocyclylalkyl, alkylheterocyclylalkenyl, alkylhererocyclylalkynyl, alkenylheterocyclylalkyl, alkenylheterocyclylalkenyl, alkenylheterocyclylalkynyl, alkynylheterocyclylalkyl, alkynylheterocyclylalkenyl, alkynylheterocyclylalkynyl, alkylaryl, alkenylaryl, alkynylaryl, alkylheteroaryl, alkenylheteroaryl, alkynylhereroaryl, which one or more methylenes can be interrupted or terminated by O, S, S(O), SO 2 , N(R8), C(O), substituted or unsubstituted aryl, substituted or unsubstituted heteroaryl, substituted or unsubstituted heterocyclic; where R8 is hydrogen, acyl, aliphatic or substituted aliphatic. In one embodiment, the linker is between about 1-24 atoms, 2-24, 3-24, 4-24, 5-24, 6-24, 6-18, 7-18, 8-18 atoms, 7-17, 8-17, 6-16, 7-17, 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 about 100 times faster in a target cell or under a first reference condition (which can, e.g., be selected to mimic or represent intracellular conditions) than in the blood of a subject, or under a second reference condition (which can, e.g., be selected to mimic or represent conditions found in the blood or serum). 
     Cleavable linking groups are susceptible to cleavage agents, e.g., pH, redox potential or the presence of degradative molecules. Generally, cleavage agents are more prevalent or found at higher levels or activities inside cells than in serum or blood. Examples of such degradative agents include: redox agents which are selected for particular substrates or which have no substrate specificity, including, e.g., oxidative or reductive enzymes or reductive agents such as mercaptans, present in cells, that can degrade a redox cleavable linking group by reduction; esterases; endosomes or agents that can create an acidic environment, e.g., those that result in a pH of five or lower; enzymes that can hydrolyze or degrade an acid cleavable linking group by acting as a general acid, peptidases (which can be substrate specific), and phosphatases. 
     A cleavable linkage group, such as a disulfide bond can be susceptible to pH. The pH of human serum is 7.4, while the average intracellular pH is slightly lower, ranging from about 7.1-7.3. Endosomes have a more acidic pH, in the range of 5.5-6.0, and lysosomes have an even more acidic pH at around 5.0. Some linkers will have a cleavable linking group that is cleaved at a preferred pH, thereby releasing a cationic lipid from the ligand inside the cell, or into the desired compartment of the cell. 
     A linker can include a cleavable linking group that is cleavable by a particular enzyme. The type of cleavable linking group incorporated into a linker can depend on the cell to be targeted. For example, a liver-targeting ligand can be linked to a cationic lipid through a linker that includes an ester group. Liver cells are rich in esterases, and therefore the linker will be cleaved more efficiently in liver cells than in cell types that are not esterase-rich. Other cell-types rich in esterases include cells of the lung, renal cortex, and testis. 
     Linkers that contain peptide bonds can be used when targeting cell types rich in peptidases, such as liver cells and synoviocytes. 
     In general, the suitability of a candidate cleavable linking group can be evaluated by testing the ability of a degradative agent (or condition) to cleave the candidate linking group. It will also be desirable to also test the candidate cleavable linking group for the ability to resist cleavage in the blood or when in contact with other non-target tissue. Thus, one can determine the relative susceptibility to cleavage between a first and a second condition, where the first is selected to be indicative of cleavage in a target cell and the second is selected to be indicative of cleavage in other tissues or biological fluids, e.g., blood or serum. The evaluations can be carried out in cell free systems, in cells, in cell culture, in organ or tissue culture, or in whole animals. It can be useful to make initial evaluations in cell-free or culture conditions and to confirm by further evaluations in whole animals. In preferred embodiments, useful candidate compounds are cleaved at least about 2, 4, 10, 20, 30, 40, 50, 60, 70, 80, 90, or about 100 times faster in the cell (or under in vitro conditions selected to mimic intracellular conditions) as compared to blood or serum (or under in vitro conditions selected to mimic extracellular conditions). 
     i. Redox Cleavable Linking Groups 
     In one embodiment, 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 another embodiment, 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—, —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 another embodiment, a cleavable linker comprises an acid cleavable linking group. An acid cleavable linking group is a linking group that is cleaved under acidic conditions. In preferred embodiments acid cleavable linking groups are cleaved in an acidic environment with a pH of about 6.5 or lower (e.g., about 6.0, 5.75, 5.5, 5.25, 5.0, or lower), or by agents such as enzymes that can act as a general acid. In a cell, specific low pH organelles, such as endosomes and lysosomes can provide a cleaving environment for acid cleavable linking groups. Examples of acid cleavable linking groups include but are not limited to hydrazones, esters, and esters of amino acids. Acid cleavable groups can have the general formula —C═NN—, C(O)O, or —OC(O). A preferred embodiment is when the carbon attached to the oxygen of the ester (the alkoxy group) is an aryl group, substituted alkyl group, or tertiary alkyl group such as dimethyl pentyl or t-butyl. These candidates can be evaluated using methods analogous to those described above. 
     iv. Ester-Based Linking Groups 
     In another embodiment, 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 another embodiment, a cleavable linker comprises a peptide-based cleavable linking group. A peptide-based cleavable linking group is cleaved by enzymes such as peptidases and proteases in cells. Peptide-based cleavable linking groups are peptide bonds formed between amino acids to yield oligopeptides (e.g., dipeptides, tripeptides, etc.) and polypeptides. Peptide-based cleavable groups do not include the amide group (—C(O)NH—). The amide group can be formed between any alkylene, alkenylene, or alkynelene. A peptide bond is a special type of amide bond formed between amino acids to yield peptides and proteins. The peptide based cleavage group is generally limited to the peptide bond (i.e., the amide bond) formed between amino acids yielding peptides and proteins and does not include the entire amide functional group. Peptide-based cleavable linking groups have the general formula —NHCHRAC(O)NHCHRBC(O)—, where RA and RB are the R groups of the two adjacent amino acids. These candidates can be evaluated using methods analogous to those described above. 
     In one embodiment, 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 (XXXII)-(XXXV): 
     
       
         
         
             
             
         
       
     
     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,
 
     
       
         
         
             
             
         
       
     
     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 (XXXVI): 
     
       
         
         
             
             
         
       
         
         
           
             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 and 5,688,941; 6,294,664; 6,320,017; 6,576,752; 6,783,931; 6,900,297; 7,037,646; 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 dsRNAs, which 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, and/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 an RNAs bearing an amino linker 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 having a glycogen storage disease (GSD), e.g., type Ia GSD) 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. The non-specific effects of an iRNA can be minimized by local administration, for example, by direct injection or implantation into a tissue or topically administering the preparation. Local administration to a treatment site maximizes local concentration of the agent, limits the exposure of the agent to systemic tissues that can otherwise be harmed by the agent or that can degrade the agent, and permits a lower total dose of the iRNA molecule to be administered. Several studies have shown successful knockdown of gene products when an iRNA is administered locally. For example, intraocular delivery of a VEGF dsRNA by intravitreal injection in cynomolgus monkeys (Tolentino, M J. et al., (2004)  Retina  24:132-138) and subretinal injections in mice (Reich, S J. et al. (2003)  Mol. Vis.  9:210-216) were both shown to prevent neovascularization in an experimental model of age-related macular degeneration. In addition, direct intratumoral injection of a dsRNA in mice reduces tumor volume (Pille, J. et al. (2005)  Mol. Ther.  11:267-274) and can prolong survival of tumor-bearing mice (Kim, W J. et al., (2006)  Mol. Ther.  14:343-350; Li, S. et al., (2007)  Mol. Ther.  15:515-523). RNA interference has also shown success with local delivery to the CNS by direct injection (Dorn, G. et al., (2004)  Nucleic Acids  32:e49; Tan, P H. et al. (2005)  Gene Ther.  12:59-66; Makimura, H. et a.l (2002)  BMC Neurosci.  3:18; Shishkina, G T., et al. (2004)  Neuroscience  129:521-528; Thakker, E R., et al. (2004)  Proc. Natl. Acad. Sci. U.S.A.  101:17270-17275; Akaneya, Y., et al. (2005)  J Neurophysiol.  93:594-602) and to the lungs by intranasal administration (Howard, K A. et al., (2006)  Mol. Ther.  14:476-484; Zhang, X. et al., (2004)  J. Biol. Chem.  279:10677-10684; Bitko, V. et al., (2005)  Nat. Med.  11:50-55). For administering an iRNA systemically for the treatment of a disease, the RNA can be modified or alternatively delivered using a drug delivery system; both methods act to prevent the rapid degradation of the dsRNA by endo- and exo-nucleases in vivo. Modification of the RNA or the pharmaceutical carrier can also permit targeting of the iRNA composition 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). Conjugation of an iRNA to an aptamer has been shown to inhibit tumor growth and mediate tumor regression in a mouse model of prostate cancer (McNamara, J O. et al., (2006)  Nat. Biotechnol.  24:1005-1015). In an alternative embodiment, the 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), Oligofectamine, “solid nucleic acid lipid particles” (Zimmermann, T S. et al., (2006)  Nature  441:111-114), cardiolipin (Chien, P Y. et al., (2005)  Cancer Gene Ther.  12:321-328; Pal, A. et al., (2005)  Intl 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 GCK 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., (1995)  Proc. Natl. Acad. Sci. USA  92:1292). 
     The individual strand or strands of an iRNA can be transcribed from a promoter on an expression vector. Where two separate strands are to be expressed to generate, for example, a dsRNA, two separate expression vectors can be co-introduced (e.g., by transfection or infection) into a target cell. Alternatively each individual strand of a dsRNA can be transcribed by promoters both of which are located on the same expression plasmid. In one embodiment, a dsRNA is expressed as inverted repeat polynucleotides joined by a linker polynucleotide sequence such that the dsRNA has a stem and loop structure. 
     iRNA expression vectors are generally DNA plasmids or viral vectors. Expression vectors compatible with eukaryotic cells, preferably those compatible with vertebrate cells, can be used to produce recombinant constructs for the expression of an iRNA as described herein. Eukaryotic cell expression vectors are well known in the art and are available from a number of commercial sources. Typically, such vectors are provided containing convenient restriction sites for insertion of the desired nucleic acid segment. Delivery of iRNA expressing vectors can be systemic, such as by intravenous or intramuscular administration, by administration to target cells ex-planted from the patient followed by reintroduction into the patient, or by any other means that allows for introduction into a desired target cell. 
     Viral vector systems which can be utilized with the methods and compositions described herein include, but are not limited to, (a) adenovirus vectors; (b) retrovirus vectors, including but not limited to lentiviral vectors, moloney murine leukemia virus, etc.; (c) adeno-associated virus vectors; (d) herpes simplex virus vectors; (e) SV 40 vectors; (0 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 treating a disease or disorder associated with the expression or activity of a GCK gene, such as, a glycogen storage disease (GSD), e.g., type Ia GSD, or one or more signs or symptoms of GSD, such as hypoglycemia, lactic acidosis, hyperuricemia, hyperlipidemia, hepatomegaly, kidney disease as a result of glycogen accumulation, hunger, jitteriness, lethargy, apnea, seizures, diaphoresis, confusion, headaches, dizziness, unusual mood or behavior changes, loss of consciousness, coma, muscle cramps, bleeding diathesis, short stauture, osteoporosis, delayed puberty, gout, renal disease, systemic hypertension, pulmonary hypertension, hepatic adenomas, pancreatitis, anemia, vitamin D deficiency, polycystic ovaries, irregular menstrual cycles, menorrhagia, and eruptive xanthomata. 
     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 intravenous (IV) or for subcutaneous delivery. Another example is compositions that are formulated for direct delivery into the liver, e.g., by infusion into the liver, such as by continuous pump infusion. 
     The pharmaceutical compositions of the invention may be administered in dosages sufficient to inhibit expression of a GCK 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 regimine may include administration of a therapeutic amount of iRNA on a regular basis, such as every other day or once a year. In certain embodiments, the iRNA is administered about once per month to about once per quarter (i.e., about once every three months). 
     After an initial treatment regimen, the treatments can be administered on a less frequent basis. 
     The skilled artisan will appreciate that certain factors can influence the dosage and timing required to effectively treat a subject, including but not limited to the severity of the disease or disorder, previous treatments, the general health and/or age of the subject, and other diseases present. Moreover, treatment of a subject with a therapeutically effective amount of a composition can include a single treatment or a series of treatments. Estimates of effective dosages and in vivo half-lives for the individual iRNAs encompassed by the invention can be made using conventional methodologies or on the basis of in vivo testing using an appropriate animal model, as described elsewhere herein. 
     Advances in mouse genetics have generated a number of mouse models for the study of various human diseases, such as disorders of excess glucose that would benefit from reduction in the expression of GCK. 
     The pharmaceutical compositions of the present invention can be administered in a number of ways depending upon whether local or systemic treatment is desired and upon the area to be treated. Administration can be topical (e.g., by a transdermal patch), pulmonary, e.g., by inhalation or insufflation of powders or aerosols, including by nebulizer; intratracheal, intranasal, epidermal and transdermal, oral or parenteral. Parenteral administration includes intravenous, intraarterial, subcutaneous, intraperitoneal or intramuscular injection or infusion; subdermal, e.g., via an implanted device; or intracranial, e.g., by intraparenchymal, intrathecal or intraventricular, administration. 
     The iRNA can be delivered in a manner to target a particular tissue, such as the liver (e.g., the hepatocytes of the liver). 
     Pharmaceutical compositions and formulations for topical administration can include transdermal patches, ointments, lotions, creams, gels, drops, suppositories, sprays, liquids, and powders. Conventional pharmaceutical carriers, aqueous, powder, or oily bases, thickeners and the like can be necessary or desirable. Coated condoms, gloves and the like can also be useful. Suitable topical formulations include those in which the iRNAs featured in the invention are in admixture with a topical delivery agent such as lipids, liposomes, fatty acids, fatty acid esters, steroids, chelating agents, and surfactants. Suitable lipids and liposomes include neutral (e.g., dioleoylphosphatidyl DOPE ethanolamine, dimyristoylphosphatidyl choline DMPC, distearolyphosphatidyl choline), negative (e.g., dimyristoylphosphatidyl glycerol DMPG), and cationic (e.g., dioleoyltetramethylaminopropyl DOTAP and dioleoylphosphatidyl ethanolamine DOTMA). iRNAs featured in the invention can be encapsulated within liposomes or can form complexes thereto, in particular to cationic liposomes. Alternatively, iRNAs can be complexed to lipids, in particular to cationic lipids. Suitable fatty acids and esters include but are not limited to arachidonic acid, oleic acid, eicosanoic acid, lauric acid, caprylic acid, capric acid, myristic acid, palmitic acid, stearic acid, linoleic acid, linolenic acid, dicaprate, tricaprate, monoolein, dilaurin, glyceryl 1-monocaprate, 1-dodecylazacycloheptan-2-one, an acylcarnitine, an acylcholine, or a C1-20 alkyl ester (e.g., isopropylmyristate IPM), monoglyceride or diglyceride; or pharmaceutically acceptable salt thereof. Topical formulations are described in detail in U.S. Pat. No. 6,747,014, which is incorporated herein by reference. 
     A. iRNA Formulations Comprising Membranous Molecular Assemblies 
     An iRNA for use in the compositions and methods of the invention can be formulated for delivery in a membranous molecular assembly, e.g., a liposome or a micelle. As used herein, the term “liposome” refers to a vesicle composed of amphiphilic lipids arranged in at least one bilayer, e.g., one bilayer or a plurality of bilayers. Liposomes include unilamellar and multilamellar vesicles that have a membrane formed from a lipophilic material and an aqueous interior. The aqueous portion contains the iRNA composition. The lipophilic material isolates the aqueous interior from an aqueous exterior, which typically does not include the iRNA composition, although in some examples, it may. Liposomes are useful for the transfer and delivery of active ingredients to the site of action. Because the liposomal membrane is structurally similar to biological membranes, when liposomes are applied to a tissue, the liposomal bilayer fuses with bilayer of the cellular membranes. As the merging of the liposome and cell progresses, the internal aqueous contents that include the iRNA are delivered into the cell where the iRNA can specifically bind to a target RNA and can mediate RNAi. In some cases the liposomes are also specifically targeted, e.g., to direct the iRNA to particular cell types. 
     A liposome containing a RNAi agent can be prepared by a variety of methods. In one example, the lipid component of a liposome is dissolved in a detergent so that micelles are formed with the lipid component. For example, the lipid component can be an amphipathic cationic lipid or lipid conjugate. The detergent can have a high critical micelle concentration and may be nonionic. Exemplary detergents include cholate, CHAPS, octylglucoside, deoxycholate, and lauroyl sarcosine. The RNAi agent preparation is then added to the micelles that include the lipid component. The cationic groups on the lipid interact with the RNAi agent and condense around the RNAi agent to form a liposome. After condensation, the detergent is removed, e.g., by dialysis, to yield a liposomal preparation of RNAi agent. 
     If necessary a carrier compound that assists in condensation can be added during the condensation reaction, e.g., by controlled addition. For example, the carrier compound can be a polymer other than a nucleic acid (e.g., spermine or spermidine). pH can also adjusted to favor condensation. 
     Methods for producing stable polynucleotide delivery vehicles, which incorporate a polynucleotide/cationic lipid complex as structural components of the delivery vehicle, are further described in, e.g., WO 96/37194, the entire contents of which are incorporated herein by reference. Liposome formation can also include one or more aspects of exemplary methods described in Felgner, P. L. et al., (1987)  Proc. Natl. Acad. Sci. USA  8:7413-7417; U.S. Pat. Nos. 4,897,355; 5,171,678; Bangham et al., (1965)  M Mol. Biol.  23:238; Olson et al., (1979)  Biochim. Biophys. Acta  557:9; Szoka et al., (1978)  Proc. Natl. Acad. Sci.  75: 4194; Mayhew et al., (1984)  Biochim. Biophys. Acta  775:169; Kim et al., (1983)  Biochim. Biophys. Acta  728:339; and Fukunaga et al., (1984)  Endocrinol.  115:757. Commonly used techniques for preparing lipid aggregates of appropriate size for use as delivery vehicles include sonication and freeze-thaw plus extrusion (see, e.g., Mayer et al., (1986)  Biochim. Biophys. Acta  858:161. Microfluidization can be used when consistently small (50 to 200 nm) and relatively uniform aggregates are desired (Mayhew et al., (1984)  Biochim. Biophys. Acta  775:169. These methods are readily adapted to packaging RNAi agent preparations into liposomes. 
     Liposomes fall into two broad classes. Cationic liposomes are positively charged liposomes which interact with the negatively charged nucleic acid molecules to form a stable complex. The positively charged nucleic acid/liposome complex binds to the negatively charged cell surface and is internalized in an endosome. Due to the acidic pH within the endosome, the liposomes are ruptured, releasing their contents into the cell cytoplasm (Wang et al. (1987)  Biochem. Biophys. Res. Commun.,  147:980-985). 
     Liposomes, which are pH-sensitive or negatively charged, entrap nucleic acids rather than complex with them. Since both the nucleic acid and the lipid are similarly charged, repulsion rather than complex formation occurs. Nevertheless, some nucleic acid is entrapped within the aqueous interior of these liposomes. pH sensitive liposomes have been used to deliver nucleic acids encoding the thymidine kinase gene to cell monolayers in culture. Expression of the exogenous gene was detected in the target cells (Zhou et al. (1992)  Journal of Controlled Release,  19:269-274). 
     One major type of liposomal composition includes phospholipids other than naturally-derived phosphatidylcholine. Neutral liposome compositions, for example, can be formed from dimyristoyl phosphatidylcholine (DMPC) or dipalmitoyl phosphatidylcholine (DPPC). Anionic liposome compositions generally are formed from dimyristoyl phosphatidylglycerol, while anionic fusogenic liposomes are formed primarily from dioleoyl phosphatidylethanolamine (DOPE). Another type of liposomal composition is formed from phosphatidylcholine (PC) such as, for example, soybean PC, and egg PC. Another type is formed from mixtures of phospholipid and/or phosphatidylcholine and/or cholesterol. 
     Examples of other methods to introduce liposomes into cells in vitro and in vivo include U.S. Pat. Nos. 5,283,185; 5,171,678; WO 94/00569; WO 93/24640; WO 91/16024; Felgner, (1994)  J Biol. Chem.  269:2550; Nabel, (1993)  Proc. Natl. Acad. Sci.  90:11307; Nabel, (1992)  Human Gene Ther.  3:649; Gershon, (1993)  Biochem.  32:7143; and Strauss, (1992)  EMBO J.  11:417. 
     Non-ionic liposomal systems have also been examined to determine their utility in the delivery of drugs to the skin, in particular systems comprising non-ionic surfactant and cholesterol. Non-ionic liposomal formulations comprising Novasome™ I (glyceryl dilaurate/cholesterol/polyoxyethylene-10-stearyl ether) and Novasome™ II (glyceryl distearate/cholesterol/polyoxyethylene-10-stearyl ether) were used to deliver cyclosporin-A into the dermis of mouse skin. Results indicated that such non-ionic liposomal systems were effective in facilitating the deposition of cyclosporine A into different layers of the skin (Hu et al., (1994)  S.T.P.Pharma. Sci.,  4(6):466). 
     Liposomes also include “sterically stabilized” liposomes, a term which, as used herein, refers to liposomes comprising one or more specialized lipids that, when incorporated into liposomes, result in enhanced circulation lifetimes relative to liposomes lacking such specialized lipids. Examples of sterically stabilized liposomes are those in which part of the vesicle-forming lipid portion of the liposome (A) comprises one or more glycolipids, such as monosialoganglioside G M1 , or (B) is derivatized with one or more hydrophilic polymers, such as a polyethylene glycol (PEG) moiety. While not wishing to be bound by any particular theory, it is thought in the art that, at least for sterically stabilized liposomes containing gangliosides, sphingomyelin, or PEG-derivatized lipids, the enhanced circulation half-life of these sterically stabilized liposomes derives from a reduced uptake into cells of the reticuloendothelial system (RES) (Allen et al., (1987)  FEBS Letters,  223:42; Wu et al., (1993)  Cancer Research,  53:3765). 
     Various liposomes comprising one or more glycolipids are known in the art. Papahadjopoulos et al. ( Ann. N.Y. Acad. Sci ., (1987), 507:64) reported the ability of monosialoganglioside G M1 , galactocerebroside sulfate and phosphatidylinositol to improve blood half-lives of liposomes. These findings were expounded upon by Gabizon et al. ( Proc. Natl. Acad. Sci. USA ., (1988), 85, 6949). U.S. Pat. No. 4,837,028 and WO 88/04924, both to Allen et al., disclose liposomes comprising (1) sphingomyelin and (2) the ganglioside G M1  or a galactocerebroside sulfate ester. U.S. Pat. No. 5,543,152 (Webb et al.) discloses liposomes comprising sphingomyelin. Liposomes comprising 1,2-sn-dimyristoylphosphatidylcholine are disclosed in WO 97/13499 (Lim et al). 
     In one embodiment, cationic liposomes are used. Cationic liposomes possess the advantage of being able to fuse to the cell membrane. Non-cationic liposomes, although not able to fuse as efficiently with the plasma membrane, are taken up by macrophages in vivo and can be used to deliver RNAi agents to macrophages. 
     Further advantages of liposomes include: liposomes obtained from natural phospholipids are biocompatible and biodegradable; liposomes can incorporate a wide range of water and lipid soluble drugs; liposomes can protect encapsulated RNAi agents in their internal compartments from metabolism and degradation (Rosoff, in “Pharmaceutical Dosage Forms,” Lieberman, Rieger and Banker (Eds.), 1988, volume 1, p. 245). Important considerations in the preparation of liposome formulations are the lipid surface charge, vesicle size and the aqueous volume of the liposomes. 
     A positively charged synthetic cationic lipid, N-[1-(2,3-dioleyloxy)propyl]-N,N,N-trimethylammonium chloride (DOTMA) can be used to form small liposomes that interact spontaneously with nucleic acid to form lipid-nucleic acid complexes which are capable of fusing with the negatively charged lipids of the cell membranes of tissue culture cells, resulting in delivery of RNAi agent (see, e.g., Felgner, P. L. et al., (1987)  Proc. Natl. Acad. Sci. USA  8:7413-7417, and U.S. Pat. No. 4,897,355 for a description of DOTMA and its use with DNA). 
     A DOTMA analogue, 1,2-bis(oleoyloxy)-3-(trimethylammonia)propane (DOTAP) can be used in combination with a phospholipid to form DNA-complexing vesicles. Lipofectin™ (Bethesda Research Laboratories, Gaithersburg, Md.) is an effective agent for the delivery of highly anionic nucleic acids into living tissue culture cells that comprise positively charged DOTMA liposomes which interact spontaneously with negatively charged polynucleotides to form complexes. When enough positively charged liposomes are used, the net charge on the resulting complexes is also positive. Positively charged complexes prepared in this way spontaneously attach to negatively charged cell surfaces, fuse with the plasma membrane, and efficiently deliver functional nucleic acids into, for example, tissue culture cells. Another commercially available cationic lipid, 1,2-bis(oleoyloxy)-3,3-(trimethylammonia)propane (“DOTAP”) (Boehringer Mannheim™, Indianapolis, Ind.) differs from DOTMA in that the oleoyl moieties are linked by ester, rather than ether linkages. 
     Other reported cationic lipid compounds include those that have been conjugated to a variety of moieties including, for example, carboxyspermine which has been conjugated to one of two types of lipids and includes compounds such as 5-carboxyspermylglycine dioctaoleoylamide (“DOGS”) (Transfectam™, Promega™, Madison, Wis.) and dipalmitoylphosphatidylethanolamine 5-carboxyspermyl-amide (“DPPES”) (see, e.g., U.S. Pat. No. 5,171,678). 
     Another cationic lipid conjugate includes derivatization of the lipid with cholesterol (“DC-Chol”) which has been formulated into liposomes in combination with DOPE (See, Gao, X. and Huang, L., (1991)  Biochim. Biophys. Res. Commun.  179:280). Lipopolylysine, made by conjugating polylysine to DOPE, has been reported to be effective for transfection in the presence of serum (Zhou, X. et al., (1991)  Biochim. Biophys. Acta  1065:8). For certain cell lines, these liposomes containing conjugated cationic lipids, are said to exhibit lower toxicity and provide more efficient transfection than the DOTMA-containing compositions. Other commercially available cationic lipid products include DMRIE and DMRIE-HP (Vical™, La Jolla, Calif.) and Lipofectamine™ (DOSPA) (Life Technology™, Inc., Gaithersburg, Md.). Other cationic lipids suitable for the delivery of oligonucleotides are described in WO 98/39359 and WO 96/37194. 
     Liposomal formulations are particularly suited for topical administration, liposomes present several advantages over other formulations. Such advantages include reduced side effects related to high systemic absorption of the administered drug, increased accumulation of the administered drug at the desired target, and the ability to administer RNAi agent into the skin. In some implementations, liposomes are used for delivering RNAi agent to epidermal cells and also to enhance the penetration of RNAi agent into dermal tissues, e.g., into skin. For example, the liposomes can be applied topically. Topical delivery of drugs formulated as liposomes to the skin has been documented (see, e.g., Weiner et al., (1992)  Journal of Drug Targeting , vol. 2, 405-410 and du Plessis et al., (1992)  Antiviral Research,  18:259-265; Mannino, R. J. and Fould-Fogerite, S., (1998)  Biotechniques  6:682-690; Itani, T. et al., (1987)  Gene  56:267-276; Nicolau, C. et al. (1987)  Meth. Enzymol.  149:157-176; Straubinger, R. M. and Papahadjopoulos, D. (1983)  Meth. Enzymol.  101:512-527; Wang, C. Y. and Huang, L., (1987)  Proc. Natl. Acad. Sci. USA  84:7851-7855). 
     Non-ionic liposomal systems have also been examined to determine their utility in the delivery of drugs to the skin, in particular systems comprising non-ionic surfactant and cholesterol. Non-ionic liposomal formulations comprising Novasome I (glyceryl dilaurate/cholesterol/polyoxyethylene-10-stearyl ether) and Novasome II (glyceryl distearate/cholesterol/polyoxyethylene-10-stearyl ether) were used to deliver a drug into the dermis of mouse skin. Such formulations with RNAi agent are useful for treating a dermatological disorder. 
     Liposomes that include iRNA can be made highly deformable. Such deformability can enable the liposomes to penetrate through pore that are smaller than the average radius of the liposome. For example, transfersomes are a type of deformable liposomes. Transferosomes can be made by adding surface edge activators, usually surfactants, to a standard liposomal composition. Transfersomes that include RNAi agent can be delivered, for example, subcutaneously by infection in order to deliver RNAi agent to keratinocytes in the skin. In order to cross intact mammalian skin, lipid vesicles must pass through a series of fine pores, each with a diameter less than 50 nm, under the influence of a suitable transdermal gradient. In addition, due to the lipid properties, these transferosomes can be self-optimizing (adaptive to the shape of pores, e.g., in the skin), self-repairing, and can frequently reach their targets without fragmenting, and often self-loading. Transfersomes have been used to deliver serum albumin to the skin. The transfersome-mediated delivery of serum albumin has been shown to be as effective as subcutaneous injection of a solution containing serum albumin. 
     Other formulations amenable to the present invention are described in, for example, PCT Publication No. WO 2008/042973. 
     Transfersomes are yet another type of liposomes, and are highly deformable lipid aggregates which are attractive candidates for drug delivery vehicles. Transfersomes can be described as lipid droplets which are so highly deformable that they are easily able to penetrate through pores which are smaller than the droplet. Transfersomes are adaptable to the environment in which they are used, e.g., they are self-optimizing (adaptive to the shape of pores in the skin), self-repairing, frequently reach their targets without fragmenting, and often self-loading. To make transfersomes it is possible to add surface edge-activators, usually surfactants, to a standard liposomal composition. Transfersomes have been used to deliver serum albumin to the skin. The transfersome-mediated delivery of serum albumin has been shown to be as effective as subcutaneous injection of a solution containing serum albumin. 
     Surfactants find wide application in formulations such as emulsions (including microemulsions) and liposomes. The most common way of classifying and ranking the properties of the many different types of surfactants, both natural and synthetic, is by the use of the hydrophile/lipophile balance (HLB). The nature of the hydrophilic group (also known as the “head”) provides the most useful means for categorizing the different surfactants used in formulations (Rieger, in Pharmaceutical Dosage Forms, Marcel Dekker, Inc., New York, N.Y., 1988, p. 285). 
     If the surfactant molecule is not ionized, it is classified as a nonionic surfactant. Nonionic surfactants find wide application in pharmaceutical and cosmetic products and are usable over a wide range of pH values. In general their HLB values range from 2 to about 18 depending on their structure. Nonionic surfactants include nonionic esters such as ethylene glycol esters, propylene glycol esters, glyceryl esters, polyglyceryl esters, sorbitan esters, sucrose esters, and ethoxylated esters. Nonionic alkanolamides and ethers such as fatty alcohol ethoxylates, propoxylated alcohols, and ethoxylated/propoxylated block polymers are also included in this class. The polyoxyethylene surfactants are the most popular members of the nonionic surfactant class. 
     If the surfactant molecule carries a negative charge when it is dissolved or dispersed in water, the surfactant is classified as anionic. Anionic surfactants include carboxylates such as soaps, acyl lactylates, acyl amides of amino acids, esters of sulfuric acid such as alkyl sulfates and ethoxylated alkyl sulfates, sulfonates such as alkyl benzene sulfonates, acyl isethionates, acyl taurates and sulfosuccinates, and phosphates. The most important members of the anionic surfactant class are the alkyl sulfates and the soaps. 
     If the surfactant molecule carries a positive charge when it is dissolved or dispersed in water, the surfactant is classified as cationic. Cationic surfactants include quaternary ammonium salts and ethoxylated amines. The quaternary ammonium salts are the most used members of this class. 
     If the surfactant molecule has the ability to carry either a positive or negative charge, the surfactant is classified as amphoteric. Amphoteric surfactants include acrylic acid derivatives, substituted alkylamides, N-alkylbetaines, and phosphatides. 
     The use of surfactants in drug products, formulations and in emulsions has been reviewed (Rieger, in Pharmaceutical Dosage Forms, Marcel Dekker, Inc., New York, N.Y., 1988, p. 285). 
     The iRNA for use in the methods of the invention can also be provided as micellar formulations. “Micelles” are defined herein as a particular type of molecular assembly in which amphipathic molecules are arranged in a spherical structure such that all the hydrophobic portions of the molecules are directed inward, leaving the hydrophilic portions in contact with the surrounding aqueous phase. The converse arrangement exists if the environment is hydrophobic. 
     A mixed micellar formulation suitable for delivery through transdermal membranes may be prepared by mixing an aqueous solution of the siRNA composition, an alkali metal C 8  to C 22  alkyl sulphate, and a micelle forming compounds. Exemplary micelle forming compounds include lecithin, hyaluronic acid, pharmaceutically acceptable salts of hyaluronic acid, glycolic acid, lactic acid, chamomile extract, cucumber extract, oleic acid, linoleic acid, linolenic acid, monoolein, monooleates, monolaurates, borage oil, evening of primrose oil, menthol, trihydroxy oxo cholanyl glycine and pharmaceutically acceptable salts thereof, glycerin, polyglycerin, lysine, polylysine, triolein, polyoxyethylene ethers and analogues thereof, polidocanol alkyl ethers and analogues thereof, chenodeoxycholate, deoxycholate, and mixtures thereof. The micelle forming compounds may be added at the same time or after addition of the alkali metal alkyl sulphate. Mixed micelles will form with substantially any kind of mixing of the ingredients but vigorous mixing in order to provide smaller size micelles. 
     In one method a first micellar composition is prepared which contains the siRNA composition and at least the alkali metal alkyl sulphate. The first micellar composition is then mixed with at least three micelle forming compounds to form a mixed micellar composition. In another method, the micellar composition is prepared by mixing the siRNA composition, the alkali metal alkyl sulphate and at least one of the micelle forming compounds, followed by addition of the remaining micelle forming compounds, with vigorous mixing. 
     Phenol and/or m-cresol may be added to the mixed micellar composition to stabilize the formulation and protect against bacterial growth. Alternatively, phenol and/or m-cresol may be added with the micelle forming ingredients. An isotonic agent such as glycerin may also be added after formation of the mixed micellar composition. 
     For delivery of the micellar formulation as a spray, the formulation can be put into an aerosol dispenser and the dispenser is charged with a propellant. The propellant, which is under pressure, is in liquid form in the dispenser. The ratios of the ingredients are adjusted so that the aqueous and propellant phases become one, i.e., there is one phase. If there are two phases, it is necessary to shake the dispenser prior to dispensing a portion of the contents, e.g., through a metered valve. The dispensed dose of pharmaceutical agent is propelled from the metered valve in a fine spray. 
     Propellants may include hydrogen-containing chlorofluorocarbons, hydrogen-containing fluorocarbons, dimethyl ether, and diethyl ether. In certain embodiments, HFA 134a (1,1,1,2 tetrafluoroethane) may be used. 
     The specific concentrations of the essential ingredients can be determined by relatively straightforward experimentation. For absorption through the oral cavities, it is often desirable to increase, e.g., at least double or triple, the dosage for through injection or administration through the gastrointestinal tract. 
     B. Lipid Particles 
     iRNAs, e.g., dsRNAs of in the invention may be fully encapsulated in a lipid formulation, e.g., a LNP, or other nucleic acid-lipid particle. 
     As used herein, the term “LNP” refers to a stable nucleic acid-lipid particle. LNPs typically contain a cationic lipid, a non-cationic lipid, and a lipid that prevents aggregation of the particle (e.g., a PEG-lipid conjugate). LNPs are extremely useful for systemic applications, as they exhibit extended circulation lifetimes following intravenous (i.v.) injection and accumulate at distal sites (e.g., sites physically separated from the administration site). LNPs include “pSPLP,” which include an encapsulated condensing agent-nucleic acid complex as set forth in PCT Publication No. WO 00/03683. The particles of the present invention typically have a mean diameter of about 50 nm to about 150 nm, more typically about 60 nm to about 130 nm, more typically about 70 nm to about 110 nm, most typically about 70 nm to about 90 nm, and are substantially nontoxic. In addition, the nucleic acids when present in the nucleic acid-lipid particles of the present invention are resistant in aqueous solution to degradation with a nuclease. Nucleic acid-lipid particles and their method of preparation are disclosed in, e.g., U.S. Pat. Nos. 5,976,567; 5,981,501; 6,534,484; 6,586,410; 6,815,432; U.S. Publication No. 2010/0324120 and PCT Publication No. WO 96/40964. 
     In one embodiment, the lipid to drug ratio (mass/mass ratio) (e.g., lipid to dsRNA ratio) will be in the range of from about 1:1 to about 50:1, from about 1:1 to about 25:1, from about 3:1 to about 15:1, from about 4:1 to about 10:1, from about 5:1 to about 9:1, or about 6:1 to about 9:1. Ranges intermediate to the above recited ranges are also contemplated to be part of the invention. 
     The cationic lipid can be, for example, N,N-dioleyl-N,N-dimethylammonium chloride (DODAC), N,N-distearyl-N,N-dimethylammonium bromide (DDAB), N—(I-(2,3-dioleoyloxy)propyl)-N,N,N-trimethylammonium chloride (DOTAP), N—(I-(2,3-dioleyloxy)propyl)-N,N,N-trimethylammonium chloride (DOTMA), N,N-dimethyl-2,3-dioleyloxy)propylamine (DODMA), 1,2-DiLinoleyloxy-N,N-dimethylaminopropane (DLinDMA), 1,2-Dilinolenyloxy-N,N-dimethylaminopropane (DLenDMA), 1,2-Dilinoleylcarbamoyloxy-3-dimethylaminopropane (DLin-C-DAP), 1,2-Dilinoleyoxy-3-(dimethylamino)acetoxypropane (DLin-DAC), 1,2-Dilinoleyoxy-3-morpholinopropane (DLin-MA), 1,2-Dilinoleoyl-3-dimethylaminopropane (DLinDAP), 1,2-Dilinoleylthio-3-dimethylaminopropane (DLin-S-DMA), 1-Linoleoyl-2-linoleyloxy-3-dimethylaminopropane (DLin-2-DMAP), 1,2-Dilinoleyloxy-3-trimethylaminopropane chloride salt (DLin-TMA.Cl), 1,2-Dilinoleoyl-3-trimethylaminopropane chloride salt (DLin-TAP.Cl), 1,2-Dilinoleyloxy-3-(N-methylpiperazino)propane (DLin-MPZ), or 3-(N,N-Dilinoleylamino)-1,2-propanediol (DLinAP), 3-(N,N-Dioleylamino)-1,2-propanedio (DOAP), 1,2-Dilinoleyloxo-3-(2-N,N-dimethylamino)ethoxypropane (DLin-EG-DMA), 1,2-Dilinolenyloxy-N,N-dimethylaminopropane (DLinDMA), 2,2-Dilinoleyl-4-dimethylaminomethyl-[1,3]-dioxolane (DLin-K-DMA) or analogs thereof, (3aR,5s,6aS)—N,N-dimethyl-2,2-di((9Z,12Z)-octadeca-9,12-dienyl)tetrahydro-3aH-cyclopenta[d][1,3]dioxol-5-amine (ALN100), (6Z,9Z,28Z,31Z)-heptatriaconta-6,9,28,31-tetraen-19-yl 4-(dimethylamino)butanoate (MC3), 1,1′-(2-(4-(2-((2-(bis(2-hydroxydodecyl)amino)ethyl)(2-hydroxydodecyl)amino)ethyl)piperazin-1-yl)ethylazanediyOdidodecan-2-ol (Tech G1), or a mixture thereof. The cationic lipid can comprise from about 20 mol % to about 50 mol % or about 40 mol % of the total lipid present in the particle. 
     In another embodiment, the compound 2,2-Dilinoleyl-4-dimethylaminoethyl-[1,3]-dioxolane can be used to prepare lipid-siRNA nanoparticles. Synthesis of 2,2-Dilinoleyl-4-dimethylaminoethyl-[1,3]-dioxolane is described in U.S. provisional patent application No. 61/107,998 filed on Oct. 23, 2008, which is herein incorporated by reference. 
     In one embodiment, the lipid-siRNA particle includes 40% 2, 2-Dilinoleyl-4-dimethylaminoethyl-[1,3]-dioxolane: 10% DSPC: 40% Cholesterol: 10% PEG-C-DOMG (mole percent) with a particle size of 63.0±20 nm and a 0.027 siRNA/Lipid Ratio. 
     The ionizable/non-cationic lipid can be an anionic lipid or a neutral lipid including, but not limited to, distearoylphosphatidylcholine (DSPC), dioleoylphosphatidylcholine (DOPC), dipalmitoylphosphatidylcholine (DPPC), dioleoylphosphatidylglycerol (DOPG), dipalmitoylphosphatidylglycerol (DPPG), dioleoyl-phosphatidylethanolamine (DOPE), palmitoyloleoylphosphatidylcholine (POPC), palmitoyloleoylphosphatidylethanolamine (POPE), dioleoyl-phosphatidylethanolamine 4-(N-maleimidomethyl)-cyclohexane-1-carboxylate (DOPE-mal), dipalmitoyl phosphatidyl ethanolamine (DPPE), dimyristoylphosphoethanolamine (DMPE), distearoyl-phosphatidyl-ethanolamine (DSPE), 16-O-monomethyl PE, 16-O-dimethyl PE, 18-1-trans PE, 1-stearoyl-2-oleoyl-phosphatidyethanolamine (SOPE), cholesterol; or a mixture thereof. The non-cationic lipid can be from about 5 mol % to about 90 mol %, about 10 mol %, or about 58 mol % if cholesterol is included, of the total lipid present in the particle. 
     The conjugated lipid that inhibits aggregation of particles can be, for example, a polyethyleneglycol (PEG)-lipid including, without limitation, a PEG-diacylglycerol (DAG), a PEG-dialkyloxypropyl (DAA), a PEG-phospholipid, a PEG-ceramide (Cer), or a mixture thereof. The PEG-DAA conjugate can be, for example, a PEG-dilauryloxypropyl (Ci 2 ), a PEG-dimyristyloxypropyl (Ci 4 ), a PEG-dipalmityloxypropyl (Ci 6 ), or a PEG-distearyloxypropyl (C] 8 ). The conjugated lipid that prevents aggregation of particles can be from 0 mol % to about 20 mol % or about 2 mol % of the total lipid present in the particle. 
     In some embodiments, the nucleic acid-lipid particle further includes cholesterol at, e.g., about 10 mol % to about 60 mol % or about 48 mol % of the total lipid present in the particle. 
     In one embodiment, the lipidoid ND98-4HC1 (MW 1487) (see US20090023673, filed Mar. 26, 2008, which is incorporated herein by reference), Cholesterol (Sigma-Aldrich™), and PEG-Ceramide C16 (Avanti™ Polar Lipids) can be used to prepare lipid-dsRNA nanoparticles (i.e., LNP01 particles). Stock solutions of each in ethanol can be prepared as follows: ND98, 133 mg/ml; cholesterol, 25 mg/ml, PEG-Ceramide C16, 100 mg/ml. The ND98, cholesterol, and PEG-Ceramide C16 stock solutions can then be combined in a, e.g., 42:48:10 molar ratio. The combined lipid solution can be mixed with aqueous dsRNA (e.g., in sodium acetate pH 5) such that the final ethanol concentration is about 35-45% and the final sodium acetate concentration is about 100-300 mM. Lipid-dsRNA nanoparticles typically form spontaneously upon mixing. Depending on the desired particle size distribution, the resultant nanoparticle mixture can be extruded through a polycarbonate membrane (e.g., 100 nm cut-off) using, for example, a thermobarrel extruder, such as Lipex Extruder (Northern Lipids, Inc). In some cases, the extrusion step can be omitted. Ethanol removal and simultaneous buffer exchange can be accomplished by, for example, dialysis or tangential flow filtration. Buffer can be exchanged with, for example, phosphate buffered saline (PBS) at about pH 7, e.g., about pH 6.9, about pH 7.0, about pH 7.1, about pH 7.2, about pH 7.3, or about pH 7.4. 
     
       
         
         
             
             
         
       
     
     LNP01 formulations are described, e.g., in International Application Publication No. WO 2008/042973, which is hereby incorporated by reference. 
     Additional exemplary lipid-dsRNA formulations are described in the table below. 
     
       
         
           
               
               
               
             
               
                   
                   
               
               
                   
                   
                 cationic lipid/non-cationic 
               
               
                   
                   
                 lipid/cholesterol/PEG-lipid 
               
               
                   
                   
                 conjugate 
               
               
                   
                 Ionizable/Cationic Lipid 
                 Lipid:siRNA ratio 
               
               
                   
                   
               
             
            
               
                   
               
            
           
           
               
               
               
            
               
                 SNALP-1 
                 l,2-Dilinolenyloxy-N,N- 
                 DLinDMA/DPPC/Cholesterol/PEG-cDMA 
               
               
                   
                 dimethylaminopropane (DLinDMA) 
                 (57.1/7.1/34.4/1.4) 
               
               
                   
                   
                 lipid:siRNA ~7:1 
               
               
                 2-XTC 
                 2,2-Dilinoleyl-4-dimethylaminoethyl-[1,3]- 
                 XTC/DPPC/Cholesterol/PEG-cDMA 
               
               
                   
                 dioxolane (XTC) 
                 57.1/7.1/34.4/1.4 
               
               
                   
                   
                 lipid:siRNA ~7:1 
               
               
                 LNP05 
                 2,2-Dilinoleyl-4-dimethylaminoethyl-[1,3]- 
                 XTC/DSPC/Cholesterol/PEG-DMG 
               
               
                   
                 dioxolane (XTC) 
                 57.5/7.5/31.5/3.5 
               
               
                   
                   
                 lipid:siRNA ~6:1 
               
               
                 LNP06 
                 2,2-Dilinoleyl-4-dimethylaminoethyl-[1,3]- 
                 XTC/DSPC/Cholesterol/PEG-DMG 
               
               
                   
                 dioxolane (XTC) 
                 57.5/7.5/31.5/3.5 
               
               
                   
                   
                 lipid:siRNA ~11:1 
               
               
                 LNP07 
                 2,2-Dilinoleyl-4-dimethylaminoethyl-[1,3]- 
                 XTC/DSPC/Cholesterol/PEG-DMG 
               
               
                   
                 dioxolane (XTC) 
                 60/7.5/31/1.5, 
               
               
                   
                   
                 lipid:siRNA ~6:1 
               
               
                 LNP08 
                 2,2-Dilinoleyl-4-dimethylaminoethyl-[1,3]- 
                 XTC/DSC/Cholesterol/PEG-DMG 
               
               
                   
                 dioxolane (XTC) 
                 60/7.5/31/1.5, 
               
               
                   
                   
                 lipid:siRNA ~11:1 
               
               
                 LNP09 
                 2,2-Dilinoleyl-4-dimethylaminoethyl-[1,3]- 
                 XTC/DSPC/Cholesterol/PEG-DMG 
               
               
                   
                 dioxolane (XTC) 
                 50/10/38.5/1.5 
               
               
                   
                   
                 Lipid:siRNA 10:1 
               
               
                 LNP10 
                 (3aR,5s,6aS)-N,N-dimethyl-2,2-di((9Z,12Z)- 
                 ALN100/DSPC/Cholesterol/PEG-DMG 
               
               
                   
                 octadeca-9,12-dienyl)tetrahydro-3aH- 
                 50/10/38.5/1.5 
               
               
                   
                 cyclopenta[d][1,3]dioxol-5-amine (ALN100) 
                 Lipid:siRNA 10:1 
               
               
                 LNP11 
                 (6Z,9Z,28Z,31Z)-heptatriaconta-6,9,28,31- 
                 MC-3/DSPC/Cholesterol/PEG-DMG 
               
               
                   
                 tetraen-19-yl 4-(dimethylamino)butanoate 
                 50/10/38.5/1.5 
               
               
                   
                 (MC3) 
                 Lipid:siRNA 10:1 
               
               
                 LNP12 
                 1,1′-(2-(4-(2-((2-(bis(2- 
                 Tech G1/DSPC/Cholesterol/PEG-DMG 
               
               
                   
                 hydroxydodecyl)amino)ethyl)(2- 
                 50/10/38.5/1.5 
               
               
                   
                 hydroxydodecyl)amino)ethyl)piperazin-1- 
                 Lipid:siRNA 10:1 
               
               
                   
                 yl)ethylazanediyl)didodecan-2-ol (Tech G1) 
               
               
                 LNP13 
                 XTC 
                 XTC/DSPC/Chol/PEG-DMG 
               
               
                   
                   
                 50/10/38.5/1.5 
               
               
                   
                   
                 Lipid:siRNA: 33:1 
               
               
                 LNP14 
                 MC3 
                 MC3/DSPC/Chol/PEG-DMG 
               
               
                   
                   
                 40/15/40/5 
               
               
                   
                   
                 Lipid:siRNA: 11:1 
               
               
                 LNP15 
                 MC3 
                 MC3/DSPC/Chol/PEG- 
               
               
                   
                   
                 DSG/GalNAc-PEG-DSG 
               
               
                   
                   
                 50/10/35/4.5/0.5 
               
               
                   
                   
                 Lipid:siRNA: 11:1 
               
               
                 LNP16 
                 MC3 
                 MC3/DSPC/Chol/PEG-DMG 
               
               
                   
                   
                 50/10/38.5/1.5 
               
               
                   
                   
                 Lipid:siRNA: 7:1 
               
               
                 LNP17 
                 MC3 
                 MC3/DSPC/Chol/PEG-DSG 
               
               
                   
                   
                 50/10/38.5/1.5 
               
               
                   
                   
                 Lipid:siRNA: 10:1 
               
               
                 LNP18 
                 MC3 
                 MC3/DSPC/Chol/PEG-DMG 
               
               
                   
                   
                 50/10/38.5/1.5 
               
               
                   
                   
                 Lipid:siRNA: 12:1 
               
               
                 LNP19 
                 MC3 
                 MC3/DSPC/Chol/PEG-DMG 
               
               
                   
                   
                 50/10/35/5 
               
               
                   
                   
                 Lipid:siRNA: 8:1 
               
               
                 LNP20 
                 MC3 
                 MC3/DSPC/Chol/PEG-DPG 
               
               
                   
                   
                 50/10/38.5/1.5 
               
               
                   
                   
                 Lipid:siRNA: 10:1 
               
               
                 LNP21 
                 C12-200 
                 C12-200/DSPC/Chol/PEG-DSG 
               
               
                   
                   
                 50/10/38.5/1.5 
               
               
                   
                   
                 Lipid:siRNA: 7:1 
               
               
                 LNP22 
                 XTC 
                 XTC/DSPC/Chol/PEG-DSG 
               
               
                   
                   
                 50/10/38.5/1.5 
               
               
                   
                   
                 Lipid:siRNA: 10:1 
               
               
                   
               
               
                 DSPC: distearoylphosphatidylcholine 
               
               
                 DPPC: dipalmitoylphosphatidylcholine 
               
               
                 PEG-DMG: PEG-didimyristoyl glycerol (C14-PEG, or PEG-C14) (PEG with avg mol wt of 2000) 
               
               
                 PEG-DSG: PEG-distyryl glycerol (C18-PEG, or PEG-C18) (PEG with avg mol wt of 2000) 
               
               
                 PEG-cDMA: PEG-carbamoyl-1,2-dimyristyloxypropylamine (PEG with avg mol wt of 2000) 
               
               
                 SNALP (l,2-Dilinolenyloxy-N,N-dimethylaminopropane (DLinDMA)) comprising formulations are described in International Publication No. WO2009/127060, filed Apr. 15. 2009. which is hereby incorporated by reference. 
               
               
                 XTC comprising formulations are described in, for example, PCT Publication No. WO 2010/088537, the entire contents of which are hereby incorporated herein by reference. 
               
               
                 MC3 comprising formulations are described, e.g., in U.S. Publication No. 2010/0324120, filed Jun. 10. 2010, the entire contents of which are hereby incorporated by reference. 
               
               
                 ALNY-100 comprising formulations are described in, for example, PCT Publication No. WO 2010/054406, the entire contents of which are hereby incorporated herein by reference. 
               
               
                 C12-200 comprising formulations are described in, for example, PCT Publication No. WO 2010/129709, the entire contents of which are hereby incorporated herein by reference. 
               
            
           
         
       
     
     Compositions and formulations for oral administration include powders or granules, microparticulates, nanoparticulates, suspensions, or solutions in water or non-aqueous media, capsules, gel capsules, sachets, tablets, or minitablets. Thickeners, flavoring agents, diluents, emulsifiers, dispersing aids, or binders can be desirable. In some embodiments, oral formulations are those in which dsRNAs featured in the invention are administered in conjunction with one or more penetration enhancer surfactants and chelators. Suitable surfactants include fatty acids and/or esters or salts thereof, bile acids and/or salts thereof. Suitable bile acids/salts include chenodeoxycholic acid (CDCA) and ursodeoxychenodeoxycholic acid (UDCA), cholic acid, dehydrocholic acid, deoxycholic acid, glucholic acid, glycholic acid, glycodeoxycholic acid, taurocholic acid, taurodeoxycholic acid, sodium tauro-24,25-dihydro-fusidate and sodium glycodihydrofusidate. Suitable fatty acids include arachidonic acid, undecanoic acid, oleic acid, lauric acid, caprylic acid, capric acid, myristic acid, palmitic acid, stearic acid, linoleic acid, linolenic acid, dicaprate, tricaprate, monoolein, dilaurin, glyceryl 1-monocaprate, 1-dodecylazacycloheptan-2-one, an acylcarnitine, an acylcholine, or a monoglyceride or a diglyceride; or a pharmaceutically acceptable salt thereof (e.g., sodium). In some embodiments, combinations of penetration enhancers are used, for example, fatty acids/salts in combination with bile acids/salts. One exemplary combination is the sodium salt of lauric acid, capric acid and UDCA. Further penetration enhancers include polyoxyethylene-9-lauryl ether, polyoxyethylene-20-cetyl ether. DsRNAs featured in the invention can be delivered orally, in granular form including sprayed dried particles, or complexed to form micro or nanoparticles. DsRNA complexing agents include polyamino acids; polyimines; polyacrylates; polyalkylacrylates, polyoxethanes, polyalkylcyanoacrylates; cationized gelatins, albumins, starches, acrylates, polyethyleneglycols (PEG) and starches; polyalkylcyanoacrylates; DEAE-derivatized polyimines, pollulans, celluloses and starches. Suitable complexing agents include chitosan, N-trimethylchitosan, poly-L-lysine, polyhistidine, polyornithine, polyspermines, protamine, polyvinylpyridine, polythiodiethylaminomethylethylene P(TDAE), polyaminostyrene (e.g., p-amino), poly(methylcyanoacrylate), poly(ethylcyanoacrylate), poly(butylcyanoacrylate), poly(isobutylcyanoacrylate), poly(isohexylcynaoacrylate), DEAE-methacrylate, DEAE-hexylacrylate, DEAE-acrylamide, DEAE-albumin and DEAE-dextran, polymethylacrylate, polyhexylacrylate, poly(D,L-lactic acid), poly(DL-lactic-co-glycolic acid (PLGA), alginate, and polyethyleneglycol (PEG). Oral formulations for dsRNAs and their preparation are described in detail in U.S. Pat. No. 6,887,906, US Publn. No. 20030027780, and U.S. Pat. No. 6,747,014, each of which is incorporated herein by reference. 
     Compositions and formulations for parenteral, intraparenchymal (into the brain), intrathecal, intraventricular, or intrahepatic administration can include sterile aqueous solutions which can also contain buffers, diluents and other suitable additives such as, but not limited to, penetration enhancers, carrier compounds, and other pharmaceutically acceptable carriers or excipients. 
     Pharmaceutical compositions of the present 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. Particularly preferred are formulations that target the liver when treating hepatic disorders such as hepatic carcinoma. 
     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 or finely divided solid carriers or both, and then, if necessary, shaping the product. 
     The compositions of the present invention can be formulated into any of many possible dosage forms such as, but not limited to, tablets, capsules, gel capsules, liquid syrups, soft gels, suppositories, and enemas. The compositions of the present invention can also be formulated as suspensions in aqueous, non-aqueous, or mixed media. Aqueous suspensions can further contain substances which increase the viscosity of the suspension including, for example, sodium carboxymethylcellulose, sorbitol and/or dextran. The suspension can also contain stabilizers. 
     C. 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 in either aqueous phase, oily phase or itself as a separate phase. Pharmaceutical excipients such as emulsifiers, stabilizers, dyes, and anti-oxidants can also be present in emulsions as needed. Pharmaceutical emulsions can also be multiple emulsions that are comprised of more than two phases such as, for example, in the case of oil-in-water-in-oil (o/w/o) and water-in-oil-in-water (w/o/w) emulsions. Such complex formulations often provide certain advantages that simple binary emulsions do not. Multiple emulsions in which individual oil droplets of an o/w emulsion enclose small water droplets constitute a w/o/w emulsion. Likewise a system of oil droplets enclosed in globules of water stabilized in an oily continuous phase provides an o/w/o emulsion. 
     Emulsions are characterized by little or no thermodynamic stability. Often, the dispersed or discontinuous phase of the emulsion is well dispersed into the external or continuous phase and maintained in this form through the means of emulsifiers or the viscosity of the formulation. Either of the phases of the emulsion can be a semisolid or a solid, as is the case of emulsion-style ointment bases and creams. Other means of stabilizing emulsions entail the use of emulsifiers that can be incorporated into either phase of the emulsion. Emulsifiers can broadly be classified into four categories: synthetic surfactants, naturally occurring emulsifiers, absorption bases, and finely dispersed solids (see e.g., Ansel&#39;s Pharmaceutical Dosage Forms and Drug Delivery Systems, Allen, L V., Popovich N G., and Ansel H C., 2004, Lippincott Williams &amp; Wilkins (8th ed.), New York, N.Y.; Idson, in Pharmaceutical Dosage Forms, Lieberman, Rieger and Banker (Eds.), 1988, Marcel Dekker, Inc., New York, N.Y., volume 1, p. 199). 
     Synthetic surfactants, also known as surface active agents, have found wide applicability in the formulation of emulsions and have been reviewed in the literature (see e.g., Ansel&#39;s Pharmaceutical Dosage Forms and Drug Delivery Systems, Allen, L V., Popovich N G., and Ansel H C., 2004, Lippincott Williams &amp; Wilkins (8th ed.), New York, N.Y.; Rieger, in Pharmaceutical Dosage Forms, Lieberman, Rieger and Banker (Eds.), 1988, Marcel Dekker, Inc., New York, N.Y., volume 1, p. 285; Idson, in Pharmaceutical Dosage Forms, Lieberman, Rieger and Banker (Eds.), Marcel Dekker, Inc., New York, N.Y., 1988, volume 1, p. 199). Surfactants are typically amphiphilic and comprise a hydrophilic and a hydrophobic portion. The ratio of the hydrophilic to the hydrophobic nature of the surfactant has been termed the hydrophile/lipophile balance (HLB) and is a valuable tool in categorizing and selecting surfactants in the preparation of formulations. Surfactants can be classified into different classes based on the nature of the hydrophilic group: nonionic, anionic, cationic and amphoteric (see e.g., Ansel&#39;s Pharmaceutical Dosage Forms and Drug Delivery Systems, Allen, L V., Popovich N G., and Ansel H C., 2004, Lippincott Williams &amp; Wilkins (8th ed.), New York, N.Y. Rieger, in Pharmaceutical Dosage Forms, Lieberman, Rieger and Banker (Eds.), 1988, Marcel Dekker, Inc., New York, N.Y., volume 1, p. 285). 
     Naturally occurring emulsifiers used in emulsion formulations include lanolin, beeswax, phosphatides, lecithin, and acacia. Absorption bases possess hydrophilic properties such that they can soak up water to form w/o emulsions yet retain their semisolid consistencies, such as anhydrous lanolin and hydrophilic petrolatum. Finely divided solids have also been used as good emulsifiers especially in combination with surfactants and in viscous preparations. These include polar inorganic solids, such as heavy metal hydroxides, nonswelling clays such as bentonite, attapulgite, hectorite, kaolin, montmorillonite, colloidal aluminum silicate and colloidal magnesium aluminum silicate, pigments, and nonpolar solids such as carbon or glyceryl tristearate. 
     A large variety of non-emulsifying materials are also included in emulsion formulations and contribute to the properties of emulsions. These include fats, oils, waxes, fatty acids, fatty alcohols, fatty esters, humectants, hydrophilic colloids, preservatives and antioxidants (Block, in Pharmaceutical Dosage Forms, Lieberman, Rieger and Banker (Eds.), 1988, Marcel Dekker, Inc., New York, N.Y., volume 1, p. 335; Idson, in Pharmaceutical Dosage Forms, Lieberman, Rieger and Banker (Eds.), 1988, Marcel Dekker, Inc., New York, N.Y., volume 1, p. 199). 
     Hydrophilic colloids or hydrocolloids include naturally occurring gums and synthetic polymers such as polysaccharides (for example, acacia, agar, alginic acid, carrageenan, guar gum, karaya gum, and tragacanth), cellulose derivatives (for example, carboxymethylcellulose and carboxypropylcellulose), and synthetic polymers (for example, carbomers, cellulose ethers, and carboxyvinyl polymers). These disperse or swell in water to form colloidal solutions that stabilize emulsions by forming strong interfacial films around the dispersed-phase droplets and by increasing the viscosity of the external phase. 
     Since emulsions often contain a number of ingredients such as carbohydrates, proteins, sterols, and phosphatides that can readily support the growth of microbes, these formulations often incorporate preservatives. Commonly used preservatives included in emulsion formulations include methyl paraben, propyl paraben, quaternary ammonium salts, benzalkonium chloride, esters of p-hydroxybenzoic acid, and boric acid. Antioxidants are also commonly added to emulsion formulations to prevent deterioration of the formulation. Antioxidants used can be free radical scavengers such as tocopherols, alkyl gallates, butylated hydroxyanisole, butylated hydroxytoluene, or reducing agents such as ascorbic acid and sodium metabisulfite, and antioxidant synergists such as citric acid, tartaric acid, and lecithin. 
     The application of emulsion formulations via dermatological, oral and parenteral routes and methods for their manufacture have been reviewed in the literature (see e.g., Ansel&#39;s Pharmaceutical Dosage Forms and Drug Delivery Systems, Allen, L V., Popovich N G., and Ansel H C., 2004, Lippincott Williams &amp; Wilkins (8th ed.), New York, N.Y.; Idson, in Pharmaceutical Dosage Forms, Lieberman, Rieger and Banker (Eds.), 1988, Marcel Dekker, Inc., New York, N.Y., volume 1, p. 199). Emulsion formulations for oral delivery have been very widely used because of ease of formulation, as well as efficacy from an absorption and bioavailability standpoint (see e.g., Ansel&#39;s Pharmaceutical Dosage Forms and Drug Delivery Systems, Allen, L V., Popovich N G., and Ansel H C., 2004, Lippincott Williams &amp; Wilkins (8th ed.), New York, N.Y.; Rosoff, in Pharmaceutical Dosage Forms, Lieberman, Rieger and Banker (Eds.), 1988, Marcel Dekker, Inc., New York, N.Y., volume 1, p. 245; Idson, in Pharmaceutical Dosage Forms, Lieberman, Rieger and Banker (Eds.), 1988, Marcel Dekker, Inc., New York, N.Y., volume 1, p. 199). Mineral-oil base laxatives, oil-soluble vitamins, and high fat nutritive preparations are among the materials that have commonly been administered orally as o/w emulsions. 
     ii. Microemulsions 
     In one embodiment of the present 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). Microemulsions commonly are prepared via a combination of three to five components that include oil, water, surfactant, cosurfactant and electrolyte. Whether the microemulsion is of the water-in-oil (w/o) or an oil-in-water (o/w) type is dependent on the properties of the oil and surfactant used and on the structure and geometric packing of the polar heads and hydrocarbon tails of the surfactant molecules (Schott, in Remington&#39;s Pharmaceutical Sciences, Mack Publishing Co., Easton, Pa., 1985, p. 271). 
     The phenomenological approach utilizing phase diagrams has been extensively studied and has yielded a comprehensive knowledge, to one skilled in the art, of how to formulate microemulsions (see e.g., Ansel&#39;s Pharmaceutical Dosage Forms and Drug Delivery Systems, Allen, L V., Popovich N G., and Ansel H C., 2004, Lippincott Williams &amp; Wilkins (8th ed.), New York, N.Y.; Rosoff, in Pharmaceutical Dosage Forms, Lieberman, Rieger and Banker (Eds.), 1988, Marcel Dekker, Inc., New York, N.Y., volume 1, p. 245; Block, in Pharmaceutical Dosage Forms, Lieberman, Rieger and Banker (Eds.), 1988, Marcel Dekker, Inc., New York, N.Y., volume 1, p. 335). Compared to conventional emulsions, microemulsions offer the advantage of solubilizing water-insoluble drugs in a formulation of thermodynamically stable droplets that are formed spontaneously. 
     Surfactants used in the preparation of microemulsions include, but are not limited to, ionic surfactants, non-ionic surfactants, Brij™ 96, polyoxyethylene oleyl ethers, polyglycerol fatty acid esters, tetraglycerol monolaurate (ML310), tetraglycerol monooleate (MO310), hexaglycerol monooleate (PO310), hexaglycerol pentaoleate (PO500), decaglycerol monocaprate (MCA750), decaglycerol monooleate (MO750), decaglycerol sequioleate (SO750), decaglycerol decaoleate (DAO750), alone or in combination with cosurfactants. The cosurfactant, usually a short-chain alcohol such as ethanol, 1-propanol, or 1-butanol, serves to increase the interfacial fluidity by penetrating into the surfactant film and consequently creating a disordered film because of the void space generated among surfactant molecules. Microemulsions can, however, be prepared without the use of cosurfactants and alcohol-free self-emulsifying microemulsion systems are known in the art. The aqueous phase can typically be, but is not limited to, water, an aqueous solution of the drug, glycerol, PEG300, PEG400, polyglycerols, propylene glycols, and derivatives of ethylene glycol. The oil phase can include, but is not limited to, materials such as Captex™ 300, Captex™ 355, Capmul™ MCM, fatty acid esters, medium chain (C8-C12) mono, di, and tri-glycerides, polyoxyethylated glyceryl fatty acid esters, fatty alcohols, polyglycolized glycerides, saturated polyglycolized C8-C10 glycerides, vegetable oils, and silicone oil. 
     Microemulsions are particularly of interest from the standpoint of drug solubilization and the enhanced absorption of drugs. Lipid based microemulsions (both o/w and w/o) have been proposed to enhance the oral bioavailability of drugs, including peptides (see e.g., U.S. Pat. Nos. 6,191,105; 7,063,860; 7,070,802; 7,157,099; Constantinides et al., Pharmaceutical Research, 1994, 11, 1385-1390; Ritschel, Meth. Find. Exp. Clin. Pharmacol., 1993, 13, 205). Microemulsions afford advantages of improved drug solubilization, protection of drug from enzymatic hydrolysis, possible enhancement of drug absorption due to surfactant-induced alterations in membrane fluidity and permeability, ease of preparation, ease of oral administration over solid dosage forms, improved clinical potency, and decreased toxicity (see e.g., U.S. Pat. Nos. 6,191,105; 7,063,860; 7,070,802; 7,157,099; Constantinides et al., Pharmaceutical Research, 1994, 11, 1385; Ho et al., J. Pharm. Sci., 1996, 85, 138-143). Often microemulsions can form spontaneously when their components are brought together at ambient temperature. This can be particularly advantageous when formulating thermolabile drugs, peptides or iRNAs. Microemulsions have also been effective in the transdermal delivery of active components in both cosmetic and pharmaceutical applications. It is expected that the microemulsion compositions and formulations of the present invention will facilitate the increased systemic absorption of iRNAs and nucleic acids from the gastrointestinal tract, as well as improve the local cellular uptake of iRNAs and nucleic acids. 
     Microemulsions of the present invention can also contain additional components and additives such as sorbitan monostearate (Grill 3), Labrasol, and penetration enhancers to improve the properties of the formulation and to enhance the absorption of the iRNAs and nucleic acids of the present invention. Penetration enhancers used in the microemulsions of the present invention can be classified as belonging to one of five broad categories-surfactants, fatty acids, bile salts, chelating agents, and non-chelating non-surfactants (Lee et al., Critical Reviews in Therapeutic Drug Carrier Systems, 1991, p. 92). Each of these classes has been discussed above. 
     iii. Microparticles 
     an RNAi agent of the 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). Such compounds are well known in the art. 
     v. Carriers 
     Certain compositions of the present invention also incorporate carrier compounds in the formulation. As used herein, “carrier compound” or “carrier” can refer to a nucleic acid, or analog thereof, which is inert (i.e., does not possess biological activity per se) but is recognized as a nucleic acid by in vivo processes that reduce the bioavailability of a nucleic acid having biological activity by, for example, degrading the biologically active nucleic acid or promoting its removal from circulation. The coadministration of a nucleic acid and a carrier compound, typically with an excess of the latter substance, can result in a substantial reduction of the amount of nucleic acid recovered in the liver, kidney, or other extracirculatory reservoirs, presumably due to competition between the carrier compound and the nucleic acid for a common receptor. For example, the recovery of a partially phosphorothioate dsRNA in hepatic tissue can be reduced when it is coadministered with polyinosinic acid, dextran sulfate, polycytidic acid or 4-acetamido-4′isothiocyano-stilbene-2,2′-disulfonic acid (Miyao et al., DsRNA Res. Dev., 1995, 5, 115-121; Takakura et al., DsRNA &amp; Nucl. Acid Drug Dev., 1996, 6, 177-183. 
     vi. Excipients 
     In contrast to a carrier compound, a “pharmaceutical carrier” or “excipient” is a pharmaceutically acceptable solvent, suspending agent or any other pharmacologically inert vehicle for delivering one or more nucleic acids to an animal. The excipient can be liquid or solid and is selected, with the planned manner of administration in mind, so as to provide for the desired bulk, consistency, etc., when combined with a nucleic acid and the other components of a given pharmaceutical composition. Typical pharmaceutical carriers include, but are not limited to, binding agents (e.g., pregelatinized maize starch, polyvinylpyrrolidone or hydroxypropyl methylcellulose, etc.); fillers (e.g., lactose and other sugars, microcrystalline cellulose, pectin, gelatin, calcium sulfate, ethyl cellulose, polyacrylates or calcium hydrogen phosphate, etc.); lubricants (e.g., magnesium stearate, talc, silica, colloidal silicon dioxide, stearic acid, metallic stearates, hydrogenated vegetable oils, corn starch, polyethylene glycols, sodium benzoate, sodium acetate, etc.); disintegrants (e.g., starch, sodium starch glycolate, etc.); and wetting agents (e.g., sodium lauryl sulphate, etc). 
     Pharmaceutically acceptable organic or inorganic excipients suitable for non-parenteral administration which do not deleteriously react with nucleic acids can also be used to formulate the compositions of the present invention. Suitable pharmaceutically acceptable carriers include, but are not limited to, water, salt solutions, alcohols, polyethylene glycols, gelatin, lactose, amylose, magnesium stearate, talc, silicic acid, viscous paraffin, hydroxymethylcellulose, polyvinylpyrrolidone; and the like. 
     Formulations for topical administration of nucleic acids can include sterile and non-sterile aqueous solutions, non-aqueous solutions in common solvents such as alcohols, or solutions of the nucleic acids in liquid or solid oil bases. The solutions can also contain buffers, diluents, and other suitable additives. Pharmaceutically acceptable organic or inorganic excipients suitable for non-parenteral administration which do not deleteriously react with nucleic acids can be used. 
     Suitable pharmaceutically acceptable excipients include, but are not limited to, water, salt solutions, alcohol, polyethylene glycols, gelatin, lactose, amylose, magnesium stearate, talc, silicic acid, viscous paraffin, hydroxymethylcellulose, polyvinylpyrrolidone, and the like. 
     vii. Other Components 
     The compositions of the present 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, and/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, and/or dextran. The suspension can also contain stabilizers. 
     In some embodiments, pharmaceutical compositions featured in the invention include (a) one or more iRNA compounds and (b) one or more agents which function by a non-RNAi mechanism and which are useful in treating a GCK-associated disorder. Toxicity and therapeutic efficacy of such compounds can be determined by standard pharmaceutical procedures in cell cultures or experimental animals, e.g., for determining the LD 50  (the dose lethal to 50% of the population) and the ED 50  (the dose therapeutically effective in 50% of the population). The dose ratio between toxic and therapeutic effects is the therapeutic index and it can be expressed as the ratio LD 50 /ED 50 . Compounds that exhibit high therapeutic indices are preferred. 
     The data obtained from cell culture assays and animal studies can be used in formulating a range of dosage for use in humans. The dosage of compositions featured herein in the invention lies generally within a range of circulating concentrations that include the ED 50  with little or no toxicity. The dosage can vary within this range depending upon the dosage form employed and the route of administration utilized. For any compound used in the methods featured in the invention, the therapeutically effective dose can be estimated initially from cell culture assays. A dose can be formulated in animal models to achieve a circulating plasma concentration range of the compound or, when appropriate, of the polypeptide product of a target sequence (e.g., achieving a decreased concentration of the polypeptide) that includes the IC 50  (i.e., the concentration of the test compound which achieves a half-maximal inhibition of symptoms) as determined in cell culture. Such information can be used to more accurately determine useful doses in humans. Levels in plasma can be measured, for example, by high performance liquid chromatography. 
     In addition to their administration, as discussed above, the iRNAs featured in the invention can be administered in combination with other known agents effective in treatment of pathological processes mediated by GCK expression. 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 of the Invention 
     The present invention also provides methods of using an iRNA of the invention and/or a composition containing an iRNA of the invention to reduce and/or inhibit glucokinase (GCK) expression in a cell. The methods include contacting the cell with a dsRNA of the invention and maintaining the cell for a time sufficient to obtain degradation of the mRNA transcript of a GCK gene, thereby inhibiting expression of the GCK gene in the cell. Reduction in gene expression can be assessed by any methods known in the art. For example, a reduction in the expression of GCK may be determined by determining the mRNA expression level of GCK using methods routine to one of ordinary skill in the art, e.g., northern blotting, qRT-PCR; by determining the protein level of GCK using methods routine to one of ordinary skill in the art, such as western blotting, immunological techniques. A reduction in the expression of GCK may also be assessed indirectly by measuring a decrease in biological activity of GCK. 
     In the methods of the invention the cell may be contacted in vitro or in vivo, i.e., the cell may be within a subject. 
     A cell suitable for treatment using the methods of the invention may be any cell that expresses a GCK gene. 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 or a non-human primate cell, e.g., a monkey cell or a chimpanzee cell), a non-primate cell (such as a cow cell, a pig cell, a camel cell, a llama cell, a horse cell, a goat cell, a rabbit cell, a sheep cell, a hamster, a guinea pig cell, a cat cell, a dog cell, a rat cell, a mouse cell, a lion cell, a tiger cell, a bear cell, or a buffalo cell), a bird cell (e.g., a duck cell or a goose cell), or a whale cell. In one embodiment, the cell is a human cell, e.g., a human liver cell. 
     GCK expression is inhibited in the cell by at least about 5, 6, 7, 8, 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, 36, 37, 38, 39, 40, 41, 42, 43, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, or about 100%. In preferred embodiments, GCK expression is inhibited by at least 20%. 
     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 GCK gene of the mammal to be treated. When the organism to be treated is a mammal such as a human, the composition can be administered by any means known in the art including, but not limited to oral, intraperitoneal, or parenteral routes, including intracranial (e.g., intraventricular, intraparenchymal, and intrathecal), intravenous, intramuscular, subcutaneous, transdermal, airway (aerosol), nasal, rectal, and topical (including buccal and sublingual) administration. In certain embodiments, the compositions are administered by intravenous infusion or injection. In certain embodiments, the compositions are administered by subcutaneous injection. 
     In some embodiments, the administration is via a depot injection. A depot injection may release the iRNA in a consistent way over a prolonged time period. Thus, a depot injection may reduce the frequency of dosing needed to obtain a desired effect, e.g., a desired inhibition of GCK, or a therapeutic or prophylactic effect. A depot injection may also provide more consistent serum concentrations. Depot injections may include subcutaneous injections or intramuscular injections. In preferred embodiments, the depot injection is a subcutaneous injection. 
     In some embodiments, the administration is via a pump. The pump may be an external pump or a surgically implanted pump. In certain embodiments, the pump is a subcutaneously implanted osmotic pump. In other embodiments, the pump is an infusion pump. An infusion pump may be used for intravenous, subcutaneous, arterial, or epidural infusions. In preferred embodiments, the infusion pump is a subcutaneous infusion pump. In other embodiments, the pump is a surgically implanted pump that delivers the iRNA to the liver. 
     The mode of administration may be chosen based upon whether local or systemic treatment is desired and based upon the area to be treated. The route and site of administration may be chosen to enhance targeting. 
     In one aspect, the present invention also provides methods for inhibiting the expression of a GCK gene in a mammal. The methods include administering to the mammal a composition comprising a dsRNA that targets a GCK gene in a cell of the mammal and maintaining the mammal for a time sufficient to obtain degradation of the mRNA transcript of the GCK gene, thereby inhibiting expression of the GCK gene in the cell. Reduction in gene expression can be assessed by any methods known it the art and by methods, e.g. qRT-PCR, described herein. Reduction in protein production can be assessed by any methods known it the art and by methods, e.g. ELISA, described herein. In one embodiment, a puncture liver biopsy sample serves as the tissue material for monitoring the reduction in GCK gene and/or protein expression. 
     The present invention further provides methods of treatment of 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 and/or inhibition of GCK expression, in a therapeutically effective amount of an iRNA targeting a GCK gene or a pharmaceutical composition comprising an iRNA targeting a GCK gene. 
     In addition, the present invention provides methods of increasing the blood glucose levels, e.g., fasting blood glucose levels, in a subject having a disease or disorder that would benefit from a reduction in GCK expression. The methods include administering an iRNA of the invention to the subject in a therapeutically effective amount of an iRNA targeting a GCK gene or a pharmaceutical composition comprising an iRNA targeting a GCK gene. 
     Further, the present invention provides methods of decreasing plasma lactate levels in a subject having a disease or disorder that would benefit from a reduction in GCK expression, e.g., a GSD, e.g., type Ia GSD. The methods include administering an iRNA of the invention to the subject in a therapeutically effective amount of an iRNA targeting a GCK gene or a pharmaceutical composition comprising an iRNA targeting a GCK gene. 
     The present invention also provides methods of decreasing plasma uric acid levels in a subject having a disease or disorder that would benefit from a reduction in GCK expression, e.g., a GSD, e.g., type Ia GSD. The methods include administering an iRNA of the invention to the subject in a therapeutically effective amount of an iRNA targeting a GCK gene or a pharmaceutical composition comprising an iRNA targeting a GCK gene. 
     The present invention provides methods of decreasing plasma triglyceride levels in a subject having a disease or disorder that would benefit from a reduction in GCK expression, e.g., a GSD, e.g., type Ia GSD. The methods include administering an iRNA of the invention to the subject in a therapeutically effective amount of an iRNA targeting a GCK gene or a pharmaceutical composition comprising an iRNA targeting a GCK gene. 
     The present invention further provides methods of decreasing total plasma cholesterol levels in a subject having a disease or disorder that would benefit from a reduction in GCK expression, e.g., a GSD, e.g., type Ia GSD. The methods include administering an iRNA of the invention to the subject in a therapeutically effective amount of an iRNA targeting a GCK gene or a pharmaceutical composition comprising an iRNA targeting a GCK gene. 
     The present invention also provides methods of decreasing hepatomegaly in a subject having a disease or disorder that would benefit from a reduction in GCK expression, e.g., a GSD, e.g., type Ia GSD. The methods include administering an iRNA of the invention to the subject in a therapeutically effective amount of an iRNA targeting a GCK gene or a pharmaceutical composition comprising an iRNA targeting a GCK 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 a reduction and/or inhibition of GCK gene expression include those having a glycogen storage disease (GSD), e.g., type Ia GSD. In one embodiment, subjects that would benefit from a reduction and/or inhibition of GCK gene expression are those having one or more signs or symptoms associated with a glycogen storage disease (GSD), e.g., type Ia GSD, including, but not limited to, hypoglycemia, lactic acidosis, hyperuricemia, hyperlipidemia, hepatomegaly, kidney disease as a result of glycogen accumulation, hunger, jitteriness, lethargy, apnea, seizures, diaphoresis, confusion, headaches, dizziness, unusual mood or behavior changes, loss of consciousness, coma, muscle cramps, bleeding diathesis, short stauture, osteoporosis, delayed puberty, gout, renal disease, systemic hypertension, pulmonary hypertension, hepatic adenomas, pancreatitis, anemia, vitamin D deficiency, polycystic ovaries, irregular menstrual cycles, menorrhagia, and eruptive xanthomata. 
     Treatment of a subject that would benefit from a reduction and/or inhibition of GCK gene expression and normalization of blood glucose levels includes therapeutic treatment (e.g., of a subject suffering from a glycogen storage disease (GSD)) and prophylactic treatment (e.g., of a subject that does not meet the diagnostic criteria of a glycogen storage disease (GSD), or a subject who may be at risk of developing a glycogen storage disease (GSD)). 
     The invention further provides methods for the use of an iRNA or a pharmaceutical composition thereof, e.g., for treating a subject that would benefit from reduction and/or inhibition of GCK expression, e.g., a subject having a glycogen storage disease (GSD), e.g., type Ia GSD, in combination with other pharmaceuticals and/or other therapeutic methods, e.g., with known pharmaceuticals and/or known therapeutic methods, such as, for example, those which are currently employed for treating these disorders. For example, in certain embodiments, an iRNA targeting GCK is administered to a subject having a GSD in combination with, e.g., antihypertensive agents, such as Diazoxide, somatostatin analogues, such as Octreotide, calcium channel blockers, such as, Nifedipine, Thiazide diuretics, such as chlorothiazide (e.g., in combination with Diazoxide), intravenous infusion of glucagon, parenterally adminsitered dextrose; and/or partial pancreatectomy. In some embodiments, an iRNA targeting GCK is administered to a subject having type Ia GSD in combination with, e.g., a sodium-glucose co-transporter 2 (SGLT2) inhibitor, e.g., Dapagliflozin, Canagliflozin, Ipragliflozin (ASP-1941), Tofogliflozin, Empagliflozin, Sergliflozin etabonate, Remogliflozin etabonate (BHVO91009), and Ertugliflozin (PF-04971729/MK-8835). 
     The iRNA and additional therapeutic agents may be administered at the same time and/or in the same combination, e.g., parenterally, or the additional therapeutic agent can be administered as part of a separate composition or at separate times and/or by another method known in the art or described herein. 
     In one embodiment, the method includes administering a composition featured herein such that expression of the target GCK gene is decreased, such as for about 1, 2, 3, 4, 5, 6, 7, 8, 12, 16, 18, 24 hours, 28, 32, or about 36 hours. In one embodiment, expression of the target GCK gene is decreased for an extended duration, e.g., at least about two, three, four days or more, e.g., about one week, two weeks, three weeks, or four weeks or longer. 
     Preferably, the iRNAs useful for the methods and compositions featured herein specifically target RNAs (primary or processed) of the target GCK gene. Compositions and methods for inhibiting the expression of these genes using iRNAs can be prepared and performed as described herein. 
     Administration of the dsRNA according to the methods of the invention may result in a reduction of the severity, signs, symptoms, and/or markers of such diseases or disorders in a patient with a glycogen storage disease (GSD), e.g., type Ia GSD. By “reduction” in this context is meant a statistically significant decrease in such level. The reduction can be, for example, at least about 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100%. 
     Efficacy of treatment or prevention of disease can be assessed, for example by measuring disease progression, disease remission, symptom severity, reduction in pain, quality of life, dose of a medication required to sustain a treatment effect, level of a disease marker, or any other measurable parameter appropriate for a given disease being treated or targeted for prevention. It is well within the ability of one skilled in the art to monitor efficacy of treatment or prevention by measuring any one of such parameters, or any combination of parameters. For example, efficacy of treatment of a glycogen storage disease (GSD) may be assessed, for example, by periodic monitoring of, e.g., blood glucose levels. Comparisons of the later readings with the initial readings provide a physician an indication of whether the treatment is effective. It is well within the ability of one skilled in the art to monitor efficacy of treatment or prevention by measuring any one of such parameters, or any combination of parameters. In connection with the administration of an iRNA targeting GCK or pharmaceutical composition thereof, “effective against” a glycogen storage disease (GSD) indicates that administration in a clinically appropriate manner results in a beneficial effect for at least a statistically significant fraction of patients, such as a improvement of symptoms, a cure, a reduction in disease, extension of life, improvement in quality of life, or other effect generally recognized as positive by medical doctors familiar with treating a glycogen storage disease (GSD) and the related causes and effects. 
     A treatment or preventive effect is evident when there is a statistically significant improvement in one or more parameters of disease status, or by a failure to worsen or to develop symptoms where they would otherwise be anticipated. As an example, a favorable change of at least 10% in a measurable parameter of disease, and preferably at least 20%, 30%, 40%, 50% or more can be indicative of effective treatment. Efficacy for a given iRNA drug or formulation of that drug can also be judged using an experimental animal model for the given disease as known in the art. When using an experimental animal model, efficacy of treatment is evidenced when a statistically significant reduction in a marker or symptom is observed. 
     Alternatively, the efficacy can be measured by a reduction in the severity of disease as determined by one skilled in the art of diagnosis based on a clinically accepted disease severity grading scale such as those provided above. Any positive change resulting in e.g., lessening of severity of disease measured using the appropriate scale, represents adequate treatment using an iRNA or iRNA formulation as described herein. 
     Subjects can be administered a therapeutic amount of dsRNA, such as about 0.01 mg/kg to about 200 mg/kg. 
     The iRNA can be administered by intravenous infusion over a period of time, on a regular basis. In certain embodiments, after an initial treatment regimen, the treatments can be administered on a less frequent basis. Administration of the iRNA can reduce GCK levels, e.g., in a cell, tissue, blood, urine, or other compartment of the patient by at least about 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 39, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99% or more. 
     Before administration of a full dose of the iRNA, patients can be administered a smaller dose, such as a 5% infusion reaction, and monitored for adverse effects, such as an allergic reaction. In another example, the patient can be monitored for unwanted immunostimulatory effects, such as increased cytokine (e.g., TNF-alpha or INF-alpha) levels. 
     Alternatively, the iRNA can be administered subcutaneously, i.e., by subcutaneous injection. One or more injections may be used to deliver the desired daily 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 every other day or to once a year. In certain embodiments, the iRNA is administered about once per month to about once per quarter (i.e., about once every three months). 
     Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. Although methods and materials similar or equivalent to those described herein can be used in the practice or testing of the iRNAs and methods featured in the invention, suitable methods and materials are described below. All publications, patent applications, patents, and other references mentioned herein are incorporated by reference in their entirety. In case of conflict, the present specification, including definitions, will control. In addition, the materials, methods, and examples are illustrative only and not intended to be limiting. 
     EXAMPLES 
     Example 1. iRNA Design, Synthesis, Selection, and In Vitro Evaluation 
     This Example describes methods for the design, synthesis, selection, and in vitro evaluation of GCK iRNA agents. 
     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. 
     Bioinformatics 
     A set of siRNAs targeting the human GCK, “glucokinase (hexokinase 4)”, (human: NCBI refseqID NM_033507; NCBI GenelD: 2645), as well as toxicology-species GCK orthologs (cynomolgus monkey: XM_005549685; mouse: NM_010292; rat, NM_012565) were designed using custom R and Python scripts. The human NM_033507 REFSEQ mRNA, version 1, has a length of 2442 bases. The rationale and method for the set of siRNA designs is as follows: the predicted efficacy for every potential 19mer iRNA from position 1 through position 2442 (the coding region and 3′ UTR) was determined with a linear model derived the direct measure of mRNA knockdown from more than 20,000 distinct iRNA designs targeting a large number of vertebrate genes. Subsets of the GCK iRNAs were designed with perfect or near-perfect matches between human, cynomolgus and rhesus monkey. A further subset was designed with perfect or near-perfect matches to mouse and rat GCK orthologs. For each strand of the iRNA, a custom Python script was used in a brute force search to measure the number and positions of mismatches between the iRNA 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 iRNA, 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 assuming the value of each weighted mismatch. Preference was given to iRNAs whose antisense score in human and cynomolgus monkey was &gt;=2.0 and predicted efficacy was &gt;=50% knockdown of the GCK transcript. 
     Synthesis of GCK Sequences 
     Synthesis of GCK Single Strands and Duplexes 
     GCK siRNA sequences were synthesized at 1 umol scale on 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, RNA, DNA and other modified nucleosides were introduced in the sequences 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, single strands were cleaved from the solid support and deprotected in sealed 96 deep well plates using 200 μL Aqueous Methylamine reagent at 60° C. for 20 minutes. For sequences containing 2′ ribo residues (2′-OH) that are protected with 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 μL of dimethyl sulfoxide (DMSO) and 300 μl 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 and the supernatant 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 GCK 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 uM in 1×PBS and then submitted for in vitro screening assays. 
     Cell Culture and Transfections for Single Dose and Dose Response Studies 
     Primary mouse hepatocytes (PMH) (GIBCO) or Primary Cynomolgus monkey hepatocytes (PCH) (Celsis) were transfected by adding 4.9 μl of Opti-MEM plus 0.1 μl of Lipofectamine RNAiMax per well (Invitrogen, Carlsbad Calif. cat #13778-150) to 5 μl of siRNA duplexes per well into a 384-well plate and incubated at room temperature for 15 minutes. Forty μl of William&#39;s E Medium (Life Tech) containing about 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 and 0.1 nM final duplex concentration. 
     Total RNA Isolation Using DYNABEADS mRNA Isolation Kit (Invitrogen, Part #: 610-12) 
     Total RNA was isolated using an automated protocol on a BioTek-EL406 platform using DYNABEADs (Invitrogen, cat #61012). Briefly, 50 μl of Lysis/Binding Buffer and 25 μl of lysis buffer containing 3 μl 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 μl Wash Buffer A and once with Wash Buffer B. Beads were then washed with 150 μl Elution Buffer, re-captured and supernatant removed. 
     cDNA Synthesis Using ABI High Capacity cDNA Reverse Transcription Kit (Applied Biosystems, Foster City, Calif., Cat #4368813) 
     Ten μl of a master mix containing 1 μl 10× Buffer, 0.4 μl 125× dNTPs, 1 μl 10× Random primers, 0.5 μl Reverse Transcriptase, 0.5 μl RNase inhibitor and 6.6 μl of H 2 O per reaction was added to the RNA isolated as described above. Plates were sealed, mixed, and incubated on an electromagnetic shaker for 10 minutes at room temperature, followed by 2 h 37° C. Plates were then incubated at 80° C. for 8 minutes. 
     Real Time PCR 
     Two μl of cDNA was added to a master mix containing 0.5 μl GAPDH TaqMan Probe (Applied Biosystems Cat #4326317E), or 0.5 μl of Custom made Cyno GAPDH Taqman Probe, 0.5 μl GCK mouse probe (Mm00439129 ml) or 0.5 μl cyno probe (Mf02827184 ml) and 5 μl Lightcycler 480 probe master mix (Roche Cat #04887301001) per well in a 384 well plate (Roche cat #04887301001). Real time PCR is done in an Roche Lightcycler Real Time PCR system (Roche) using the ΔΔCt(RQ) assay. Each duplex was tested in two independent transfections. 
     To calculate relative fold change, real time data are analyzed using the ΔΔCt method and normalized to assays performed with cells transfected with a non-targeting control siRNA, AD-1955 
     The sense and antisense sequences of AD-1955 are: 
     
       
         
           
               
               
            
               
                   
                 SENSE: 
               
               
                   
                 (SEQ ID NO: 29) 
               
               
                   
                 cuuAcGcuGAGuAcuucGAdTsdT 
               
               
                   
                   
               
               
                   
                 ANTISENSE: 
               
               
                   
                 (SEQ ID NO: 30) 
               
               
                   
                 UCGAAGuACUcAGCGuAAGdTsdT. 
               
            
           
         
       
     
     
       
         
           
               
             
               
                 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 
               
               
                 Af 
                 2′-fluoroadenosine-3′-phosphate 
               
               
                 Afs 
                 2′-fluoroadenosine-3′-phosphorothioate 
               
               
                 As 
                 adenosine-3′-phosphorothioate 
               
               
                 C 
                 cytidine-3′-phosphate 
               
               
                 Cf 
                 2′-fluorocytidine-3′-phosphate 
               
               
                 Cfs 
                 2′-fluorocytidine-3′-phosphorothioate 
               
               
                 Cs 
                 cytidine-3′-phosphorothioate 
               
               
                 G 
                 guanosine-3′-phosphate 
               
               
                 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 (G, A, C, T or U) 
               
               
                 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 
               
               
                 dT 
                 2′-deoxythymidine-3′-phosphate 
               
               
                 dC 
                 2′-deoxycytidine-3′-phosphate 
               
               
                   
               
            
           
         
       
     
     A detailed list of the unmodified GCK sense and antisense strand sequences is shown in Table 2 and a detailed list of the modified GCK sense and antisense strand sequences is shown in Table 3. 
     Table 4 shows the results of a single dose screen in primary mouse hepatocytes transfected with the indicated modified siRNAs. Data are expressed as percent of message remaining relative to cells treated with a non-targeting control siRNA. 
     Table 5 shows the results of a single dose screen in primary  Cynomologous  hepatocytes transfected with the indicated modified siRNAs. Data are expressed as percent of message remaining relative to cells treated with a non-targeting control siRNA. 
     
       
         
           
               
             
               
                 TABLE 2 
               
             
            
               
                   
               
               
                 GCK Unmodified Sequences 
               
            
           
           
               
               
               
               
               
               
               
               
               
               
            
               
                   
                   
                 Sense 
                 SEQ 
                   
                 Anti- 
                 Antisense 
                 SEQ 
                   
                   
               
               
                 Duplex 
                 Sense 
                 Sequence 
                 ID 
                 Postion in 
                 sense 
                 Sequence 
                 ID 
                 Postion in 
                   
               
               
                 ID 
                 ID 
                 (5′-3′) 
                 NO: 
                 NM_033507.1 
                 ID 
                 (5′-3′) 
                 NO: 
                 NM_033507.1 
                 cross_ sp 
               
               
                   
               
            
           
           
               
               
               
               
               
               
               
               
               
               
            
               
                 AD-69366 
                 A-139669 
                 GAGGACCUGAA 
                 31 
                 _253-273_s 
                 A-139670 
                 UAUCACCUUCUU 
                 120 
                 _251-273_as 
                 hcmr 
               
               
                   
                   
                 GAAGGUGAUA 
                   
                   
                   
                 CAGGUCCUCCU 
                   
                   
                   
               
               
                   
               
               
                 AD-69368 
                 A-139673 
                 GGACCUGAAGA 
                 32 
                 _255-275_s 
                 A-139674 
                 UUCAUCACCUUC 
                 121 
                 _253-275_as 
                 hcmr 
               
               
                   
                   
                 AGGUGAUGAA 
                   
                   
                   
                 UUCAGCUCCUC 
                   
                   
                   
               
               
                   
               
               
                 AD-69411 
                 A-139758 
                 GGACCUGAAGA 
                 33 
                 _257-277_s 
                 A-139759 
                 UAUCACCUUCUU 
                 122 
                 _255-277_as 
                 hcmr 
               
               
                   
                   
                 AGGUGAUA 
                   
                   
                   
                 CAGGUCC 
                   
                   
                   
               
               
                   
               
               
                 AD-69413 
                 A-139762 
                 ACCUGAAGAAG 
                 34 
                 _259-279_s 
                 A-139763 
                 UUCAUCACCUUC 
                 123 
                 _257-279_as 
                 hcmr 
               
               
                   
                   
                 GUGAUGAA 
                   
                   
                   
                 UUCAGGU 
                   
                   
                   
               
               
                   
               
               
                 AD-69367 
                 A-139671 
                 GGUGAUGAGAC 
                 35 
                 _267-287_s 
                 A-139672 
                 UUCUGCAUCCGU 
                 124 
                 _265-287_as 
                 hcmr 
               
               
                   
                   
                 GGAUGCAGAA 
                   
                   
                   
                 CUCAUCACCUU 
                   
                   
                   
               
               
                   
               
               
                 AD-69369 
                 A-139675 
                 UGAUGAGACGG 
                 36 
                 _269-289_s 
                 A-139676 
                 UCUUCUGCAUCC 
                 125 
                 _267-289_as 
                 hcmr 
               
               
                   
                   
                 AUGCAGAAGA 
                   
                   
                   
                 GUCUCAUCACC 
                   
                   
                   
               
               
                   
               
               
                 AD-69412 
                 A-139760 
                 UGAUGAGACGG 
                 37 
                 _271-291_s 
                 A-139761 
                 UUCUGCAUCCGU 
                 126 
                 _269-291_as 
                 hcmr 
               
               
                   
                   
                 AUGCAGAA 
                   
                   
                   
                 CUCAUCA 
                   
                   
                   
               
               
                   
               
               
                 AD-69414 
                 A-139764 
                 AUGAGACGGAU 
                 38 
                 _273-293_s 
                 A-139765 
                 UCUUCUGCAUCC 
                 127 
                 _271-293_as 
                 hcmr 
               
               
                   
                   
                 GCAGAAGA 
                   
                   
                   
                 GUCUCAU 
                   
                   
                   
               
               
                   
               
               
                 AD-69371 
                 A-139679 
                 ACGGAUGCAGA 
                 39 
                 _276-296_s 
                 A-139680 
                 UCCAUCUCCUUC 
                 128 
                 _274-296_as 
                 hcmr 
               
               
                   
                   
                 AGGAGAUGGA 
                   
                   
                   
                 UGCAUCCGUCU 
                   
                   
                   
               
               
                   
               
               
                 AD-69416 
                 A-139768 
                 GGAUGCAGAAG 
                 40 
                 _280-300_s 
                 A-139769 
                 UCCAUCUCCUUC 
                 129 
                 _278-300_as 
                 hcmr 
               
               
                   
                   
                 GAGAUGGA 
                   
                   
                   
                 UGCAUCC 
                   
                   
                   
               
               
                   
               
               
                 AD-69370 
                 A-139677 
                 ACCCAUGAAGA 
                 41 
                 _316-336_s 
                 A-139678 
                 UACACUGGCCUC 
                 130 
                 _314-336_as 
                 hcmr 
               
               
                   
                   
                 GGCCAGUGUA 
                   
                   
                   
                 UUCAUGGGUCU 
                   
                   
                   
               
               
                   
               
               
                 AD-69415 
                 A-139766 
                 CCAUGAAGAGG 
                 42 
                 _320-340_s 
                 A-139767 
                 UACACUGGCCUC 
                 131 
                 _318-340_as 
                 hcmr 
               
               
                   
                   
                 CCAGUGUA 
                   
                   
                   
                 UUCAUGG 
                   
                   
                   
               
               
                   
               
               
                 AD-69372 
                 A-139681 
                 CAGUGUGAAGA 
                 43 
                 _330-350_s 
                 A-139682 
                 UUGGGCAGCAUC 
                 132 
                 _328-350_as 
                 hc 
               
               
                   
                   
                 UGCUGCCCAA 
                   
                   
                   
                 UUCACACUGGC 
                   
                   
                   
               
               
                   
               
               
                 AD-69417 
                 A-139770 
                 GUGUGAAGAUG 
                 44 
                 _334-354_s 
                 A-139771 
                 UUGGGCAGCAUC 
                 133 
                 _332-354_as 
                 hc 
               
               
                   
                   
                 CUGCCCAA 
                   
                   
                   
                 UUCACAC 
                   
                   
                   
               
               
                   
               
               
                 AD-69373 
                 A-139683 
                 GUGAAGGUGGG 
                 45 
                 _436-456_s 
                 A-139684 
                 UUCACCUUCUCC 
                 134 
                 _434-456_as 
                 hc 
               
               
                   
                   
                 AGAAGGUGAA 
                   
                   
                   
                 CACCUUCACCA 
                   
                   
                   
               
               
                   
               
               
                 AD-69418 
                 A-139772 
                 GAAGGUGGGAG 
                 46 
                 _440-460_s 
                 A-139773 
                 UUCACCUUCUCC 
                 135 
                 _438-460_as 
                 hc 
               
               
                   
                   
                 AAGGUGAA 
                   
                   
                   
                 CACCUUC 
                   
                   
                   
               
               
                   
               
               
                 AD-69374 
                 A-139685 
                 GAGCGUGAAGA 
                 47 
                 _168-488_s 
                 A-139686 
                 UGGUGUUUGGUC 
                 136 
                 _466-488_as 
                 hc 
               
               
                   
                   
                 CCAAACACCA 
                   
                   
                   
                 UUCACGCUCCA 
                   
                   
                   
               
               
                   
               
               
                 AD-69419 
                 A-139774 
                 GCGUGAAGACC 
                 48 
                 _472-492_s 
                 A-139775 
                 UGGUGUUUGGUC 
                 137 
                 _470-492_as 
                 hc 
               
               
                   
                   
                 AAACACCA 
                   
                   
                   
                 UUCACGC 
                   
                   
                   
               
               
                   
               
               
                 AD-69375 
                 A-139687 
                 GAAGACCAAAC 
                 49 
                 _474-494_s 
                 A-139688 
                 UACAUCUGGUGU 
                 138 
                 _472-494_as 
                 hcmr 
               
               
                   
                   
                 ACCAGAUGUA 
                   
                   
                   
                 UUGGUCUUCAC 
                   
                   
                   
               
               
                   
               
               
                 AD-69420 
                 A-139776 
                 AGACCAAACAC 
                 50 
                 _478-498_s 
                 A-139777 
                 UACAUCUGGUGU 
                 139 
                 _476-498_as 
                 hcmr 
               
               
                   
                   
                 CAGAUGUA 
                   
                   
                   
                 UUGGUCU 
                   
                   
                   
               
               
                   
               
               
                 AD-69376 
                 A-139689 
                 GACUUCCUGGA 
                 51 
                 _565-585_s 
                 A-139690 
                 UUGAUGCUUGUC 
                 140 
                 _563-585_as 
                 hcmr 
               
               
                   
                   
                 CAAGCAUCAA 
                   
                   
                   
                 CAGGAAGUCGG 
                   
                   
                   
               
               
                   
               
               
                 AD-69377 
                 A-139691 
                 CUUCCUGGACA 
                 52 
                 _567-587_s 
                 A-139692 
                 AUCUGAUGCUUG 
                 141 
                 _565-587_as 
                 hcmr 
               
               
                   
                   
                 AGCAUCAGAU 
                   
                   
                   
                 UCCAGGAAGUC 
                   
                   
                   
               
               
                   
               
               
                 AD-69421 
                 A-139778 
                 CUUCCUGGACA 
                 53 
                 _569-589_s 
                 A-139779 
                 UUGAUGCUUGUC 
                 142 
                 _567-589_as 
                 hcmr 
               
               
                   
                   
                 AGCAUCAA 
                   
                   
                   
                 CAGGAAG 
                   
                   
                   
               
               
                   
               
               
                 AD-69422 
                 A-139780 
                 UCCUGGACAAG 
                 54 
                 _571-591_s 
                 A-139781 
                 AUCUGAUGCUUG 
                 143 
                 _569-591_as 
                 hcmr 
               
               
                   
                   
                 CAUCAGAU 
                   
                   
                   
                 UCCAGGA 
                   
                   
                   
               
               
                   
               
               
                 AD-69378 
                 A-139693 
                 CCUGGACAAGC 
                 55 
                 _570-590_s 
                 A-139694 
                 UUCAUCUGAUGC 
                 144 
                 _568-590_as 
                 hcmr 
               
               
                   
                   
                 AUCAGAUGAA 
                   
                   
                   
                 UUGUCCAGGAA 
                   
                   
                   
               
               
                   
               
               
                 AD-69379 
                 A-139695 
                 CUGGACAAGCA 
                 56 
                 _571-591_s 
                 A-139696 
                 UUUCAUCUGAUG 
                 145 
                 _569-591_as 
                 hcmr 
               
               
                   
                   
                 UCAGAUGAAA 
                   
                   
                   
                 CUUGUCCAGGA 
                   
                   
                   
               
               
                   
               
               
                 AD-69423 
                 A-139782 
                 UGGACAAGCAU 
                 57 
                 _574-594_s 
                 A-139783 
                 UUCAUCUGAUGC 
                 146 
                 _572-594_as 
                 hcmr 
               
               
                   
                   
                 CAGAUGAA 
                   
                   
                   
                 UUGUCCA 
                   
                   
                   
               
               
                   
               
               
                 AD-69424 
                 A-139784 
                 GGACAAGCAUC 
                 58 
                 _575-595_s 
                 A-139785 
                 UUUCAUCUGAUG 
                 147 
                 _573-595_as 
                 hcmr 
               
               
                   
                   
                 AGAUGAAA 
                   
                   
                   
                 CUUGUCC 
                   
                   
                   
               
               
                   
               
               
                 AD-69381 
                 A-139699 
                 AGGCACGAAGA 
                 59 
                 _634-654_s 
                 A-139700 
                 UUUAUCGAUGUC 
                 148 
                 _632-654_as 
                 hc 
               
               
                   
                   
                 CAUCGAUAAA 
                   
                   
                   
                 UUCGUGCCUCA 
                   
                   
                   
               
               
                   
               
               
                 AD-69426 
                 A-139788 
                 GCACGAAGACA 
                 60 
                 _638-658_s 
                 A-139789 
                 UUUAUCGAUGUC 
                 149 
                 _636-658_as 
                 hc 
               
               
                   
                   
                 UCGAUAAA 
                   
                   
                   
                 UUCGUGC 
                   
                   
                   
               
               
                   
               
               
                 AD-69382 
                 A-139701 
                 ACAUCGAUAAG 
                 61 
                 _644-664_s 
                 A-139702 
                 UAAGGAUGCCCU 
                 150 
                 _642-664_as 
                 hc 
               
               
                   
                   
                 GGCAUCCUUA 
                   
                   
                   
                 UAUCGAUGUCU 
                   
                   
                   
               
               
                   
               
               
                 AD-69427 
                 A-139790 
                 AUCGAUAAGGG 
                 62 
                 _648-668_s 
                 A-139791 
                 UAAGGAUGCCCU 
                 151 
                 _646-668_as 
                 hc 
               
               
                   
                   
                 CAUCCUUA 
                   
                   
                   
                 UAUCGAU 
                   
                   
                   
               
               
                   
               
               
                 AD-69383 
                 A-139703 
                 CUGGACCAAGG 
                 63 
                 _669-689_s 
                 A-139704 
                 UCCUUGAAGCCC 
                 152 
                 _667-6X9_as 
                 hcmr 
               
               
                   
                   
                 GCUUCAAGGA 
                   
                   
                   
                 UUGGUCCAGUU 
                   
                   
                   
               
               
                   
               
               
                 AD-69428 
                 A-139792 
                 GGACCAAGGGC 
                 64 
                 _673-693_s 
                 A-139793 
                 UCCUUGAAGCCC 
                 153 
                 _671-693_as 
                 hcmr 
               
               
                   
                   
                 UUCAAGGA 
                   
                   
                   
                 UUGGUCC 
                   
                   
                   
               
               
                   
               
               
                 AD-69384 
                 A-139705 
                 GGGCUUCAAGG 
                 65 
                 _678-698_s 
                 A-139706 
                 UCUCCUGAGGCC 
                 154 
                 _676-698_as 
                 hc 
               
               
                   
                   
                 CCUCACGAGA 
                   
                   
                   
                 UUGAACCCCUU 
                   
                   
                   
               
               
                   
               
               
                 AD-69429 
                 A-139794 
                 GCUUCAAGGCC 
                 66 
                 _682-702_s 
                 A-139795 
                 UCUCCUGAGGCC 
                 155 
                 _680-702_as 
                 hc 
               
               
                   
                   
                 UCAGGAGA 
                   
                   
                   
                 UUGAAGC 
                   
                   
                   
               
               
                   
               
               
                 AD-69385 
                 A-139707 
                 CAGGAGCAGAA 
                 67 
                 _692-712_s 
                 A-139708 
                 UAUUGUUCCCUU 
                 156 
                 _690-712_as 
                 hc 
               
               
                   
                   
                 GGGAACAAUA 
                   
                   
                   
                 CUGCUCCUGAG 
                   
                   
                   
               
               
                   
               
               
                 AD-69386 
                 A-139709 
                 GGAGCAGAAGG 
                 68 
                 _694-714_s 
                 A-139710 
                 UACAUUGUUCCC 
                 157 
                 _692-714_as 
                 hc 
               
               
                   
                   
                 GAACAAUGUA 
                   
                   
                   
                 UUCUGCUCCUG 
                   
                   
                   
               
               
                   
               
               
                 AD-69430 
                 A-1397% 
                 GGAGCAGAAGG 
                 69 
                 _696-716_s 
                 A-139797 
                 UAUUGUUCCCUU 
                 158 
                 _694-716_as 
                 hc 
               
               
                   
                   
                 GAACAAUA 
                   
                   
                   
                 CUGCUCC 
                   
                   
                   
               
               
                   
               
               
                 AD-69387 
                 A-1397II 
                 GAGCAGAAGGG 
                 70 
                 _695-715_s 
                 A-139712 
                 UGACAUUGUUCC 
                 159 
                 _693-715_as 
                 hc 
               
               
                   
                   
                 AACAAUGUCA 
                   
                   
                   
                 CUUCUGCUCCU 
                   
                   
                   
               
               
                   
               
               
                 AD-69431 
                 A-139798 
                 AGCAGAAGGGA 
                 71 
                 _698-718_s 
                 A-139799 
                 UACAUUGUUCCC 
                 160 
                 _696-718_as 
                 hc 
               
               
                   
                   
                 ACAAUGUA 
                   
                   
                   
                 UUCUGCU 
                   
                   
                   
               
               
                   
               
               
                 AD-69432 
                 A-139800 
                 GCAGAAGGGAA 
                 72 
                 _699-719_s 
                 A-139801 
                 UGACAUUGUUCC 
                 161 
                 _697-719_as 
                 hc 
               
               
                   
                   
                 CAAUGUCA 
                   
                   
                   
                 CUUCUGC 
                   
                   
                   
               
               
                   
               
               
                 AD-69388 
                 A-139713 
                 GACUUUGAAAU 
                 73 
                 _751-771_s 
                 A-139714 
                 UACCACAUCCAU 
                 162 
                 _749-771_as 
                 hcmr 
               
               
                   
                   
                 GGAUGUGGUA 
                   
                   
                   
                 UUCAAAGUCCC 
                   
                   
                   
               
               
                   
               
               
                 AD-69389 
                 A-139715 
                 CUUUGAAAUGG 
                 74 
                 _753-773_s 
                 A-139716 
                 UCCACCACAUCC 
                 163 
                 _751-773_as 
                 hcmr 
               
               
                   
                   
                 AUGUGGUGGA 
                   
                   
                   
                 AUUUCAAAGUC 
                   
                   
                   
               
               
                   
               
               
                 AD-69433 
                 A-139802 
                 CUUUGAAAUGG 
                 75 
                 _755-775_s 
                 A-139803 
                 UACCACAUCCAU 
                 164 
                 _753-775_as 
                 hcmr 
               
               
                   
                   
                 AUGUGGUA 
                   
                   
                   
                 UUCAAAG 
                   
                   
                   
               
               
                   
               
               
                 AD-69434 
                 A-139804 
                 UUGAAAUGGAU 
                 76 
                 _757-777_s 
                 A-139805 
                 UCCACCACAUCC 
                 165 
                 _755-777_as 
                 hcmr 
               
               
                   
                   
                 GUGGUGGA 
                   
                   
                   
                 AUUUCAA 
                   
                   
                   
               
               
                   
               
               
                 AD-69391 
                 A-139719 
                 UGAAAUGGAUG 
                 77 
                 _756-776_s 
                 A-139720 
                 AUUGCCACCACA 
                 166 
                 _754-776_as 
                 hcmr 
               
               
                   
                   
                 UGGUGGCAAU 
                   
                   
                   
                 UCCAUUUCAAA 
                   
                   
                   
               
               
                   
               
               
                 AD-69436 
                 A-139808 
                 AAAUGGAUGUG 
                 78 
                 _760-780_s 
                 A-139809 
                 AUUGCCACCACA 
                 167 
                 _758-780_as 
                 hcmr 
               
               
                   
                   
                 GUGGCAAU 
                   
                   
                   
                 UCCAUUU 
                   
                   
                   
               
               
                   
               
               
                 AD-69392 
                 A-139721 
                 AUGGAUGUGGU 
                 79 
                 _760-780_s 
                 A-139722 
                 UACCAUUGCCAC 
                 168 
                 _758-780_as 
                 hcmr 
               
               
                   
                   
                 GGCAAUGGUA 
                   
                   
                   
                 CACAUCCAUUU 
                   
                   
                   
               
               
                   
               
               
                 AD-69402 
                 A-139721 
                 AUGGAUGUGGU 
                 80 
                 _760-780_s 
                 A-139741 
                 UACCAUUGCCAC 
                 169 
                 _758-780_as 
                 mr 
               
               
                   
                   
                 GGCAAUGGUA 
                   
                   
                   
                 CACAUCCAUCU 
                   
                   
                   
               
               
                   
               
               
                 AD-69393 
                 A-139723 
                 GGAUGUGGUGG 
                 81 
                 _762-782_s 
                 A-139724 
                 UUCACCAUUGCC 
                 170 
                 _760-782_as 
                 hcmr 
               
               
                   
                   
                 CAAUGGUGAA 
                   
                   
                   
                 ACCACAUCCAU 
                   
                   
                   
               
               
                   
               
               
                 AD-69437 
                 A-139810 
                 GGAUGUGGUGG 
                 82 
                 _764-784_s 
                 A-1398II 
                 UACCAUUGCCAC 
                 171 
                 _762-784_as 
                 hcmr 
               
               
                   
                   
                 CAAUGGUA 
                   
                   
                   
                 CACAUCC 
                   
                   
                   
               
               
                   
               
               
                 AD-69438 
                 A-139812 
                 AUGUGGUGGCA 
                 83 
                 _766-786_s 
                 A-139813 
                 UUCACCAUUGCC 
                 172 
                 _764-786_as 
                 hcmr 
               
               
                   
                   
                 AUGGUGAA 
                   
                   
                   
                 ACCACAU 
                   
                   
                   
               
               
                   
               
               
                 AD-69395 
                 A-139727 
                 GCAAUGGUGAA 
                 84 
                 _772-792_s 
                 A-139728 
                 UACCGUGUCAUU 
                 173 
                 _770-792_as 
                 hc 
               
               
                   
                   
                 UGACACGGUA 
                   
                   
                   
                 CACCAUUGCCA 
                   
                   
                   
               
               
                   
               
               
                 AD-69440 
                 A-139816 
                 AAUGGUGAAUG 
                 85 
                 _776-796_s 
                 A-139817 
                 UACCGUGUCAUU 
                 174 
                 _774-796_as 
                 hc 
               
               
                   
                   
                 ACACGGUA 
                   
                   
                   
                 CACCAUU 
                   
                   
                   
               
               
                   
               
               
                 AD-69396 
                 A-139729 
                 CAGUGCGAGGU 
                 86 
                 _826-846_s 
                 A-139730 
                 UAUCAUGCCGAC 
                 175 
                 _824-846_as 
                 hcmr 
               
               
                   
                   
                 CGGCAUGAUA 
                   
                   
                   
                 CUCGCACUGAU 
                   
                   
                   
               
               
                   
               
               
                 AD-69441 
                 A-139818 
                 GUGCGAGGUCG 
                 87 
                 _830-850_s 
                 A-139819 
                 UAUCAUGCCGAC 
                 176 
                 _828-850_as 
                 hcmr 
               
               
                   
                   
                 GCAUGAUA 
                   
                   
                   
                 CUCGCAC 
                   
                   
                   
               
               
                   
               
               
                 AD-69397 
                 A-139731 
                 ACAUGGAGGAG 
                 88 
                 _872-892_s 
                 A-139732 
                 UAUUCUGCAUCU 
                 177 
                 _870-892_as 
                 hc 
               
               
                   
                   
                 AUGCAGAAUA 
                   
                   
                   
                 CCUCCAUGUAG 
                   
                   
                   
               
               
                   
               
               
                 AD-69442 
                 A-139820 
                 AUGGAGGAGAU 
                 89 
                 _876-896_s 
                 A-139821 
                 UAUUCUGCAUCU 
                 178 
                 _874-896_as 
                 hc 
               
               
                   
                   
                 GCAGAAUA 
                   
                   
                   
                 CCUCCAU 
                   
                   
                   
               
               
                   
               
               
                 AD-69394 
                 A-139725 
                 GAUGCAGAAUG 
                 90 
                 _882-902_s 
                 A-139726 
                 ACCAGCUCCACA 
                 179 
                 _880-902_as 
                 hcmr 
               
               
                   
                   
                 UGGAGCUGGU 
                   
                   
                   
                 UUCUGCAUCUC 
                   
                   
                   
               
               
                   
               
               
                 AD-69439 
                 A-139814 
                 UGCAGAAUGUG 
                 91 
                 _886-906_s 
                 A-139815 
                 ACCAGCUCCACA 
                 180 
                 _884-906_as 
                 hcmr 
               
               
                   
                   
                 GAGCUGGU 
                   
                   
                   
                 UUCUGCA 
                   
                   
                   
               
               
                   
               
               
                 AD-69398 
                 A-139733 
                 GUGGACGAGAG 
                 92 
                 _1000-1020_s 
                 A-139734 
                 UUUUGCAGAGCU 
                 181 
                  _998-1020_as 
                 hc 
               
               
                   
                   
                 CUCUGCAAAA 
                   
                   
                   
                 CUCGUCCACCA 
                   
                   
                   
               
               
                   
               
               
                 AD-69443 
                 A-139822 
                 GGACGAGAGCU 
                 93 
                 _1004-1024_s 
                 A-139823 
                 UUUUGCAGAGCU 
                 182 
                 _1002-1024_as 
                 hc 
               
               
                   
                   
                 CUGCAAAA 
                   
                   
                   
                 CUCGUCC 
                   
                   
                   
               
               
                   
               
               
                 AD-69380 
                 A-139697 
                 AAGUACAUGGG 
                 94 
                 _1057-1077_s 
                 A-139698 
                 UACCAGCUCGCC 
                 183 
                 _1055-1077_as 
                 hcmr 
               
               
                   
                   
                 CGAGCUGGUA 
                   
                   
                   
                 CAUGUACUUGC 
                   
                   
                   
               
               
                   
               
               
                 AD-69425 
                 A-139786 
                 GUACAUGGGCG 
                 95 
                 _1061-1081_s 
                 A-139787 
                 UACCAGCUCGCC 
                 184 
                 _1059-1081_as 
                 hcmr 
               
               
                   
                   
                 AGCUGGUA 
                   
                   
                   
                 CAUGUAC 
                   
                   
                   
               
               
                   
               
               
                 AD-69403 
                 A-139742 
                 AGGCUCGUGGA 
                 96 
                 _1093-1113_s 
                 A-139743 
                 UAGGUUUUCGUC 
                 185 
                 _1091-1113_as 
                 hc 
               
               
                   
                   
                 CGAAAACCUA 
                   
                   
                   
                 CACGAGCCUGA 
                   
                   
                   
               
               
                   
               
               
                 AD-69447 
                 A-139830 
                 GCUCGUGGACG 
                 97 
                 _1097-1117_s 
                 A-139831 
                 UAGGUUUUCGUC 
                 186 
                 _1095-1117_as 
                 hc 
               
               
                   
                   
                 AAAACCUA 
                   
                   
                   
                 CACGAGC 
                   
                   
                   
               
               
                   
               
               
                 AD-69404 
                 A-139744 
                 CGUGGACGAAA 
                 98 
                 _1098-1118_s 
                 A-139745 
                 AAGAGCAGGUUU 
                 187 
                 _1096-1118_as 
                 hc 
               
               
                   
                   
                 ACCUGCUCUU 
                   
                   
                   
                 UCGUCCACGAG 
                   
                   
                   
               
               
                   
               
               
                 AD-69405 
                 A-139746 
                 GUGGACGAAAA 
                 99 
                 _1099-1119_s 
                 A-139747 
                 UAAGAGCAGGUU 
                 188 
                 _1097-1119_as 
                 hc 
               
               
                   
                   
                 CCUGCUCUUA 
                   
                   
                   
                 UUCGUCCACGA 
                   
                   
                   
               
               
                   
               
               
                 AD-69448 
                 A-139832 
                 UGGACGAAAAC 
                 100 
                 _1102-1122_s 
                 A-139833 
                 AAGAGCAGGUUU 
                 189 
                 _1100-1122_as 
                 hc 
               
               
                   
                   
                 CUGCUCUU 
                   
                   
                   
                 UCGUCCA 
                   
                   
                   
               
               
                   
               
               
                 AD-69449 
                 A-139834 
                 GGACCAAAACC 
                 101 
                 _1103-1123_s 
                 A-139835 
                 UAAGAGCAGGUU 
                 190 
                 _1101-1123_as 
                 hc 
               
               
                   
                   
                 UGCUCUUA 
                   
                   
                   
                 UUCGUCC 
                   
                   
                   
               
               
                   
               
               
                 AD-69406 
                 A-139748 
                 CGCAAGCAGAU 
                 102 
                 _1204-1224_s 
                 A-139749 
                 UAUGUUGUAGAU 
                 191 
                 _1202-1224_as 
                 hc 
               
               
                   
                   
                 CUACAACAUA 
                   
                   
                   
                 CUGCUUGCGGU 
                   
                   
                   
               
               
                   
               
               
                 AD-69450 
                 A-139836 
                 CAAGCAGAUCU 
                 103 
                 _1208-1228_s 
                 A-139837 
                 UAUGUUGUAGAU 
                 192 
                 _1206-1228_as 
                 hc 
               
               
                   
                   
                 ACAACAUA 
                   
                   
                   
                 CUGCUUG 
                   
                   
                   
               
               
                   
               
               
                 AD-69390 
                 A-139717 
                 AGCUGCGAGAU 
                 104 
                 _1468-1488_s 
                 A-139718 
                 UAUGAAGGUGAU 
                 193 
                 _1466-1488_as 
                 hcmr 
               
               
                   
                   
                 CACCUUCAUA 
                   
                   
                   
                 CUCGCAGCUGG 
                   
                   
                   
               
               
                   
               
               
                 AD-69435 
                 A-139806 
                 CUGCGAGAUCA 
                 105 
                 _1472-1492_s 
                 A-139807 
                 UAUGAAGGUGAU 
                 194 
                 _1470-1492_as 
                 hcmr 
               
               
                   
                   
                 CCUUCAUA 
                   
                   
                   
                 CUCGCAG 
                   
                   
                   
               
               
                   
               
               
                 AD-69408 
                 A-139752 
                 CCAGUCCUGGC 
                 106 
                 _2049-2069_s 
                 A-139753 
                 UAAGAAAAUGGC 
                 195 
                 _2047-2069_as 
                 hc 
               
               
                   
                   
                 CAUUUUCUUA 
                   
                   
                   
                 CAGGACUGGGU 
                   
                   
                   
               
               
                   
               
               
                 AD-69452 
                 A-139840 
                 AGUCCUGGCCA 
                 107 
                 _2053-2073_s 
                 A-139841 
                 UAAGAAAAUGGC 
                 196 
                 _2051-2073_as 
                 hc 
               
               
                   
                   
                 UUUUCUUA 
                   
                   
                   
                 CAGGACU 
                   
                   
                   
               
               
                   
               
               
                 AD-69409 
                 A-139754 
                 CACUGAGUGGC 
                 108 
                 _2170-2190_s 
                 A-139755 
                 AGAAUCACAAGC 
                 197 
                 _2168-2190_as 
                 hc 
               
               
                   
                   
                 UUGUGAUUCU 
                   
                   
                   
                 CACUCAGUGAU 
                   
                   
                   
               
               
                   
               
               
                 AD-69453 
                 A-139842 
                 CUGAGUGGCUU 
                 109 
                 _2174-2194_s 
                 A-139843 
                 AGAAUCACAAGC 
                 198 
                 _2172-2194_as 
                 hc 
               
               
                   
                   
                 GUGAUUCU 
                   
                   
                   
                 CACUCAG 
                   
                   
                   
               
               
                   
               
               
                 AD-69410 
                 A-139756 
                 AAUGUUAAAAG 
                 110 
                 _2413-2433_s 
                 A-139757 
                 AUGUUUAAAACU 
                 199 
                 _2411-2433_as 
                 hc 
               
               
                   
                   
                 UUUUAAACAU 
                   
                   
                   
                 UUUAACAUUUU 
                   
                   
                   
               
               
                   
               
               
                 AD-69454 
                 A-139844 
                 UGUUAAAAGUU 
                 111 
                 _2417-2437_s 
                 A-139845 
                 AUGUUUAAAACU 
                 200 
                 _2415-2437_as 
                 hc 
               
               
                   
                   
                 UUAAACAU 
                   
                   
                   
                 UUUAACA 
                   
                   
                   
               
               
                   
               
               
                 AD-69444 
                 A-139824 
                 AGCAGAAGGGA 
                 112 
                 _698-718_s 
                 A-139825 
                 UAUGUUGUUCCC 
                 201 
                 _696-718_as 
                 mr 
               
               
                   
                   
                 ACAACAUA 
                   
                   
                   
                 UUCUGCU 
                   
                   
                   
               
               
                   
               
               
                 AD-69399 
                 A-139735 
                 GGAGCAGAAGG 
                 113 
                 _694-714_s 
                 A-139736 
                 UAUGUUGUUCCC 
                 202 
                 _692-714_as 
                 mr 
               
               
                   
                   
                 GAACAACAUA 
                   
                   
                   
                 UUCUGCUCCGG 
                   
                   
                   
               
               
                   
               
               
                 AD-69445 
                 A-139826 
                 UCUCCGAGAUG 
                 114 
                 _725-745_s 
                 A-139827 
                 UUUGAUAGCAUC 
                 203 
                 _723-745_as 
                 mr 
               
               
                   
                   
                 CUAUCAAA 
                   
                   
                   
                 UCGGAGA 
                   
                   
                   
               
               
                   
               
               
                 AD-69400 
                 A-139737 
                 CUUCUCCGAGA 
                 115 
                 _721-741_s 
                 A-139738 
                 UUUGAUAGCAUC 
                 204 
                 _719-741_as 
                 mr 
               
               
                   
                   
                 UGCUAUCAAA 
                   
                   
                   
                 UCGGAGAAGUC 
                   
                   
                   
               
               
                   
               
               
                 AD-69446 
                 A-139828 
                 AGAUGGAUGUG 
                 116 
                 _760-780_s 
                 A-139829 
                 AUUGCCACCACA 
                 205 
                 _758-780_as 
                 mr 
               
               
                   
                   
                 GUGGCAAU 
                   
                   
                   
                 UCCAUCU 
                   
                   
                   
               
               
                   
               
               
                 AD-69401 
                 A-139739 
                 UGAGAUGGAUG 
                 117 
                 _756-776_s 
                 A-139740 
                 AUUGCCACCACA 
                 206 
                 _754-776_as 
                 mr 
               
               
                   
                   
                 UGGUGGCAAU 
                   
                   
                   
                 UCCAUCUCAAA 
                   
                   
                   
               
               
                   
               
               
                 AD-69451 
                 A-139838 
                 CUGCGAAAUCA 
                 118 
                 _1472-1492_s 
                 A-139839 
                 AAUGAAGGUGAU 
                 207 
                 _1470-1492_as 
                 mr 
               
               
                   
                   
                 CCUUCAUU 
                   
                   
                   
                 UUCGCAG 
                   
                   
                   
               
               
                   
               
               
                 AD-69407 
                 A-139750 
                 AACUGCGAAAU 
                 119 
                 _1468-1488_s 
                 A-139751 
                 AAUGAAGGUGAU 
                 208 
                 _1466-1488_as 
                 mr 
               
               
                   
                   
                 CACCUUCAUU 
                   
                   
                   
                 UUCGCAGUUGG 
               
               
                   
               
            
           
         
       
     
     
       
         
           
               
             
               
                 TABLE 3 
               
             
            
               
                   
               
               
                 GCK Modified Sequences 
               
            
           
           
               
               
               
               
               
               
               
               
               
            
               
                   
                   
                 Sense 
                 SEQ 
                 Anti- 
                 Antisense 
                 SEQ 
                   
                 SEQ 
               
               
                 Duplex 
                 Sense 
                 sequence 
                 ID 
                 sense 
                 sequence 
                 ID 
                 mRNA 
                 ID 
               
               
                 ID 
                 ID 
                 (5′-3′) 
                 NO: 
                 ID 
                 (5′-3′) 
                 NO: 
                 sequence 
                 NO: 
               
               
                   
               
               
                 AD-69366 
                 A-139669 
                 gsasggacCfuGfAf 
                 209 
                 A-139670 
                 usAfsucaCfcUfUfcu 
                 298 
                 AGGAGGACCUGA 
                 387 
               
               
                   
                   
                 AfgaaggugauaL96 
                   
                   
                 ucAfgGfuccucscsu 
                   
                 AGAAGGUGAUA 
                   
               
               
                   
               
               
                 AD-69368 
                 A-139673 
                 gsgsaccuGfaAfGf 
                 210 
                 A-139674 
                 usUfscauCfaCfCfuu 
                 299 
                 GAGGACCUGAAG 
                 388 
               
               
                   
                   
                 AfaggugaugaaL96 
                   
                   
                 cuUfcAfgguccsusc 
                   
                 AAGGUGAUGAA 
                   
               
               
                   
               
               
                 AD-69411 
                 A-139758 
                 GGACCUGAAGAAGGU 
                 211 
                 A-139759 
                 UAUCACCUUCUUCAGGU 
                 300 
                 GGACCUGAAGAA 
                 389 
               
               
                   
                   
                 GAUAdTdT 
                   
                   
                 CCdTdT 
                   
                 GGUGAUA 
                   
               
               
                   
               
               
                 AD-69413 
                 A-139762 
                 ACCUGAAGAAGGUGA 
                 212 
                 A-139763 
                 UUCAUCACCUUCUUCAG 
                 301 
                 ACCUGAAGAAGG 
                 390 
               
               
                   
                   
                 UGAAdTdT 
                   
                   
                 GUdTdT 
                   
                 UGAUGAA 
                   
               
               
                   
               
               
                 AD-69367 
                 A-139671 
                 gsgsugauGfaGfAf 
                 213 
                 A-139672 
                 usUfscugCfaUfCfcg 
                 302 
                 AAGGUGAUGAGA 
                 391 
               
               
                   
                   
                 CfggaugcagaaL96 
                   
                   
                 ucUfcAfucaccsusu 
                   
                 CGGAUGCAGAA 
                   
               
               
                   
               
               
                 AD-69369 
                 A-139675 
                 usgsaugaGfaCfGf 
                 214 
                 A-139676 
                 usCfsuucUfgCfAfuc 
                 303 
                 GGUGAUGAGACG 
                 392 
               
               
                   
                   
                 GfaugcagaagaL96 
                   
                   
                 cgUfcUfcaucascsc 
                   
                 GAUGCAGAAGA 
                   
               
               
                   
               
               
                 AD-69412 
                 A-139760 
                 UGAUGAGACGGAUGC 
                 215 
                 A-139761 
                 UUCUGCAUCCGUCUCAU 
                 304 
                 UGAUGAGACGGA 
                 393 
               
               
                   
                   
                 AGAAdTdT 
                   
                   
                 CAdTdT 
                   
                 UGCAGAA 
                   
               
               
                   
               
               
                 AD-69414 
                 A-139764 
                 AUGAGACGGAUGCAG 
                 216 
                 A-139765 
                 UCUUCUGCAUCCGUCUC 
                 305 
                 AUGAGACGGAUG 
                 394 
               
               
                   
                   
                 AAGAdTdT 
                   
                   
                 AUdTdT 
                   
                 CAGAAGA 
                   
               
               
                   
               
               
                 AD-69371 
                 A-139679 
                 ascsggauGfcAfGf 
                 217 
                 A-139680 
                 usCfscauCfuCrCfuu 
                 306 
                 AGACGGAUGCAG 
                 395 
               
               
                   
                   
                 AfaggagauggaL96 
                   
                   
                 cuGfcAfuccguscsu 
                   
                 AAGGAGAUGGA 
                   
               
               
                   
               
               
                 AD-69416 
                 A-139768 
                 GGAUGCAGAAGGAGA 
                 218 
                 A-139769 
                 UCCAUCUCCUUCUGCAU 
                 307 
                 GGAUGCAGAAGG 
                 396 
               
               
                   
                   
                 UGGAdTdT 
                   
                   
                 CCdTdT 
                   
                 AGAUGGA 
                   
               
               
                   
               
               
                 AD-69370 
                 A-139677 
                 ascsccauGfaAfGf 
                 219 
                 A-139678 
                 usAfscacUfgGfCfcu 
                 308 
                 AGACCCAUGAAG 
                 397 
               
               
                   
                   
                 ATggccaguguaL96 
                   
                   
                 cuUfcAfuggguscsu 
                   
                 AGGCCAGUGUA 
                   
               
               
                   
               
               
                 AD-69415 
                 A-139766 
                 CCAUGAAGAGGCCAG 
                 220 
                 A-139767 
                 UACACUGGCCUCUUCAU 
                 309 
                 CCAUGAAGAGGC 
                 398 
               
               
                   
                   
                 UGUAdTdT 
                   
                   
                 GGdTdT 
                   
                 CAGUGUA 
                   
               
               
                   
               
               
                 AD-69372 
                 A-139681 
                 csasguguGfaAfGf 
                 221 
                 A-139682 
                 usUfsgggCfaGfCfau 
                 310 
                 GCCAGUGUGAAG 
                 399 
               
               
                   
                   
                 AfugcugcccaaL96 
                   
                   
                 cuUfcAfcacugsgsc 
                   
                 AUGCUGCCCAA 
                   
               
               
                   
               
               
                 AD-69417 
                 A-139770 
                 GUGUGAAGAUGCUGC 
                 222 
                 A-139771 
                 UUGGGCAGCAUCUUCAC 
                 311 
                 GUGUGAAGAUGC 
                 400 
               
               
                   
                   
                 CCAAdTdT 
                   
                   
                 ACdTdT 
                   
                 UGCCCAA 
                   
               
               
                   
               
               
                 AD-69373 
                 A-139683 
                 gsusgaagGfuGfGf 
                 223 
                 A-139684 
                 usUfscacCfuUfCfuc 
                 312 
                 UGGUGAAGGUGG 
                 401 
               
               
                   
                   
                 GfagaaggugaaL96 
                   
                   
                 ccAfcCfuucacscsa 
                   
                 GAGAAGGUGAA 
                   
               
               
                   
               
               
                 AD-69418 
                 A-139772 
                 GAAGGUGGGAGAAGG 
                 224 
                 A-139773 
                 UUCACCUUCUCCCACCU 
                 313 
                 GAAGGUGGGAGA 
                 402 
               
               
                   
                   
                 UGAAdTdT 
                   
                   
                 UCdTdT 
                   
                 AGGUGAA 
                   
               
               
                   
               
               
                 AD-69374 
                 A-139685 
                 gsasgcguGfaAfGf 
                 225 
                 A-139686 
                 usGfsgugUfuUfGfgu 
                 314 
                 UGGAGCGUGAAG 
                 403 
               
               
                   
                   
                 AfccaaacaccaL96 
                   
                   
                 cuUfcAfcgcucscsa 
                   
                 ACCAAACACCA 
                   
               
               
                   
               
               
                 AD-69419 
                 A-139774 
                 GCGUGAAGACCAAAC 
                 226 
                 A-139775 
                 UGGUGUUUGGLCUUCAC 
                 315 
                 GCGUGAAGACCA 
                 404 
               
               
                   
                   
                 ACCAdTdT 
                   
                   
                 GCdTdT 
                   
                 AACACCA 
                   
               
               
                   
               
               
                 AD-69375 
                 A-139687 
                 gsasagacCfaAfAf 
                 227 
                 A-139688 
                 usAfscauCfuGfGfug 
                 316 
                 GUGAAGACCAAA 
                 405 
               
               
                   
                   
                 CfaccagauguaL96 
                   
                   
                 uuUfgGfucuucsasc 
                   
                 CACCAGAUGUA 
                   
               
               
                   
               
               
                 AD-69420 
                 A-139776 
                 AGACCAAACACCAGA 
                 228 
                 A-139777 
                 UACAUCUGGUGUUUGGU 
                 317 
                 AGACCAAACACC 
                 406 
               
               
                   
                   
                 UGUAdTdT 
                   
                   
                 CUdTdT 
                   
                 AGAUGUA 
                   
               
               
                   
               
               
                 AD-69376 
                 A-139689 
                 gsascuucCTuGfGf 
                 229 
                 A-139690 
                 usUfsgauGfcUfUfgu 
                 318 
                 CCGACUUCCUGG 
                 407 
               
               
                   
                   
                 AfcaagcaucaaL96 
                   
                   
                 ccAfgGfaagucsgsg 
                   
                 ACAAGCAUCAA 
                   
               
               
                   
               
               
                 AD-69377 
                 A-139691 
                 csusuccuGfgAfCf 
                 230 
                 A-139692 
                 asUfscugAfuGfCfuu 
                 319 
                 GACUUCCUGGAC 
                 408 
               
               
                   
                   
                 AfagcaucagauL96 
                   
                   
                 guCfcAfggaagsusc 
                   
                 AAGCAUCAGAU 
                   
               
               
                   
               
               
                 AD-69421 
                 A-139778 
                 CUUCCUGGACAAGCA 
                 231 
                 A-139779 
                 UCGAUGCUUGUCCAGGA 
                 320 
                 CUUCCUGGACAA 
                 409 
               
               
                   
                   
                 UCAAdTdT 
                   
                   
                 AGdTdT 
                   
                 GCAUCAA 
                   
               
               
                   
               
               
                 AD-69422 
                 A-139780 
                 UCCUGGACAAGCAUC 
                 232 
                 A-139781 
                 AUCUGAUGCCUGUCCAG 
                 321 
                 UCCUGGACAAGC 
                 410 
               
               
                   
                   
                 AGAUdTdT 
                   
                   
                 GAdTdT 
                   
                 AUCAGAU 
                   
               
               
                   
               
               
                 AD-69378 
                 A-139693 
                 cscsuggaCfaAfGf 
                 233 
                 A-139694 
                 usUfscauCfuGfAfug 
                 322 
                 UUCCUGGACAAG 
                 411 
               
               
                   
                   
                 CfaucagaugaaL96 
                   
                   
                 cuUfgUfccaggsasa 
                   
                 CAUCAGAUGAA 
                   
               
               
                   
               
               
                 AD-69379 
                 A-139695 
                 csusggacAfaGfCf 
                 234 
                 A-13%% 
                 usUfsucaUfcUfGfau 
                 323 
                 UCCUGGACAAGC 
                 412 
               
               
                   
                   
                 AfucagaugaaaL96 
                   
                   
                 gcUfuGfuccagsgsa 
                   
                 AUCAGAUGAAA 
                   
               
               
                   
               
               
                 AD-69423 
                 A-139782 
                 UGGACAAGCAUCAGA 
                 235 
                 A-139783 
                 UUCAUCUGAUGCUUGUC 
                 324 
                 UGGACAAGCAUC 
                 413 
               
               
                   
                   
                 UGAAdTdT 
                   
                   
                 CAdTdT 
                   
                 AGAUGAA 
                   
               
               
                   
               
               
                 AD-69424 
                 A-139784 
                 GGACAAGCAUCAGAC 
                 236 
                 A-139785 
                 UUUCAUCUGAUGCUUGU 
                 325 
                 GGACAAGCAUCA 
                 414 
               
               
                   
                   
                 GAAAdTdT 
                   
                   
                 CCdTdT 
                   
                 GAUGAAA 
                   
               
               
                   
               
               
                 AD-69381 
                 A-139699 
                 asgsgcacGfaAfGf 
                 237 
                 A-139700 
                 usUfsuauCfgAfUfgu 
                 326 
                 UGAGGCACGAAG 
                 415 
               
               
                   
                   
                 AfcaucgauaaaL96 
                   
                   
                 cuUfcGfugccuscsa 
                   
                 ACAUCGAUAAA 
                   
               
               
                   
               
               
                 AD-69426 
                 A-139788 
                 GCACGAAGACAUCGA 
                 238 
                 A-139789 
                 UUUAUCGAUGUCUUCGU 
                 327 
                 GCACGAAGACAU 
                 416 
               
               
                   
                   
                 UAAAdTdT 
                   
                   
                 GCdTdT 
                   
                 CGAUAAA 
                   
               
               
                   
               
               
                 AD-69382 
                 A-139701 
                 ascsaucgAfuAfAf 
                 239 
                 A-139702 
                 usAfsaggAfuGfCfcc 
                 328 
                 AGACAUCGAUAA 
                 417 
               
               
                   
                   
                 GfggcauccuuaL96 
                   
                   
                 uuAfuCfgauguscsu 
                   
                 GGGCAUCCUUA 
                   
               
               
                   
               
               
                 AD-69427 
                 A-139790 
                 AUCGAUAAGGGCAUC 
                 240 
                 A-139791 
                 UAAGGAUGCCCUUAUCG 
                 329 
                 AUCGAUAAGGGC 
                 418 
               
               
                   
                   
                 CUUAdTdT 
                   
                   
                 AUdTdT 
                   
                 AUCCUUA 
                   
               
               
                   
               
               
                 AD-69383 
                 A-139703 
                 csusggacCfaAfGf 
                 241 
                 A-139704 
                 usCfscuuGfaAfGfcc 
                 330 
                 AACUGGACCAAG 
                 419 
               
               
                   
                   
                 GfgcuucaaggaL96 
                   
                   
                 cuUfgGfuccagsusu 
                   
                 GGCUUCAAGGA 
                   
               
               
                   
               
               
                 AD-69428 
                 A-139792 
                 GGACCAAGGGCLUCA 
                 242 
                 A-139793 
                 UCCUUGAAGCCCUUGGU 
                 331 
                 GGACCAAGGGCU 
                 420 
               
               
                   
                   
                 AGGAdTdT 
                   
                   
                 CCdTdT 
                   
                 UCAAGGA 
                   
               
               
                   
               
               
                 AD-69384 
                 A-139705 
                 gsgsgcuuCfaAfGf 
                 243 
                 A-139706 
                 usCfsuccUfgAfGfgc 
                 332 
                 AAGGGCUUCAAG 
                 421 
               
               
                   
                   
                 GfccucaggagaL96 
                   
                   
                 cuUfgAfagcccsusu 
                   
                 GCCUCAGGAGA 
                   
               
               
                   
               
               
                 AD-69429 
                 A-139794 
                 GCUUCAAGGCCUCAG 
                 244 
                 A-139795 
                 UCUCCUGAGGCCUUGAA 
                 333 
                 GCUUCAAGGCCU 
                 422 
               
               
                   
                   
                 GAGAdTdT 
                   
                   
                 GCdTdT 
                   
                 CAGGAGA 
                   
               
               
                   
               
               
                 AD-69385 
                 A-139707 
                 csasggagCfaGfAf 
                 245 
                 A-139708 
                 usAfsuugUfuCfCfcu 
                 334 
                 CUCAGGAGCAGA 
                 423 
               
               
                   
                   
                 AfgggaacaauaL96 
                   
                   
                 ucUfgCfuccugsasg 
                   
                 AGGGAACAAUA 
                   
               
               
                   
               
               
                 AD-69386 
                 A-139709 
                 gsgsagcaGfaAfGf 
                 246 
                 A-139710 
                 usAfscauUfgUfUfcc 
                 335 
                 CAGGAGCAGAAG 
                 424 
               
               
                   
                   
                 GfgaacaauguaL96 
                   
                   
                 cuUfcUfgcuccsusg 
                   
                 GGAACAAUGUA 
                   
               
               
                   
               
               
                 AD-69430 
                 A-139796 
                 GGAGCAGAAGGGAAC 
                 247 
                 A-139797 
                 UAUUGUUCCCUUCUGCU 
                 336 
                 GGAGCAGAAGGG 
                 425 
               
               
                   
                   
                 AAUAdTdT 
                   
                   
                 CCdTdT 
                   
                 AACAAUA 
                   
               
               
                   
               
               
                 AD-69387 
                 A-139711 
                 gsasgcagAfaGfGf 
                 248 
                 A-139712 
                 usGfsacaUfuGfUfuc 
                 337 
                 AGGAGCAGAAGG 
                 426 
               
               
                   
                   
                 GfaacaaugucaL96 
                   
                   
                 ccUfuCfugcucscsu 
                   
                 GAACAAUGUCA 
                   
               
               
                   
               
               
                 AD-69431 
                 A-139798 
                 AGCAGAAGGGAACAA 
                 249 
                 A-139799 
                 UACAUUGUUCCCUUCUG 
                 338 
                 AGCAGAAGGGAA 
                 427 
               
               
                   
                   
                 UGUAdTdT 
                   
                   
                 CUdTdT 
                   
                 CAAUGUA 
                   
               
               
                   
               
               
                 AD-69432 
                 A-139800 
                 GCAGAAGGGAACAAU 
                 250 
                 A-139801 
                 UGACAUUGUUCCCUUCU 
                 339 
                 GCAGAAGGGAAC 
                 428 
               
               
                   
                   
                 GUCAdTdT 
                   
                   
                 GCdTdT 
                   
                 AAUGUCA 
                   
               
               
                   
               
               
                 AD-69388 
                 A-139713 
                 gsascuuuGfaAfAf 
                 251 
                 A-139714 
                 usAfsccaCfaUfCfca 
                 340 
                 GGGACUUUGAAA 
                 429 
               
               
                   
                   
                 UfggaugugguaL96 
                   
                   
                 uuUfcAfaagucscsc 
                   
                 UGGAUGUGGUA 
                   
               
               
                   
               
               
                 AD-69389 
                 A-139715 
                 csusuugaAfaUfGf 
                 252 
                 A-139716 
                 usCfscacCfaCfAfuc 
                 341 
                 GACUUUGAAAUG 
                 430 
               
               
                   
                   
                 GfaugugguggaL96 
                   
                   
                 caUfuUfcaaagsusc 
                   
                 GAUGUGGUGGA 
                   
               
               
                   
               
               
                 AD-69433 
                 A-139802 
                 CUUUGAAAUGGAUGU 
                 253 
                 A-139803 
                 UACCACAUCCAUUUCAA 
                 342 
                 CUUUGAAAUGGA 
                 431 
               
               
                   
                   
                 GGUAdTdT 
                   
                   
                 AGdTdT 
                   
                 UGUGGUA 
                   
               
               
                   
               
               
                 AD-69434 
                 A-139804 
                 UUGAAAUGGAUGUGG 
                 254 
                 A-139805 
                 UCCACCACAUCCAUUUC 
                 343 
                 UUGAAAUGGAUG 
                 432 
               
               
                   
                   
                 UGGAdTdT 
                   
                   
                 AAdTdT 
                   
                 UGGUGGA 
                   
               
               
                   
               
               
                 AD-69391 
                 A-139719 
                 usgsaaauGfgAfUf 
                 255 
                 A-139720 
                 asUfsugcCfaCfCfac 
                 344 
                 UUUGAAAUGGAU 
                 433 
               
               
                   
                   
                 GfugguggcaauL96 
                   
                   
                 auCfcAfuuucasasa 
                   
                 GUGGUGGCAAU 
                   
               
               
                   
               
               
                 AD-69436 
                 A-139808 
                 AAAUGGAUGUGGUGG 
                 256 
                 A-139809 
                 AUUGCCACCACAUCCAU 
                 345 
                 AAAUGGAUGUGG 
                 434 
               
               
                   
                   
                 CAAUdTdT 
                   
                   
                 UUdTdT 
                   
                 UGGCAAU 
                   
               
               
                   
               
               
                 AD-69392 
                 A-139721 
                 asusggauGfuGfCf 
                 257 
                 A-139722 
                 usAfsccaUfuGfCfca 
                 346 
                 AAAUGGAUGUGG 
                 435 
               
               
                   
                   
                 UfggcaaugguaL96 
                   
                   
                 ccAfcAfuccaususu 
                   
                 UGGCAAUGGUA 
                   
               
               
                   
               
               
                 AD-69402 
                 A-139721 
                 asusggauGfuGfGf 
                 258 
                 A-139741 
                 usAfsccaUfuGfCfca 
                 347 
                 AGAUGGAUGUGG 
                 436 
               
               
                   
                   
                 UfggcaaugguaL96 
                   
                   
                 ccAfcAfuccauscsu 
                   
                 UGGCAAUGGUA 
                   
               
               
                   
               
               
                 AD-69393 
                 A-139723 
                 gsgsauguGfgUfGf 
                 259 
                 A-139724 
                 usUfscacCfaUfUfgc 
                 348 
                 AUGGAUGUGGUG 
                 437 
               
               
                   
                   
                 CfcaauggugaaL96 
                   
                   
                 caCfcAfcauccsasu 
                   
                 GCAAUGGUGAA 
                   
               
               
                   
               
               
                 AD-69437 
                 A-139810 
                 GGAUGUGGUGGCAAU 
                 260 
                 A-139811 
                 UACCAUUGCCACCACAU 
                 349 
                 GGAUGUGGUGGC 
                 438 
               
               
                   
                   
                 GGUAdTdT 
                   
                   
                 CCdTdT 
                   
                 AAUGGUA 
                   
               
               
                   
               
               
                 AD-69438 
                 A-139812 
                 AUGUGGUGGCAAUGG 
                 261 
                 A-139813 
                 UUCACCAUUGCCACCAC 
                 350 
                 AUGUGGUGGCAA 
                 439 
               
               
                   
                   
                 UGAAdTdT 
                   
                   
                 AUdTdT 
                   
                 UGGUGAA 
                   
               
               
                   
               
               
                 AD-69395 
                 A-139727 
                 gscsaaugGfuGfAf 
                 262 
                 A-139728 
                 usAfsccgUfgUfCfau 
                 351 
                 UGGCAAUGGUGA 
                 440 
               
               
                   
                   
                 AfugacacgguaL96 
                   
                   
                 ucAfcCfauugcscsa 
                   
                 AUGACACGGUA 
                   
               
               
                   
               
               
                 AD-69440 
                 A-139816 
                 AAUGGUGAAUGACAC 
                 263 
                 A-139817 
                 UACCGUGLCAUUCACCA 
                 352 
                 AAUGGUGAAUGA 
                 441 
               
               
                   
                   
                 GGUAdTdT 
                   
                   
                 UUdTdT 
                   
                 CACGGUA 
                   
               
               
                   
               
               
                 AD-69396 
                 A-139729 
                 csasgugcGfaGfGf 
                 264 
                 A-139730 
                 usAfsucaUfgCfCfga 
                 353 
                 AUCAGUGCGAGG 
                 442 
               
               
                   
                   
                 UfcggcaugauaL96 
                   
                   
                 ccUfcGfcacugsasu 
                   
                 UCGGCAUGAUA 
                   
               
               
                   
               
               
                 AD-69441 
                 A-139818 
                 GUGCGAGGUCGGCAU 
                 265 
                 A-139819 
                 UAUCALGCCGACCUCGC 
                 354 
                 GUGCGAGGUCGG 
                 443 
               
               
                   
                   
                 GAUAdTdT 
                   
                   
                 ACdTdT 
                   
                 CAUGALA 
                   
               
               
                   
               
               
                 AD-69397 
                 A-139731 
                 ascsauggAfgGfAf 
                 266 
                 A-139732 
                 usAfsuucUfgCfAfuc 
                 355 
                 CUACAUGGAGGA 
                 444 
               
               
                   
                   
                 GfaugcagaauaL96 
                   
                   
                 ucCfuCfcaugusasg 
                   
                 GAUGCAGAAUA 
                   
               
               
                   
               
               
                 AD-69442 
                 A-139820 
                 AUGGAGGAGAUGCAG 
                 267 
                 A-139821 
                 UAUUCUGCAUCUCCUCC 
                 356 
                 AUGGAGGAGAUG 
                 445 
               
               
                   
                   
                 AAUAdTdT 
                   
                   
                 AUdTdT 
                   
                 CAGAAUA 
                   
               
               
                   
               
               
                 AD-69394 
                 A-139725 
                 gsasugcaGfaAfUf 
                 268 
                 A-139726 
                 asCfscagCfuCfCfac 
                 357 
                 GAGAUGCAGAAU 
                 446 
               
               
                   
                   
                 GfuggagcugguL96 
                   
                   
                 auUfcUfgcaucsusc 
                   
                 GUGGAGCUGGU 
                   
               
               
                   
               
               
                 AD-69439 
                 A-139814 
                 UGCAGAAUGUGGAGC 
                 269 
                 A-139815 
                 ACCAGCUCCACAUUCUG 
                 358 
                 UGCAGAAUGUGG 
                 447 
               
               
                   
                   
                 UGGUdTdT 
                   
                   
                 CAdTdT 
                   
                 AGCUGGU 
                   
               
               
                   
               
               
                 AD-69398 
                 A-139733 
                 gsusggacGfaGfAf 
                 270 
                 A-139734 
                 usUfsuugCfaGfAfgc 
                 359 
                 UGGUGGACGAGA 
                 448 
               
               
                   
                   
                 GfcucugcaaaaL96 
                   
                   
                 ucUfcGfuccacscsa 
                   
                 GCUCUGCAAAA 
                   
               
               
                   
               
               
                 AD-69443 
                 A-139822 
                 GGACGAGAGCUCUGC 
                 271 
                 A-139823 
                 UUUUGCAGAGCUCUCGU 
                 360 
                 GGACGAGAGCUC 
                 449 
               
               
                   
                   
                 AAAAdTdT 
                   
                   
                 CCdTdT 
                   
                 UGCAAAA 
                   
               
               
                   
               
               
                 AD-69380 
                 A-139697 
                 asasguacAfuGfGf 
                 272 
                 A-139698 
                 usAfsccaGfcUfCfgc 
                 361 
                 GCAAGUACAUGG 
                 450 
               
               
                   
                   
                 GfcgagcugguaL96 
                   
                   
                 ccAfuGfuacuusgsc 
                   
                 GCGAGCUGGUA 
                   
               
               
                   
               
               
                 AD-69425 
                 A-139786 
                 GUACAUGGGCGAGCU 
                 273 
                 A-139787 
                 UACCAGCUCGCCCAUGU 
                 362 
                 GUACAUGGGCGA 
                 451 
               
               
                   
                   
                 GGUAdTdT 
                   
                   
                 ACdTdT 
                   
                 GCUGGUA 
                   
               
               
                   
               
               
                 AD-69403 
                 A-139742 
                 asgsgcucGfuGfGf 
                 274 
                 A-139743 
                 usAfsgguUfuUfCfgu 
                 363 
                 UCAGGCUCGUGG 
                 452 
               
               
                   
                   
                 AfcgaaaaccuaL96 
                   
                   
                 ccAfcGfagccusgsa 
                   
                 ACGAAAACCUA 
                   
               
               
                   
               
               
                 AD-69447 
                 A-139830 
                 GCUCGUGGACGAAAA 
                 275 
                 A-139831 
                 UAGGUUUUCGUCCACGA 
                 364 
                 GCUCGUGGACGA 
                 453 
               
               
                   
                   
                 CCUAdTdT 
                   
                   
                 GCdTdT 
                   
                 AAACCUA 
                   
               
               
                   
               
               
                 AD-69404 
                 A-139744 
                 csgsuggaCfgAfAf 
                 276 
                 A-139745 
                 asAfsgagCfaGfGfuu 
                 365 
                 CUCGUGGACGAA 
                 454 
               
               
                   
                   
                 AfaccugcucuuL96 
                   
                   
                 uuCfgUfccacgsasg 
                   
                 AACCUGCUCUU 
                   
               
               
                   
               
               
                 AD-69405 
                 A-139746 
                 gsusggacGfaAfAf 
                 277 
                 A-139747 
                 usAfsagaGfcAfGfgu 
                 366 
                 UCGUGGACGAAA 
                 455 
               
               
                   
                   
                 AfccugcucuuaL96 
                   
                   
                 uuUfcGfuccacsgsa 
                   
                 ACCUGCUCUUA 
                   
               
               
                   
               
               
                 AD-69448 
                 A-139832 
                 UGGACGAAAACCUGC 
                 278 
                 A-139833 
                 AAGAGCAGGUUUUCGUC 
                 367 
                 UGGACGAAAACC 
                 456 
               
               
                   
                   
                 UCUUdTdT 
                   
                   
                 CAdTdT 
                   
                 UGCUCUU 
                   
               
               
                   
               
               
                 AD-69449 
                 A-139834 
                 GGACGAAAACCUGCU 
                 279 
                 A-139835 
                 UAAGAGGAGGUUUUCGU 
                 368 
                 GGACGAAAACCU 
                 457 
               
               
                   
                   
                 CUUAdTdT 
                   
                   
                 CCdTdT 
                   
                 GCUCUUA 
                   
               
               
                   
               
               
                 AD-69406 
                 A-139748 
                 csgscaagCfaGfAf 
                 280 
                 A-139749 
                 usAfsuguUfgUfAfga 
                 369 
                 ACCGCAAGCAGA 
                 458 
               
               
                   
                   
                 UfcuacaacauaL96 
                   
                   
                 ucUfgCfuugcgsgsu 
                   
                 UCUACAACAUA 
                   
               
               
                   
               
               
                 AD-69450 
                 A-139836 
                 CAAGCAGAUCUACAA 
                 281 
                 A-139837 
                 UAUGUUGUAGAUCUGCU 
                 370 
                 CAAGCAGAUCUA 
                 459 
               
               
                   
                   
                 CAUAdTdT 
                   
                   
                 UGdTdT 
                   
                 CAACAUA 
                   
               
               
                   
               
               
                 AD-69390 
                 A-139717 
                 asgscugcGfaGfAf 
                 282 
                 A-139718 
                 usAfsugaAfgGftfga 
                 371 
                 CCAGCUGCGAGA 
                 460 
               
               
                   
                   
                 UfcaccuucauaL96 
                   
                   
                 ncUfcGfcagcusgsg 
                   
                 UCACCUUCAUA 
                   
               
               
                   
               
               
                 AD-69435 
                 A-139806 
                 CtGCGAGAUCACCtU 
                 283 
                 A-139807 
                 UAUGAAGGUGAUCUCGC 
                 372 
                 CUGCGAGAUCAC 
                 461 
               
               
                   
                   
                 CAUAdTdT 
                   
                   
                 AGdTdT 
                   
                 CUUCAUA 
                   
               
               
                   
               
               
                 AD-69408 
                 A-139752 
                 cscsagucCfuGfGf 
                 284 
                 A-139753 
                 usAfsagaAfaAfUfgg 
                 373 
                 ACCCAGUCCUGG 
                 462 
               
               
                   
                   
                 CfcauuuuucuuaL96 
                   
                   
                 ccAfgGfacuggsgsu 
                   
                 CCAUUUUCUUA 
                   
               
               
                   
               
               
                 AD-69452 
                 A-139840 
                 AGUCCUGGCCAUUUUC 
                 285 
                 A-139841 
                 UAAGAAAAUGGCCAGGA 
                 374 
                 AGUCCUGGCCAU 
                 463 
               
               
                   
                   
                 UUAdTdT 
                   
                   
                 CUdTdT 
                   
                 UUUCUUA 
                   
               
               
                   
               
               
                 AD-69409 
                 A-139754 
                 csascugaGfuGfGfC 
                 286 
                 A-139755 
                 asGfsaauCfaCfAfag 
                 375 
                 AUCACUGAGUGG 
                 464 
               
               
                   
                   
                 fuugugauucuL96 
                   
                   
                 ccAfcUfcagugsasu 
                   
                 CUUGUGAUUCU 
                   
               
               
                   
               
               
                 AD-69453 
                 A-139842 
                 CUGAGUGGCUUGUGAU 
                 287 
                 A-139843 
                 AGAAUCACAAGCCACUC 
                 376 
                 CUGAGUGGCUUG 
                 465 
               
               
                   
                   
                 UCUdTdT 
                   
                   
                 AGdTdT 
                   
                 UGAUUCU 
                   
               
               
                   
               
               
                 AD-69410 
                 A-139756 
                 asasuguuAfaAfAfG 
                 288 
                 A-139757 
                 asUfsguuUfaAfAfac 
                 377 
                 AAAAUGUUAAAA 
                 466 
               
               
                   
                   
                 fuuuuaaacauL96 
                   
                   
                 uuUfuAfacauususu 
                   
                 GUUUUAAACAU 
                   
               
               
                   
               
               
                 AD-69454 
                 A-139844 
                 UGUUAAAAGUUUUAAA 
                 289 
                 A-139845 
                 AUGUUUAAAACUUUUAA 
                 378 
                 UGUUAAAAGUUU 
                 467 
               
               
                   
                   
                 CAUdTdT 
                   
                   
                 CAdTdT 
                   
                 UAAACAU 
                   
               
               
                   
               
               
                 AD-69444 
                 A-139824 
                 AGCAGAAGGGAACAAC 
                 290 
                 A-139825 
                 UAUGUUGUUCCCUUCUG 
                 379 
                 AGCAGAAGGGAA 
                 468 
               
               
                   
                   
                 AUAdTdT 
                   
                   
                 CUdTdT 
                   
                 CAACAUA 
                   
               
               
                   
               
               
                 AD-69399 
                 A-139735 
                 gsgsagcaGfaAfGfG 
                 291 
                 A-139736 
                 usAfsuguUfgUfUfcc 
                 380 
                 CCGGAGCAGAAG 
                 469 
               
               
                   
                   
                 fgaacaacauaL96 
                   
                   
                 cuUfcUfgcuccsgsg 
                   
                 GGAACAACAUA 
                   
               
               
                   
               
               
                 AD-69445 
                 A-139826 
                 UCUCCGAGAUGCUAUC 
                 292 
                 A-139827 
                 UUUGAUAGCAUCUCGGA 
                 381 
                 UCUCCGAGAUGC 
                 470 
               
               
                   
                   
                 AAAdTdT 
                   
                   
                 GAdTdT 
                   
                 UAUCAAA 
                   
               
               
                   
               
               
                 AD-69400 
                 A-139737 
                 csusucucCfgAfCfA 
                 293 
                 A-139738 
                 usUfsugaUfaGfCfau 
                 382 
                 GACUUCUCCGAG 
                 471 
               
               
                   
                   
                 fugcuaucaaaL96 
                   
                   
                 cuCfgGfagaagsusc 
                   
                 AUGCUAUCAAA 
                   
               
               
                   
               
               
                 AD-69446 
                 A-139828 
                 AGAUGGAUGUGGUGGC 
                 294 
                 A-139829 
                 AUUGCCACCACAUCCAU 
                 383 
                 AGAUGGAUGUGG 
                 472 
               
               
                   
                   
                 AAUdTdT 
                   
                   
                 CUdTdT 
                   
                 UGGCAAU 
                   
               
               
                   
               
               
                 AD-69401 
                 A-139739 
                 usgsagauGfgAfUfG 
                 295 
                 A-139740 
                 asUfsugcCfaCfCfac 
                 384 
                 UUUGAGAUGGAU 
                 473 
               
               
                   
                   
                 fugguggcaauL96 
                   
                   
                 auCfcAfucucasasa 
                   
                 GUGGUGGCAAU 
                   
               
               
                   
               
               
                 AD-69451 
                 A-139838 
                 CUGCGAAAUCACCUUC 
                 296 
                 A-139839 
                 AAUGAAGGUGAUUUCGC 
                 385 
                 CUGCGAAAUCAC 
                 474 
               
               
                   
                   
                 AUUdTdT 
                   
                   
                 AGdTdT 
                   
                 CUUCAUU 
                   
               
               
                   
               
               
                 AD-69407 
                 A-139750 
                 asascugcGfaAfAfU 
                 297 
                 A-139751 
                 asAfsuguAfgGfUfga 
                 386 
                 CCAACUGCGAAA 
                 475 
               
               
                   
                   
                 fcaccuucauuL96 
                   
                   
                 uuUfcGfcaguusgsg 
                   
                 UCACCUUCAUU 
               
               
                   
               
            
           
         
       
     
     
       
         
           
               
             
               
                 TABLE 4 
               
             
            
               
                   
               
               
                 GCK Single Dose Screen in Primary Mouse Hepatocytes 
               
            
           
           
               
               
            
               
                   
                 Primary Mouse Hepatocytes 
               
            
           
           
               
               
               
               
               
            
               
                 Duplex Name 
                 10 nM Avg 
                 10 nM SD 
                 0.1 nM Avg 
                 0.1 nM SD 
               
               
                   
               
            
           
           
               
               
               
               
               
            
               
                 AD-69366 
                 82.2 
                 54.1 
                 70.2 
                 12.8 
               
               
                 AD-69368 
                 47.8 
                 5.3 
                 91.7 
                 3.6 
               
               
                 AD-69411 
                 17.6 
                 4.8 
                 27.9 
                 2.8 
               
               
                 AD-69413 
                 15.9 
                 5.1 
                 43.7 
                 4.8 
               
               
                 AD-69367 
                 58.2 
                 11.0 
                 92.0 
                 10.5 
               
               
                 AD-69369 
                 60.3 
                 10.6 
                 88.0 
                 6.5 
               
               
                 AD-69412 
                 24.1 
                 5.3 
                 71.7 
                 15.6 
               
               
                 AD-69414 
                 52.5 
                 7.7 
                 82.7 
                 7.2 
               
               
                 AD-69371 
                 77.5 
                 9.7 
                 89.1 
                 12.3 
               
               
                 AD-69416 
                 11.8 
                 2.4 
                 25.1 
                 4.0 
               
               
                 AD-69370 
                 99.5 
                 10.1 
                 96.9 
                 5.1 
               
               
                 AD-69415 
                 90.3 
                 9.3 
                 91.3 
                 15.2 
               
               
                 AD-69372 
                 27.9 
                 2.2 
                 49.8 
                 10.2 
               
               
                 AD-69417 
                 29.9 
                 2.1 
                 63.6 
                 12.6 
               
               
                 AD-69373 
                 96.7 
                 17.1 
                 104.7 
                 48.8 
               
               
                 AD-69418 
                 86.2 
                 34.0 
                 62.0 
                 9.9 
               
               
                 AD-69374 
                 33.7 
                 4.8 
                 71.7 
                 7.8 
               
               
                 AD-69419 
                 17.1 
                 3.2 
                 44.8 
                 4.4 
               
               
                 AD-69375 
                 72.0 
                 8.3 
                 104.5 
                 5.3 
               
               
                 AD-69420 
                 20.3 
                 2.0 
                 46.6 
                 7.2 
               
               
                 AD-69376 
                 22.1 
                 2.3 
                 59.0 
                 9.7 
               
               
                 AD-69377 
                 23.0 
                 2.0 
                 59.2 
                 6.6 
               
               
                 AD-69421 
                 16.1 
                 2.1 
                 29.9 
                 3.5 
               
               
                 AD-69422 
                 15.2 
                 1.2 
                 38.9 
                 2.4 
               
               
                 AD-69378 
                 24.8 
                 2.1 
                 70.1 
                 4.2 
               
               
                 AD-69379 
                 11.2 
                 0.9 
                 18.0 
                 3.2 
               
               
                 AD-69423 
                 11.7 
                 1.3 
                 41.1 
                 3.6 
               
               
                 AD-69424 
                 9.5 
                 1.6 
                 13.9 
                 3.5 
               
               
                 AD-69381 
                 73.6 
                 13.3 
                 99.9 
                 36.4 
               
               
                 AD-69426 
                 84.5 
                 6.3 
                 76.5 
                 6.1 
               
               
                 AD-69382 
                 91.9 
                 5.6 
                 91.6 
                 3.3 
               
               
                 AD-69427 
                 97.6 
                 2.5 
                 88.8 
                 4.5 
               
               
                 AD-69383 
                 95.9 
                 3.2 
                 100.5 
                 9.3 
               
               
                 AD-69428 
                 32.7 
                 4.3 
                 66.1 
                 5.0 
               
               
                 AD-69384 
                 88.3 
                 9.8 
                 97.9 
                 8.6 
               
               
                 AD-69429 
                 90.7 
                 10.6 
                 95.9 
                 5.7 
               
               
                 AD-69385 
                 29.2 
                 4.2 
                 45.1 
                 4.4 
               
               
                 AD-69386 
                 92.5 
                 9.0 
                 97.8 
                 9.8 
               
               
                 AD-69430 
                 21.2 
                 1.4 
                 31.8 
                 2.1 
               
               
                 AD-69387 
                 69.7 
                 9.0 
                 77.8 
                 7.1 
               
               
                 AD-69432 
                 69.5 
                 3.2 
                 79.6 
                 13.6 
               
               
                 AD-69388 
                 66.9 
                 11.3 
                 84.7 
                 11.9 
               
               
                 AD-69389 
                 76.3 
                 18.3 
                 86.9 
                 6.0 
               
               
                 AD-69433 
                 70.0 
                 2.6 
                 76.4 
                 5.0 
               
               
                 AD-69434 
                 97.6 
                 6.6 
                 71.6 
                 9.5 
               
               
                 AD-69391 
                 21.7 
                 5.1 
                 59.8 
                 5.6 
               
               
                 AD-69436 
                 19.9 
                 1.9 
                 54.3 
                 14.6 
               
               
                 AD-69392 
                 39.2 
                 4.8 
                 81.5 
                 12.7 
               
               
                 AD-69402 
                 32.2 
                 5.1 
                 62.9 
                 12.4 
               
               
                 AD-69393 
                 34.9 
                 5.4 
                 78.5 
                 13.0 
               
               
                 AD-69437 
                 14.0 
                 2.5 
                 27.3 
                 4.6 
               
               
                 AD-69438 
                 11.7 
                 0.7 
                 28.4 
                 2.5 
               
               
                 AD-69395 
                 28.2 
                 6.8 
                 55.4 
                 12.2 
               
               
                 AD-69440 
                 19.4 
                 2.8 
                 34.7 
                 7.3 
               
               
                 AD-69396 
                 29.7 
                 3.0 
                 57.2 
                 9.8 
               
               
                 AD-69441 
                 23.8 
                 4.1 
                 44.6 
                 7.4 
               
               
                 AD-69397 
                 39.5 
                 11.8 
                 67.9 
                 11.6 
               
               
                 AD-69442 
                 29.3 
                 2.4 
                 40.1 
                 8.4 
               
               
                 AD-69394 
                 39.4 
                 4.2 
                 79.5 
                 3.2 
               
               
                 AD-69439 
                 37.6 
                 3.2 
                 65.9 
                 3.0 
               
               
                 AD-69398 
                 92.2 
                 9.6 
                 111.3 
                 16.2 
               
               
                 AD-69443 
                 115.1 
                 14.5 
                 97.4 
                 5.1 
               
               
                 AD-69380 
                 23.6 
                 3.2 
                 65.0 
                 11.2 
               
               
                 AD-69425 
                 11.9 
                 1.2 
                 29.4 
                 6.5 
               
               
                 AD-69403 
                 64.2 
                 13.0 
                 77.0 
                 13.9 
               
               
                 AD-69447 
                 87.4 
                 7.9 
                 84.3 
                 11.7 
               
               
                 AD-69404 
                 67.2 
                 10.6 
                 83.9 
                 16.6 
               
               
                 AD-69405 
                 69.0 
                 10.9 
                 86.1 
                 23.6 
               
               
                 AD-69448 
                 107.8 
                 6.4 
                 120.4 
                 36.9 
               
               
                 AD-69449 
                 129.6 
                 37.4 
                 120.2 
                 6.4 
               
               
                 AD-69406 
                 63.8 
                 4.7 
                 91.7 
                 10.8 
               
               
                 AD-69450 
                 84.5 
                 12.4 
                 109.5 
                 5.3 
               
               
                 AD-69390 
                 16.9 
                 4.2 
                 38.9 
                 11.6 
               
               
                 AD-69435 
                 9.0 
                 1.5 
                 20.9 
                 3.5 
               
               
                 AD-69408 
                 67.3 
                 4.7 
                 83.6 
                 16.3 
               
               
                 AD-69452 
                 92.6 
                 10.4 
                 111.8 
                 5.1 
               
               
                 AD-69409 
                 74.8 
                 4.8 
                 79.7 
                 12.3 
               
               
                 AD-69453 
                 87.2 
                 1.4 
                 93.7 
                 3.0 
               
               
                 AD-69410 
                 71.2 
                 4.9 
                 90.2 
                 9.4 
               
               
                 AD-69454 
                 90.0 
                 10.9 
                 86.0 
                 7.8 
               
               
                 AD-69399 
                 24.4 
                 2.5 
                 45.8 
                 6.1 
               
               
                 AD-69445 
                 16.0 
                 0.6 
                 44.4 
                 4.4 
               
               
                 AD-69400 
                 12.2 
                 1.4 
                 34.8 
                 2.3 
               
               
                 AD-69446 
                 14.9 
                 1.3 
                 30.4 
                 2.9 
               
               
                 AD-69401 
                 15.4 
                 1.6 
                 29.4 
                 4.9 
               
               
                 AD-69451 
                 6.9 
                 1.3 
                 19.2 
                 3.6 
               
               
                 AD-69407 
                 4.1 
                 0.4 
                 11.8 
                 2.8 
               
               
                 AD-1955 
                 101.283 
                 16.1184 
               
               
                   
               
            
           
         
       
     
     
       
         
           
               
             
               
                 TABLE 5 
               
             
            
               
                   
               
               
                 GCK Single Dose Screen in Primary Cynomologous Hepatocytes 
               
            
           
           
               
               
            
               
                   
                 Primary Cyno Hepatocytes 
               
            
           
           
               
               
               
               
               
            
               
                 duplexName 
                 10 nM Avg 
                 10 nM SD 
                 0.1 nM Avg 
                 0.1 nM SD 
               
               
                   
               
            
           
           
               
               
               
               
               
            
               
                 AD-69366 
                 29.8 
                 14.3 
                 37.4 
                 9.3 
               
               
                 AD-69368 
                 12.2 
                 4.6 
                 28.1 
                 4.9 
               
               
                 AD-69411 
                 14.2 
                 4.5 
                 22.6 
                 10.8 
               
               
                 AD-69413 
                 12.1 
                 3.2 
                 30.0 
                 14.0 
               
               
                 AD-69367 
                 14.9 
                 2.6 
                 39.9 
                 7.1 
               
               
                 AD-69369 
                 36.6 
                 14.2 
                 54.2 
                 9.5 
               
               
                 AD-69412 
                 19.3 
                 6.5 
                 31.2 
                 4.6 
               
               
                 AD-69414 
                 26.4 
                 11.9 
                 63.6 
                 8.7 
               
               
                 AD-69371 
                 50.2 
                 20.5 
                 91.3 
                 17.3 
               
               
                 AD-69416 
                 24.5 
                 8.3 
                 21.8 
                 3.6 
               
               
                 AD-69370 
                 43.4 
                 10.3 
                 69.4 
                 11.6 
               
               
                 AD-69415 
                 15.4 
                 6.2 
                 34.2 
                 13.2 
               
               
                 AD-69372 
                 19.8 
                 7.9 
                 23.7 
                 8.7 
               
               
                 AD-69417 
                 23.8 
                 10.5 
                 31.4 
                 16.1 
               
               
                 AD-69373 
                 87.7 
                 22.1 
                 66.8 
                 10.6 
               
               
                 AD-69418 
                 20.4 
                 4.9 
                 25.7 
                 6.4 
               
               
                 AD-69374 
                 24.7 
                 15.0 
                 41.8 
                 6.2 
               
               
                 AD-69419 
                 21.9 
                 6.7 
                 32.8 
                 12.9 
               
               
                 AD-69375 
                 20.6 
                 5.5 
                 49.5 
                 16.0 
               
               
                 AD-69420 
                 15.3 
                 2.9 
                 41.8 
                 21.6 
               
               
                 AD-69376 
                 17.7 
                 5.3 
                 22.4 
                 12.1 
               
               
                 AD-69377 
                 21.6 
                 5.6 
                 26.4 
                 6.2 
               
               
                 AD-69421 
                 22.1 
                 4.7 
                 22.1 
                 8.9 
               
               
                 AD-69422 
                 22.7 
                 5.7 
                 38.2 
                 11.8 
               
               
                 AD-69378 
                 14.7 
                 6.6 
                 31.9 
                 26.2 
               
               
                 AD-69379 
                 10.5 
                 2.7 
                 14.4 
                 2.9 
               
               
                 AD-69423 
                 20.3 
                 8.2 
                 42.2 
                 10.8 
               
               
                 AD-69424 
                 20.0 
                 5.1 
                 17.5 
                 10.3 
               
               
                 AD-69381 
                 46.6 
                 12.8 
                 56.9 
                 20.6 
               
               
                 AD-69426 
                 49.3 
                 8.4 
                 76.7 
                 29.0 
               
               
                 AD-69382 
                 74.0 
                 22.7 
                 54.6 
                 14.2 
               
               
                 AD-69427 
                 33.9 
                 5.9 
                 66.3 
                 13.8 
               
               
                 AD-69383 
                 63.4 
                 18.6 
                 81.4 
                 22.7 
               
               
                 AD-69428 
                 39.4 
                 16.0 
                 69.7 
                 10.0 
               
               
                 AD-69384 
                 58.1 
                 17.4 
                 89.1 
                 22.8 
               
               
                 AD-69429 
                 30.9 
                 7.3 
                 47.9 
                 17.9 
               
               
                 AD-69385 
                 27.3 
                 9.2 
                 37.0 
                 7.7 
               
               
                 AD-69386 
                 41.4 
                 16.6 
                 61.3 
                 18.6 
               
               
                 AD-69430 
                 24.2 
                 8.8 
                 29.8 
                 13.7 
               
               
                 AD-69387 
                 28.2 
                 3.0 
                 30.9 
                 11.1 
               
               
                 AD-69432 
                 23.5 
                 7.8 
                 24.4 
                 5.3 
               
               
                 AD-69388 
                 63.7 
                 11.0 
                 70.8 
                 7.3 
               
               
                 AD-69389 
                 59.7 
                 13.8 
                 77.8 
                 15.6 
               
               
                 AD-69433 
                 37.0 
                 3.3 
                 62.2 
                 31.0 
               
               
                 AD-69434 
                 76.3 
                 27.9 
                 94.1 
                 8.0 
               
               
                 AD-69391 
                 22.0 
                 11.2 
                 25.2 
                 10.0 
               
               
                 AD-69436 
                 24.5 
                 4.8 
                 41.3 
                 2.7 
               
               
                 AD-69392 
                 23.3 
                 10.1 
                 52.9 
                 11.4 
               
               
                 AD-69402 
                 21.9 
                 7.6 
                 59.1 
                 29.2 
               
               
                 AD-69393 
                 22.9 
                 2.3 
                 37.3 
                 2.9 
               
               
                 AD-69437 
                 21.8 
                 6.6 
                 30.4 
                 7.7 
               
               
                 AD-69438 
                 21.5 
                 5.9 
                 32.4 
                 9.6 
               
               
                 AD-69395 
                 17.4 
                 5.7 
                 32.3 
                 4.6 
               
               
                 AD-69440 
                 25.7 
                 15.0 
                 35.1 
                 13.5 
               
               
                 AD-69396 
                 28.8 
                 9.3 
                 16.6 
                 5.1 
               
               
                 AD-69441 
                 26.3 
                 8.4 
                 39.2 
                 8.9 
               
               
                 AD-69397 
                 62.4 
                 33.3 
                 48.6 
                 26.1 
               
               
                 AD-69442 
                 41.0 
                 12.3 
                 50.4 
                 12.5 
               
               
                 AD-69394 
                 38.0 
                 12.9 
                 65.7 
                 13.1 
               
               
                 AD-69439 
                 30.2 
                 9.0 
                 51.6 
                 10.8 
               
               
                 AD-69398 
                 21.1 
                 3.8 
                 23.1 
                 6.5 
               
               
                 AD-69443 
                 20.0 
                 7.6 
                 29.3 
                 4.2 
               
               
                 AD-69380 
                 43.8 
                 15.3 
                 64.7 
                 26.0 
               
               
                 AD-69425 
                 35.5 
                 9.8 
                 38.5 
                 8.5 
               
               
                 AD-69403 
                 34.5 
                 12.0 
                 35.1 
                 7.0 
               
               
                 AD-69447 
                 23.5 
                 7.4 
                 55.0 
                 21.7 
               
               
                 AD-69404 
                 41.2 
                 23.9 
                 30.0 
                 5.5 
               
               
                 AD-69405 
                 20.2 
                 7.7 
                 35.0 
                 14.4 
               
               
                 AD-69448 
                 53.1 
                 16.4 
                 42.3 
                 6.3 
               
               
                 AD-69449 
                 37.8 
                 14.7 
                 42.6 
                 4.0 
               
               
                 AD-69406 
                 21.6 
                 13.7 
                 23.4 
                 7.1 
               
               
                 AD-69450 
                 19.6 
                 4.5 
                 23.9 
                 5.1 
               
               
                 AD-69390 
                 18.8 
                 8.0 
                 21.0 
                 4.2 
               
               
                 AD-69435 
                 20.0 
                 17.3 
                 23.9 
                 9.7 
               
               
                 AD-69408 
                 40.0 
                 17.9 
                 34.1 
                 18.5 
               
               
                 AD-69452 
                 28.9 
                 9.6 
                 45.5 
                 13.6 
               
               
                 AD-69409 
                 40.0 
                 10.8 
                 52.7 
                 31.5 
               
               
                 AD-69453 
                 39.2 
                 6.5 
                 57.6 
                 20.9 
               
               
                 AD-69410 
                 62.0 
                 8.7 
                 78.6 
                 10.5 
               
               
                 AD-69454 
                 86.5 
                 6.9 
                 102.4 
                 27.8 
               
               
                 AD-69399 
                 88.8 
                 17.5 
                 81.6 
                 21.4 
               
               
                 AD-69445 
                 68.9 
                 19.5 
                 118.3 
                 26.5 
               
               
                 AD-69400 
                 32.7 
                 11.9 
                 31.1 
                 10.1 
               
               
                 AD-69446 
                 23.5 
                 10.8 
                 38.3 
                 4.1 
               
               
                 AD-69401 
                 28.4 
                 18.3 
                 24.7 
                 5.7 
               
               
                 AD-69451 
                 79.6 
                 23.5 
                 102.9 
                 41.6 
               
               
                 AD-69407 
                 16.8 
                 7.7 
                 29.2 
                 12.6 
               
               
                 AD-1955 
                 102.912 
                 29.2078 
               
               
                   
               
            
           
         
       
     
     Example 2. iRNA Design, Synthesis, Selection, and In Vitro Evaluation 
     This Example describes methods for the design, synthesis, selection, and in vitro evaluation of additional GCK iRNA agents. 
     Bioinformatics 
     A set of siRNAs targeting human glucokinase (GCK) (human NCBI refseqID: NM_033507; NCBI GeneID: 2645) were designed using custom R and Python scripts. The human GCK REFSEQ mRNA has a length of 2442 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 position 2442 was determined with a linear model derived the direct measure of mRNA knockdown from more than 20,000 distinct siRNA designs targeting a large number of vertebrate genes. The custom Python script built the set of siRNAs by systematically selecting an siRNA every 11 bases along the target mRNA nucleotide seqeunce starting at position 10. At each of the positions, the neighboring siRNA (one position to the 5′ end of the mRNA, one position to the 3′ end of the mRNA) was swapped into the design set if the predicted efficacy was better than the efficacy at the exact every-eleventh siRNA. Low complexity siRNAs, e.g., those with Shannon Entropy measures below 1.35, were excluded from the set. 
     Cell Culture and Transfections 
     Primary Cyno Hepatocytes (PCH) cells were transfected by adding 4.9 μl of Opti-MEM plus 0.1 μl of Lipofectamine RNAiMax per well (Invitrogen, Carlsbad Calif. cat #13778-150) to 5 μl of siRNA duplexes per well into a 384-well plate and incubated at room temperature for 15 minutes. Forty μl of EMEM containing about 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 20 nM final duplex concentration. 
     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, 50 μl of Lysis/Binding Buffer and 25 μl of lysis buffer containing 3 μl 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 two times with 150 μl Wash Buffer A and once with Wash Buffer B. Beads were then washed with 150 μl Elution Buffer, re-captured, and the supernatant was removed. 
     cDNA Synthesis Using ABI High Capacity cDNA Reverse Transcription Kit (Applied Biosystems, Foster City, Calif., Cat #4368813) 
     Ten μl of a master mix containing 1 μl 10× Buffer, 0.4 μl 125× dNTPs, 1 μl 10× Random primers, 0.5 μl Reverse Transcriptase, 0.5 μl RNase inhibitor and 6.6 μl of H2O per reaction was added to RNA isolated as described above. Plates were sealed, mixed, and incubated on an electromagnetic shaker for 10 minutes at room temperature, followed by 2 hours at 37° C. 
     Real Time PCR 
     Two μl of cDNA were added to a master mix containing 0.5 μl of Custom Cyno GAPDH TaqMan Probe, 0.5 μl cyno GCK probe (Mf02827184 ml) and 5 μl Lightcycler 480 probe master mix (Roche Cat #04887301001) per well in a 384 well plates (Roche cat #04887301001). Real time PCR was performed in a LightCycler480 Real Time PCR system (Roche) using the ΔΔCt(RQ) assay. Each duplex was tested in four independent transfections. 
     To calculate relative fold change, real time data were analyzed using the ΔΔCt method and normalized to assays performed with cells transfected with 20 nM AD-1955, or mock transfected cells. 
     A detailed list of the unmodified GCK sense and antisense strand sequences is shown in Table 6 and a detailed list of the modified GCK sense and antisense strand sequences is shown in Table 7. 
     Table 8 shows the results of a single dose screen in primary Cynomolgus hepatocytes (PCH) transfected with the indicated modified siRNAs. Data are expressed as percent of message remaining relative to cells treated with a non-targeting control siRNA, AD-1955. 
     
       
         
           
               
             
               
                 TABLE 6 
               
             
            
               
                   
               
               
                 GCK Unmodified Sequences 
               
            
           
           
               
               
               
               
               
               
               
               
            
               
                   
                   
                   
                   
                 Anti- 
                   
                   
                   
               
               
                   
                 Sense 
                   
                 SEQ 
                 ssense 
                   
                 SEQ 
                   
               
               
                 Duplex 
                 Oligo 
                 Sense Sequence 
                 ID 
                 Oligo 
                 Sense Sequence 
                 ID 
                 Position in 
               
               
                 Name 
                 Name 
                 (5′ to 3′) 
                 NO: 
                 Name 
                 (5′ to 3′) 
                 NO: 
                 NM_033507.1 
               
               
                   
               
               
                 AD-71009 
                 A-142377 
                 CUGCCAGCCUCAGGCAGCU 
                 476 
                 A-142378 
                 AGCUGCCUGAGGCUGGCAG 
                 684 
                 24-42 
               
               
                   
               
               
                 AD-71010 
                 A-142379 
                 UCAGGCAGCUCUCCAUCCA 
                 477 
                 A-142380 
                 UGGAUGGAGAGCUGCCUGA 
                 685 
                 33-51 
               
               
                   
               
               
                 AD-71011 
                 A-142381 
                 CCAUCCAAGCAGCCGUUGA 
                 478 
                 A-142382 
                 UCAACGGCUGCUUGGAUGG 
                 686 
                 45-63 
               
               
                   
               
               
                 AD-71012 
                 A-142383 
                 AGCCGUUGCUGCCACAGGA 
                 479 
                 A-142384 
                 UCCUGUGGCAGCAACGGCU 
                 687 
                 55-73 
               
               
                   
               
               
                 AD-71013 
                 A-142385 
                 ACAGGCGGGCCUUACGCUA 
                 480 
                 A-142386 
                 UAGCGUAAGGCCCGCCUGU 
                 688 
                 68-86 
               
               
                   
               
               
                 AD-71014 
                 A-142387 
                 UUACGCUCCAAGGCUACAA 
                 481 
                 A-142388 
                 UUGUAGCCUUGGAGCGUAA 
                 689 
                 79-97 
               
               
                   
               
               
                 AD-71015 
                 A-142389 
                 AAGGCUACAGCAUGUGCUA 
                 482 
                 A-142390 
                 UAGCACAUGCUGUAGCCUU 
                 690 
                  88-106 
               
               
                   
               
               
                 AD-71016 
                 A-142391 
                 UGUGCUAGGCCUCAGCAGA 
                 483 
                 A-142392 
                 UCUGCUGAGGCCUAGCACA 
                 691 
                 100-118 
               
               
                   
               
               
                 AD-71017 
                 A-142393 
                 UCAGCAGGCAGGAGCAUCU 
                 484 
                 A-142394 
                 AGAUGCUCCUGCCUGCUGA 
                 692 
                 111-129 
               
               
                   
               
               
                 AD-71018 
                 A-142395 
                 AGCAUCUCUGCCUCCCAAA 
                 485 
                 A-142396 
                 UUUGGGAGGCAGAGAUGCU 
                 693 
                 123-141 
               
               
                   
               
               
                 AD-71019 
                 A-142397 
                 CCUCCCAAAGCAUCUACCU 
                 486 
                 A-142398 
                 AGGUAGAUGCUUUGGGAGG 
                 694 
                 133-151 
               
               
                   
               
               
                 AD-71020 
                 A-142401 
                 UAGCCCCUCGGAGAGAUGA 
                 487 
                 A-142402 
                 UCAUCUCUCCGAGGGGCUA 
                 695 
                 154-172 
               
               
                   
               
               
                 AD-71021 
                 A-142403 
                 AGAGAUGGCGAUGGAUGUA 
                 488 
                 A-142404 
                 UACAUCCAUCGCCAUCUCU 
                 696 
                 165-183 
               
               
                   
               
               
                 AD-71022 
                 A-142405 
                 UGGAUGUCACAAGGAGCCA 
                 489 
                 A-142406 
                 UGGCUCCUUGUGACAUCCA 
                 697 
                 176-194 
               
               
                   
               
               
                 AD-71023 
                 A-142407 
                 AGGAGCCAGGCCCAGACAA 
                 490 
                 A-142408 
                 UUGUCUGGGCCUGGCUCCU 
                 698 
                 187-205 
               
               
                   
               
               
                 AD-71024 
                 A-142411 
                 ACUCUGGUAGAGCAGAUCA 
                 491 
                 A-142412 
                 UGAUCUGCUCUACCAGAGU 
                 699 
                 211-229 
               
               
                   
               
               
                 AD-71025 
                 A-142413 
                 AGCAGAUCCUGGCAGAGUU 
                 492 
                 A-142414 
                 AACUCUGCCAGGAUCUGCU 
                 700 
                 221-239 
               
               
                   
               
               
                 AD-71026 
                 A-142415 
                 CAGAGUUCCAGCUGCAGGA 
                 493 
                 A-142416 
                 UCCUGCAGCUGGAACUCUG 
                 701 
                 233-251 
               
               
                   
               
               
                 AD-71027 
                 A-142417 
                 AGCUGCAGGAGGAGGACCU 
                 494 
                 A-142418 
                 AGGUCCUCCUCCUGCAGCU 
                 702 
                 242-260 
               
               
                   
               
               
                 AD-71028 
                 A-142419 
                 AGGACCUGAAGAAGGUGAU 
                 495 
                 A-142420 
                 AUCACCUUCUUCAGGUCCU 
                 703 
                 254-272 
               
               
                   
               
               
                 AD-71029 
                 A-142421 
                 AAGGUGAUGAGACGGAUGA 
                 496 
                 A-142422 
                 UCAUCCGUCUCAUCACCUU 
                 704 
                 265-283 
               
               
                   
               
               
                 AD-71030 
                 A-142423 
                 CGGAUGCAGAAGGAGAUGA 
                 497 
                 A-142424 
                 UCAUCUCCUUCUGCAUCCG 
                 705 
                 277-295 
               
               
                   
               
               
                 AD-71031 
                 A-142425 
                 AAGGAGAUGGACCGCGGCA 
                 498 
                 A-142426 
                 UGCCGCGGUCCAUCUCCUU 
                 706 
                 286-304 
               
               
                   
               
               
                 AD-71032 
                 A-142427 
                 CGCGGCCUGAGGCUGGAGA 
                 499 
                 A-142428 
                 UCUCCAGCCUCAGGCCGCG 
                 707 
                 298-316 
               
               
                   
               
               
                 AD-71033 
                 A-142429 
                 CUGGAGACCCAUGAAGAGA 
                 500 
                 A-142430 
                 UCUCUUCAUGGGUCUCCAG 
                 708 
                 310-328 
               
               
                   
               
               
                 AD-71034 
                 A-142431 
                 CAUGAAGAGGCCAGUGUGA 
                 501 
                 A-142432 
                 UCACACUGGCCUCUUCAUG 
                 709 
                 319-337 
               
               
                   
               
               
                 AD-71035 
                 A-142433 
                 CAGUGUGAAGAUGCUGCCA 
                 502 
                 A-142434 
                 UGGCAGCAUCUUCACACUG 
                 710 
                 330-348 
               
               
                   
               
               
                 AD-71036 
                 A-142435 
                 UGCUGCCCACCUACGUGCA 
                 503 
                 A-142436 
                 UGCACGUAGGUGGGCAGCA 
                 711 
                 341-359 
               
               
                   
               
               
                 AD-71037 
                 A-142437 
                 UACGUGCGCUCCACCCCAA 
                 504 
                 A-142438 
                 UUGGGGUGGAGCGCACGUA 
                 712 
                 352-370 
               
               
                   
               
               
                 AD-71038 
                 A-142439 
                 ACCCCAGAAGGCUCAGAAA 
                 505 
                 A-142440 
                 UUUCUGAGCCUUCUGGGGU 
                 713 
                 364-382 
               
               
                   
               
               
                 AD-71039 
                 A-142441 
                 UCAGAAGUCGGGGACUUCA 
                 506 
                 A-142442 
                 UGAAGUCCCCGACUUCUGA 
                 714 
                 376-394 
               
               
                   
               
               
                 AD-71040 
                 A-142443 
                 GGGGACUUCCUCUCCCUGA 
                 507 
                 A-142444 
                 UCAGGGAGAGGAAGUCCCC 
                 715 
                 385-403 
               
               
                   
               
               
                 AD-71041 
                 A-142445 
                 UCCCUGGACCUGGGUGGCA 
                 508 
                 A-142446 
                 UGCCACCCAGGUCCAGGGA 
                 716 
                 397-415 
               
               
                   
               
               
                 AD-71042 
                 A-142447 
                 UGGGUGGCACUAACUUCAA 
                 509 
                 A-142448 
                 UUGAAGUUAGUGCCACCCA 
                 717 
                 407-425 
               
               
                   
               
               
                 AD-71043 
                 A-142449 
                 ACUUCAGGGUGAUGCUGGU 
                 510 
                 A-142450 
                 ACCAGCAUCACCCUGAAGU 
                 718 
                 419-437 
               
               
                   
               
               
                 AD-71044 
                 A-142453 
                 AGGUGGGAGAAGGUGAGGA 
                 511 
                 A-142454 
                 UCCUCACCUUCUCCCACCU 
                 719 
                 440-458 
               
               
                   
               
               
                 AD-71045 
                 A-142457 
                 CAGUGGAGCGUGAAGACCA 
                 512 
                 A-142458 
                 UGGUCUUCACGCUCCACUG 
                 720 
                 463-481 
               
               
                   
               
               
                 AD-71046 
                 A-142461 
                 CCAGAUGUACUCCAUCCCA 
                 513 
                 A-142462 
                 UGGGAUGGAGUACAUCUGG 
                 721 
                 486-504 
               
               
                   
               
               
                 AD-71047 
                 A-142467 
                 ACCGGCACUGCUGAGAUGA 
                 514 
                 A-142468 
                 UCAUCUCAGCAGUGCCGGU 
                 722 
                 517-535 
               
               
                   
               
               
                 AD-71048 
                 A-142469 
                 AGAUGCUCUUCGACUACAU 
                 515 
                 A-142470 
                 AUGUAGUCGAAGAGCAUCU 
                 723 
                 530-548 
               
               
                   
               
               
                 AD-71049 
                 A-142471 
                 UCGACUACAUCUCUGAGUA 
                 516 
                 A-142472 
                 UACUCAGAGAUGUAGUCGA 
                 724 
                 539-557 
               
               
                   
               
               
                 AD-71050 
                 A-142473 
                 UCUGAGUGCAUCUCCGACU 
                 517 
                 A-142474 
                 AGUCGGAGAUGCACUCAGA 
                 725 
                 550-568 
               
               
                   
               
               
                 AD-71051 
                 A-142475 
                 UCCGACUUCCUGGACAAGA 
                 518 
                 A-142476 
                 UCUUGUCCAGGAAGUCGGA 
                 726 
                 562-580 
               
               
                   
               
               
                 AD-71052 
                 A-142477 
                 GACAAGCAUCAGAUGAAAC 
                 519 
                 A-142478 
                 GUUUCAUCUGAUGCUUGUC 
                 727 
                 574-592 
               
               
                   
               
               
                 AD-71053 
                 A-142479 
                 AGAUGAAACACAAGAAGCU 
                 520 
                 A-142480 
                 AGCUUCUUGUGUUUCAUCU 
                 728 
                 584-602 
               
               
                   
               
               
                 AD-71054 
                 A-142481 
                 AGAAGCUGCCCCUGGGCUU 
                 521 
                 A-142482 
                 AAGCCCAGGGGCAGCUUCU 
                 729 
                 596-614 
               
               
                   
               
               
                 AD-71055 
                 A-142483 
                 CCUGGGCUUCACCUUCUCA 
                 522 
                 A-142484 
                 UGAGAAGGUGAAGCCCAGG 
                 730 
                 606-624 
               
               
                   
               
               
                 AD-71056 
                 A-142485 
                 ACCUUCUCCUUUCCUGUGA 
                 523 
                 A-142486 
                 UCACAGGAAAGGAGAAGGU 
                 731 
                 616-634 
               
               
                   
               
               
                 AD-71057 
                 A-142487 
                 CUGUGAGGCACGAAGACAU 
                 524 
                 A-142488 
                 AUGUCUUCGUGCCUCACAG 
                 732 
                 629-647 
               
               
                   
               
               
                 AD-71058 
                 A-142489 
                 GAAGACAUCGAUAAGGGCA 
                 525 
                 A-142490 
                 UGCCCUUAUCGAUGUCUUC 
                 733 
                 640-658 
               
               
                   
               
               
                 AD-71059 
                 A-142491 
                 GAUAAGGGCAUCCUUCUCA 
                 526 
                 A-142492 
                 UGAGAAGGAUGCCCUUAUC 
                 734 
                 649-667 
               
               
                   
               
               
                 AD-71060 
                 A-142493 
                 UUCUCAACUGGACCAAGGA 
                 527 
                 A-142494 
                 UCCUUGGUCCAGUUGAGAA 
                 735 
                 662-680 
               
               
                   
               
               
                 AD-71061 
                 A-142495 
                 ACCAAGGGCUUCAAGGCCU 
                 528 
                 A-142496 
                 AGGCCUUGAAGCCCUUGGU 
                 736 
                 673-691 
               
               
                   
               
               
                 AD-71062 
                 A-142497 
                 CAAGGCCUCAGGAGCAGAA 
                 529 
                 A-142498 
                 UUCUGCUCCUGAGGCCUUG 
                 737 
                 684-702 
               
               
                   
               
               
                 AD-71063 
                 A-142499 
                 AGGAGCAGAAGGGAACAAU 
                 530 
                 A-142500 
                 AUUGUUCCCUUCUGCUCCU 
                 738 
                 693-711 
               
               
                   
               
               
                 AD-71064 
                 A-142501 
                 AACAAUGUCGUGGGGCUUA 
                 531 
                 A-142502 
                 UAAGCCCCACGACAUUGUU 
                 739 
                 706-724 
               
               
                   
               
               
                 AD-71065 
                 A-142503 
                 UGGGGCUUCUGCGAGACGA 
                 532 
                 A-142504 
                 UCGUCUCGCAGAAGCCCCA 
                 740 
                 716-734 
               
               
                   
               
               
                 AD-71066 
                 A-142505 
                 CGAGACGCUAUCAAACGGA 
                 533 
                 A-142506 
                 UCCGUUUGAUAGCGUCUCG 
                 741 
                 727-745 
               
               
                   
               
               
                 AD-71067 
                 A-142507 
                 AAACGGAGAGGGGACUUUA 
                 534 
                 A-142508 
                 UAAAGUCCCCUCUCCGUUU 
                 742 
                 739-757 
               
               
                   
               
               
                 AD-71068 
                 A-142509 
                 GGGACUUUGAAAUGGAUGU 
                 535 
                 A-142510 
                 ACAUCCAUUUCAAAGUCCC 
                 743 
                 749-767 
               
               
                   
               
               
                 AD-71069 
                 A-142513 
                 GCAAUGGUGAAUGACACGA 
                 536 
                 A-142514 
                 UCGUGUCAUUCACCAUUGC 
                 744 
                 772-790 
               
               
                   
               
               
                 AD-71070 
                 A-142515 
                 AAUGACACGGUGGCCACGA 
                 537 
                 A-142516 
                 UCGUGGCCACCGUGUCAUU 
                 745 
                 781-799 
               
               
                   
               
               
                 AD-71071 
                 A-142517 
                 GCCACGAUGAUCUCCUGCU 
                 538 
                 A-142518 
                 AGCAGGAGAUCAUCGUGGC 
                 746 
                 793-811 
               
               
                   
               
               
                 AD-71072 
                 A-142519 
                 UCCUGCUACUACGAAGACA 
                 539 
                 A-142520 
                 UGUCUUCGUAGUAGCAGGA 
                 747 
                 805-823 
               
               
                   
               
               
                 AD-71073 
                 A-142523 
                 AGUGCGAGGUCGGCAUGAU 
                 540 
                 A-142524 
                 AUCAUGCCGACCUCGCACU 
                 748 
                 827-845 
               
               
                   
               
               
                 AD-71074 
                 A-142525 
                 GGCAUGAUCGUGGGCACGA 
                 541 
                 A-142526 
                 UCGUGCCCACGAUCAUGCC 
                 749 
                 838-856 
               
               
                   
               
               
                 AD-71075 
                 A-142527 
                 GUGGGCACGGGCUGCAAUA 
                 542 
                 A-142528 
                 UAUUGCAGCCCGUGCCCAC 
                 750 
                 847-865 
               
               
                   
               
               
                 AD-71076 
                 A-142529 
                 UGCAAUGCCUGCUACAUGA 
                 543 
                 A-142530 
                 UCAUGUAGCAGGCAUUGCA 
                 751 
                 859-877 
               
               
                   
               
               
                 AD-71077 
                 A-142531 
                 UACAUGGAGGAGAUGCAGA 
                 544 
                 A-142532 
                 UCUGCAUCUCCUCCAUGUA 
                 752 
                 871-889 
               
               
                   
               
               
                 AD-71078 
                 A-142533 
                 AGAUGCAGAAUGUGGAGCU 
                 545 
                 A-142534 
                 AGCUCCACAUUCUGCAUCU 
                 753 
                 881-899 
               
               
                   
               
               
                 AD-71079 
                 A-142535 
                 UGUGGAGCUGGUGGAGGGA 
                 546 
                 A-142536 
                 UCCCUCCACCAGCUCCACA 
                 754 
                 891-909 
               
               
                   
               
               
                 AD-71080 
                 A-142537 
                 UGGAGGGGGACGAGGGCCA 
                 547 
                 A-142538 
                 UGGCCCUCGUCCCCCUCCA 
                 755 
                 902-920 
               
               
                   
               
               
                 AD-71081 
                 A-142539 
                 GAGGGCCGCAUGUGCGUCA 
                 548 
                 A-142540 
                 UGACGCACAUGCGGCCCUC 
                 756 
                 913-931 
               
               
                   
               
               
                 AD-71082 
                 A-142541 
                 UGCGUCAAUACCGAGUGGA 
                 549 
                 A-142542 
                 UCCACUCGGUAUUGACGCA 
                 757 
                 925-943 
               
               
                   
               
               
                 AD-71083 
                 A-142543 
                 CGAGUGGGGCGCCUUCGGA 
                 550 
                 A-142544 
                 UCCGAAGGCGCCCCACUCG 
                 758 
                 936-954 
               
               
                   
               
               
                 AD-71084 
                 A-142545 
                 GCCUUCGGGGACUCCGGCA 
                 551 
                 A-142546 
                 UGCCGGAGUCCCCGAAGGC 
                 759 
                 946-964 
               
               
                   
               
               
                 AD-71085 
                 A-142547 
                 UCCGGCGAGCUGGACGAGU 
                 552 
                 A-142548 
                 ACUCGUCCAGCUCGCCGGA 
                 760 
                 958-976 
               
               
                   
               
               
                 AD-71086 
                 A-142549 
                 GACGAGUUCCUGCUGGAGU 
                 553 
                 A-142550 
                 ACUCCAGCAGGAACUCGUC 
                 761 
                 970-988 
               
               
                   
               
               
                 AD-71087 
                 A-142551 
                 UGCUGGAGUAUGACCGCCU 
                 554 
                 A-142552 
                 AGGCGGUCAUACUCCAGCA 
                 762 
                 980-998 
               
               
                   
               
               
                 AD-71088 
                 A-142553 
                 GACCGCCUGGUGGACGAGA 
                 555 
                 A-142554 
                 UCUCGUCCACCAGGCGGUC 
                 763 
                  991-1009 
               
               
                   
               
               
                 AD-71089 
                 A-142555 
                 GGACGAGAGCUCUGCAAAC 
                 556 
                 A-142556 
                 GUUUGCAGAGCUCUCGUCC 
                 764 
                 1002-1020 
               
               
                   
               
               
                 AD-71090 
                 A-142557 
                 UCUGCAAACCCCGGUCAGA 
                 557 
                 A-142558 
                 UCUGACCGGGGUUUGCAGA 
                 765 
                 1012-1030 
               
               
                   
               
               
                 AD-71091 
                 A-142559 
                 GGUCAGCAGCUGUAUGAGA 
                 558 
                 A-142560 
                 UCUCAUACAGCUGCUGACC 
                 766 
                 1024-1042 
               
               
                   
               
               
                 AD-71092 
                 A-142561 
                 UAUGAGAAGCUCAUAGGUA 
                 559 
                 A-142562 
                 UACCUAUGAGCUUCUCAUA 
                 767 
                 1036-1054 
               
               
                   
               
               
                 AD-71093 
                 A-142563 
                 UCAUAGGUGGCAAGUACAU 
                 560 
                 A-142564 
                 AUGUACUUGCCACCUAUGA 
                 768 
                 1046-1064 
               
               
                   
               
               
                 AD-71094 
                 A-142565 
                 AAGUACAUGGGCGAGCUGA 
                 561 
                 A-142566 
                 UCAGCUCGCCCAUGUACUU 
                 769 
                 1057-1075 
               
               
                   
               
               
                 AD-71095 
                 A-142567 
                 GCGAGCUGGUGCGGCUUGU 
                 562 
                 A-142568 
                 ACAAGCCGCACCAGCUCGC 
                 770 
                 1067-1085 
               
               
                   
               
               
                 AD-71096 
                 A-142569 
                 GGCUUGUGCUGCUCAGGCU 
                 563 
                 A-142570 
                 AGCCUGAGCAGCACAAGCC 
                 771 
                 1079-1097 
               
               
                   
               
               
                 AD-71097 
                 A-142571 
                 UCAGGCUCGUGGACGAAAA 
                 564 
                 A-142572 
                 UUUUCGUCCACGAGCCUGA 
                 772 
                 1091-1109 
               
               
                   
               
               
                 AD-69448 
                 A-139832 
                 UGGACGAAAACCUGCUCUU 
                 565 
                 A-139833 
                 AAGAGCAGGUUUUCGUCCA 
                 773 
                 1100-1118 
               
               
                   
               
               
                 AD-71098 
                 A-142573 
                 UGCUCUUCCACGGGGAGGA 
                 566 
                 A-142574 
                 UCCUCCCCGUGGAAGAGCA 
                 774 
                 1112-1130 
               
               
                   
               
               
                 AD-71099 
                 A-142575 
                 GGGAGGCCUCCGAGCAGCU 
                 567 
                 A-142576 
                 AGCUGCUCGGAGGCCUCCC 
                 775 
                 1124-1142 
               
               
                   
               
               
                 AD-71100 
                 A-142577 
                 CGAGCAGCUGCGCACACGA 
                 568 
                 A-142578 
                 UCGUGUGCGCAGCUGCUCG 
                 776 
                 1134-1152 
               
               
                   
               
               
                 AD-71101 
                 A-142581 
                 AGCCUUCGAGACGCGCUUA 
                 569 
                 A-142582 
                 UAAGCGCGUCUCGAAGGCU 
                 777 
                 1155-1173 
               
               
                   
               
               
                 AD-71102 
                 A-142583 
                 CGCUUCGUGUCGCAGGUGA 
                 570 
                 A-142584 
                 UCACCUGCGACACGAAGCG 
                 778 
                 1168-1186 
               
               
                   
               
               
                 AD-71103 
                 A-142585 
                 UCGCAGGUGGAGAGCGACA 
                 571 
                 A-142586 
                 UGUCGCUCUCCACCUGCGA 
                 779 
                 1177-1195 
               
               
                   
               
               
                 AD-71104 
                 A-142587 
                 AGCGACACGGGCGACCGCA 
                 572 
                 A-142588 
                 UGCGGUCGCCCGUGUCGCU 
                 780 
                 1189-1207 
               
               
                   
               
               
                 AD-71105 
                 A-142589 
                 CGACCGCAAGCAGAUCUAA 
                 573 
                 A-142590 
                 UUAGAUCUGCUUGCGGUCG 
                 781 
                 1200-1218 
               
               
                   
               
               
                 AD-71106 
                 A-142591 
                 CAGAUCUACAACAUCCUGA 
                 574 
                 A-142592 
                 UCAGGAUGUUGUAGAUCUG 
                 782 
                 1210-1228 
               
               
                   
               
               
                 AD-71107 
                 A-142593 
                 UCCUGAGCACGCUGGGGCU 
                 575 
                 A-142594 
                 AGCCCCAGCGUGCUCAGGA 
                 783 
                 1223-1241 
               
               
                   
               
               
                 AD-71108 
                 A-142595 
                 CUGGGGCUGCGACCCUCGA 
                 576 
                 A-142596 
                 UCGAGGGUCGCAGCCCCAG 
                 784 
                 1234-1252 
               
               
                   
               
               
                 AD-71109 
                 A-142597 
                 CGACCCUCGACCACCGACU 
                 577 
                 A-142598 
                 AGUCGGUGGUCGAGGGUCG 
                 785 
                 1243-1261 
               
               
                   
               
               
                 AD-71110 
                 A-142599 
                 CACCGACUGCGACAUCGUA 
                 578 
                 A-142600 
                 UACGAUGUCGCAGUCGGUG 
                 786 
                 1254-1272 
               
               
                   
               
               
                 AD-71111 
                 A-142601 
                 CAUCGUGCGCCGCGCCUGA 
                 579 
                 A-142602 
                 UCAGGCGCGGCGCACGAUG 
                 787 
                 1266-1284 
               
               
                   
               
               
                 AD-71112 
                 A-142603 
                 CGCGCCUGCGAGAGCGUGU 
                 580 
                 A-142604 
                 ACACGCUCUCGCAGGCGCG 
                 788 
                 1276-1294 
               
               
                   
               
               
                 AD-71113 
                 A-142605 
                 AGCGUGUCUACGCGCGCUA 
                 581 
                 A-142606 
                 UAGCGCGCGUAGACACGCU 
                 789 
                 1288-1306 
               
               
                   
               
               
                 AD-71114 
                 A-142607 
                 CGCGCUGCGCACAUGUGCU 
                 582 
                 A-142608 
                 AGCACAUGUGCGCAGCGCG 
                 790 
                 1300-1318 
               
               
                   
               
               
                 AD-71115 
                 A-142609 
                 ACAUGUGCUCGGCGGGGCU 
                 583 
                 A-142610 
                 AGCCCCGCCGAGCACAUGU 
                 791 
                 1310-1328 
               
               
                   
               
               
                 AD-71116 
                 A-142611 
                 CGGGGCUGGCGGGCGUCAU 
                 584 
                 A-142612 
                 AUGACGCCCGCCAGCCCCG 
                 792 
                 1322-1340 
               
               
                   
               
               
                 AD-71117 
                 A-142613 
                 CGGGCGUCAUCAACCGCAU 
                 585 
                 A-142614 
                 AUGCGGUUGAUGACGCCCG 
                 793 
                 1331-1349 
               
               
                   
               
               
                 AD-71118 
                 A-142615 
                 AACCGCAUGCGCGAGAGCA 
                 586 
                 A-142616 
                 UGCUCUCGCGCAUGCGGUU 
                 794 
                 1342-1360 
               
               
                   
               
               
                 AD-71119 
                 A-142617 
                 AGAGCCGCAGCGAGGACGU 
                 587 
                 A-142618 
                 ACGUCCUCGCUGCGGCUCU 
                 795 
                 1355-1373 
               
               
                   
               
               
                 AD-71120 
                 A-142619 
                 CGAGGACGUAAUGCGCAUA 
                 588 
                 A-142620 
                 UAUGCGCAUUACGUCCUCG 
                 796 
                 1365-1383 
               
               
                   
               
               
                 AD-71121 
                 A-142621 
                 UGCGCAUCACUGUGGGCGU 
                 589 
                 A-142622 
                 ACGCCCACAGUGAUGCGCA 
                 797 
                 1376-1394 
               
               
                   
               
               
                 AD-71122 
                 A-142623 
                 UGGGCGUGGAUGGCUCCGU 
                 590 
                 A-142624 
                 ACGGAGCCAUCCACGCCCA 
                 798 
                 1388-1406 
               
               
                   
               
               
                 AD-71123 
                 A-142625 
                 UGGCUCCGUGUACAAGCUA 
                 591 
                 A-142626 
                 UAGCUUGUACACGGAGCCA 
                 799 
                 1398-1416 
               
               
                   
               
               
                 AD-71124 
                 A-142627 
                 UACAAGCUGCACCCCAGCU 
                 592 
                 A-142628 
                 AGCUGGGGUGCAGCUUGUA 
                 800 
                 1408-1426 
               
               
                   
               
               
                 AD-71125 
                 A-142629 
                 CCAGCUUCAAGGAGCGGUU 
                 593 
                 A-142630 
                 AACCGCUCCUUGAAGCUGG 
                 801 
                 1421-1439 
               
               
                   
               
               
                 AD-71126 
                 A-142631 
                 AGGAGCGGUUCCAUGCCAA 
                 594 
                 A-142632 
                 UUGGCAUGGAACCGCUCCU 
                 802 
                 1430-1448 
               
               
                   
               
               
                 AD-71127 
                 A-142633 
                 AUGCCAGCGUGCGCAGGCU 
                 595 
                 A-142634 
                 AGCCUGCGCACGCUGGCAU 
                 803 
                 1442-1460 
               
               
                   
               
               
                 AD-71128 
                 A-142635 
                 CGCAGGCUGACGCCCAGCU 
                 596 
                 A-142636 
                 AGCUGGGCGUCAGCCUGCG 
                 804 
                 1453-1471 
               
               
                   
               
               
                 AD-71129 
                 A-142637 
                 CCCAGCUGCGAGAUCACCU 
                 597 
                 A-142638 
                 AGGUGAUCUCGCAGCUGGG 
                 805 
                 1465-1483 
               
               
                   
               
               
                 AD-71130 
                 A-142639 
                 GAGAUCACCUUCAUCGAGU 
                 598 
                 A-142640 
                 ACUCGAUGAAGGUGAUCUC 
                 806 
                 1474-1492 
               
               
                   
               
               
                 AD-71131 
                 A-142641 
                 AUCGAGUCGGAGGAGGGCA 
                 599 
                 A-142642 
                 UGCCCUCCUCCGACUCGAU 
                 807 
                 1486-1504 
               
               
                   
               
               
                 AD-71132 
                 A-142643 
                 AGGAGGGCAGUGGCCGGGA 
                 600 
                 A-142644 
                 UCCCGGCCACUGCCCUCCU 
                 808 
                 1496-1514 
               
               
                   
               
               
                 AD-71133 
                 A-142645 
                 CCGGGGCGCGGCCCUGGUA 
                 601 
                 A-142646 
                 UACCAGGGCCGCGCCCCGG 
                 809 
                 1509-1527 
               
               
                   
               
               
                 AD-71134 
                 A-142647 
                 CCCUGGUCUCGGCGGUGGA 
                 602 
                 A-142648 
                 UCCACCGCCGAGACCAGGG 
                 810 
                 1520-1538 
               
               
                   
               
               
                 AD-71135 
                 A-142649 
                 GCGGUGGCCUGUAAGAAGA 
                 603 
                 A-142650 
                 UCUUCUUACAGGCCACCGC 
                 811 
                 1531-1549 
               
               
                   
               
               
                 AD-71136 
                 A-142651 
                 UAAGAAGGCCUGUAUGCUA 
                 604 
                 A-142652 
                 UAGCAUACAGGCCUUCUUA 
                 812 
                 1542-1560 
               
               
                   
               
               
                 AD-71137 
                 A-142653 
                 CUGUAUGCUGGGCCAGUGA 
                 605 
                 A-142654 
                 UCACUGGCCCAGCAUACAG 
                 813 
                 1551-1569 
               
               
                   
               
               
                 AD-71138 
                 A-142655 
                 CAGUGAGAGCAGUGGCCGA 
                 606 
                 A-142656 
                 UCGGCCACUGCUCUCACUG 
                 814 
                 1564-1582 
               
               
                   
               
               
                 AD-71139 
                 A-142657 
                 CAGUGGCCGCAAGCGCAGA 
                 607 
                 A-142658 
                 UCUGCGCUUGCGGCCACUG 
                 815 
                 1573-1591 
               
               
                   
               
               
                 AD-71140 
                 A-142659 
                 AGCGCAGGGAGGAUGCCAA 
                 608 
                 A-142660 
                 UUGGCAUCCUCCCUGCGCU 
                 816 
                 1584-1602 
               
               
                   
               
               
                 AD-71141 
                 A-142661 
                 UGCCACAGCCCCACAGCAA 
                 609 
                 A-142662 
                 UUGCUGUGGGGCUGUGGCA 
                 817 
                 1597-1615 
               
               
                   
               
               
                 AD-71142 
                 A-142663 
                 CACAGCACCCAGGCUCCAU 
                 610 
                 A-142664 
                 AUGGAGCCUGGGUGCUGUG 
                 818 
                 1608-1626 
               
               
                   
               
               
                 AD-71143 
                 A-142665 
                 AGGCUCCAUGGGGAAGUGA 
                 611 
                 A-142666 
                 UCACUUCCCCAUGGAGCCU 
                 819 
                 1618-1636 
               
               
                   
               
               
                 AD-71144 
                 A-142667 
                 GGAAGUGCUCCCCACACGU 
                 612 
                 A-142668 
                 ACGUGUGGGGAGCACUUCC 
                 820 
                 1629-1647 
               
               
                   
               
               
                 AD-71145 
                 A-142669 
                 CCACACGUGCUCGCAGCCU 
                 613 
                 A-142670 
                 AGGCUGCGAGCACGUGUGG 
                 821 
                 1640-1658 
               
               
                   
               
               
                 AD-71146 
                 A-142671 
                 UCGCAGCCUGGCGGGGCAA 
                 614 
                 A-142672 
                 UUGCCCCGCCAGGCUGCGA 
                 822 
                 1650-1668 
               
               
                   
               
               
                 AD-71147 
                 A-142673 
                 CGGGGCAGGAGGCCUGGCA 
                 615 
                 A-142674 
                 UGCCAGGCCUCCUGCCCCG 
                 823 
                 1661-1679 
               
               
                   
               
               
                 AD-71148 
                 A-142675 
                 CCUGGCCUUGUCAGGACCA 
                 616 
                 A-142676 
                 UGGUCCUGACAAGGCCAGG 
                 824 
                 1673-1691 
               
               
                   
               
               
                 AD-71149 
                 A-142677 
                 CAGGACCCAGGCCGCCUGA 
                 617 
                 A-142678 
                 UCAGGCGGCCUGGGUCCUG 
                 825 
                 1684-1702 
               
               
                   
               
               
                 AD-71150 
                 A-142679 
                 CCGCCUGCCAUACCGCUGA 
                 618 
                 A-142680 
                 UCAGCGGUAUGGCAGGCGG 
                 826 
                 1695-1713 
               
               
                   
               
               
                 AD-71151 
                 A-142681 
                 UACCGCUGGGGAACAGAGA 
                 619 
                 A-142682 
                 UCUCUGUUCCCCAGCGGUA 
                 827 
                 1705-1723 
               
               
                   
               
               
                 AD-71152 
                 A-142683 
                 AACAGAGCGGGCCUCUUCA 
                 620 
                 A-142684 
                 UGAAGAGGCCCGCUCUGUU 
                 828 
                 1716-1734 
               
               
                   
               
               
                 AD-71153 
                 A-142685 
                 CUCUUCCCUCAGUUUUUCA 
                 621 
                 A-142686 
                 UGAAAAACUGAGGGAAGAG 
                 829 
                 1728-1746 
               
               
                   
               
               
                 AD-71154 
                 A-142687 
                 UUUUUCGGUGGGACAGCCA 
                 622 
                 A-142688 
                 UGGCUGUCCCACCGAAAAA 
                 830 
                 1740-1758 
               
               
                   
               
               
                 AD-71155 
                 A-142689 
                 GGGACAGCCCCAGGGCCCU 
                 623 
                 A-142690 
                 AGGGCCCUGGGGCUGUCCC 
                 831 
                 1749-1767 
               
               
                   
               
               
                 AD-71156 
                 A-142691 
                 AGGGCCCUAACGGGGGUGA 
                 624 
                 A-142692 
                 UCACCCCCGUUAGGGCCCU 
                 832 
                 1760-1778 
               
               
                   
               
               
                 AD-71157 
                 A-142693 
                 GGGUGCGGCAGGAGCAGGA 
                 625 
                 A-142694 
                 UCCUGCUCCUGCCGCACCC 
                 833 
                 1773-1791 
               
               
                   
               
               
                 AD-71158 
                 A-142695 
                 AGGAGCAGGAACAGAGACU 
                 626 
                 A-142696 
                 AGUCUCUGUUCCUGCUCCU 
                 834 
                 1782-1800 
               
               
                   
               
               
                 AD-71159 
                 A-142697 
                 AGAGACUCUGGAAGCCCCA 
                 627 
                 A-142698 
                 UGGGGCUUCCAGAGUCUCU 
                 835 
                 1794-1812 
               
               
                   
               
               
                 AD-71160 
                 A-142699 
                 AAGCCCCCCACCUUUCUCA 
                 628 
                 A-142700 
                 UGAGAAAGGUGGGGGGCUU 
                 836 
                 1805-1823 
               
               
                   
               
               
                 AD-71161 
                 A-142701 
                 UUUCUCGCUGGAAUCAAUU 
                 629 
                 A-142702 
                 AAUUGAUUCCAGCGAGAAA 
                 837 
                 1817-1835 
               
               
                   
               
               
                 AD-71162 
                 A-142703 
                 AAUCAAUUUCCCAGAAGGA 
                 630 
                 A-142704 
                 UCCUUCUGGGAAAUUGAUU 
                 838 
                 1828-1846 
               
               
                   
               
               
                 AD-71163 
                 A-142705 
                 CCCAGAAGGGAGUUGCUCA 
                 631 
                 A-142706 
                 UGAGCAACUCCCUUCUGGG 
                 839 
                 1837-1855 
               
               
                   
               
               
                 AD-71164 
                 A-142707 
                 UUGCUCACUCAGGACUUUA 
                 632 
                 A-142708 
                 UAAAGUCCUGAGUGAGCAA 
                 840 
                 1849-1867 
               
               
                   
               
               
                 AD-71165 
                 A-142709 
                 AGGACUUUGAUGCAUUUCA 
                 633 
                 A-142710 
                 UGAAAUGCAUCAAAGUCCU 
                 841 
                 1859-1877 
               
               
                   
               
               
                 AD-71166 
                 A-142711 
                 AUUUCCACACUGUCAGAGA 
                 634 
                 A-142712 
                 UCUCUGACAGUGUGGAAAU 
                 842 
                 1872-1890 
               
               
                   
               
               
                 AD-71167 
                 A-142713 
                 UGUCAGAGCUGUUGGCCUA 
                 635 
                 A-142714 
                 UAGGCCAACAGCUCUGACA 
                 843 
                 1882-1900 
               
               
                   
               
               
                 AD-71168 
                 A-142715 
                 UUGGCCUCGCCUGGGCCCA 
                 636 
                 A-142716 
                 UGGGCCCAGGCGAGGCCAA 
                 844 
                 1893-1911 
               
               
                   
               
               
                 AD-71169 
                 A-142717 
                 CUGGGCCCAGGCUCUGGGA 
                 637 
                 A-142718 
                 UCCCAGAGCCUGGGCCCAG 
                 845 
                 1903-1921 
               
               
                   
               
               
                 AD-71170 
                 A-142719 
                 CUCUGGGAAGGGGUGCCCU 
                 638 
                 A-142720 
                 AGGGCACCCCUUCCCAGAG 
                 846 
                 1914-1932 
               
               
                   
               
               
                 AD-71171 
                 A-142721 
                 UGCCCUCUGGAUCCUGCUA 
                 639 
                 A-142722 
                 UAGCAGGAUCCAGAGGGCA 
                 847 
                 1927-1945 
               
               
                   
               
               
                 AD-71172 
                 A-142723 
                 UCCUGCUGUGGCCUCACUU 
                 640 
                 A-142724 
                 AAGUGAGGCCACAGCAGGA 
                 848 
                 1938-1956 
               
               
                   
               
               
                 AD-71173 
                 A-142725 
                 CCUCACUUCCCUGGGAACU 
                 641 
                 A-142726 
                 AGUUCCCAGGGAAGUGAGG 
                 849 
                 1949-1967 
               
               
                   
               
               
                 AD-71174 
                 A-142727 
                 CUGGGAACUCAUCCUGUGU 
                 642 
                 A-142728 
                 ACACAGGAUGAGUUCCCAG 
                 850 
                 1959-1977 
               
               
                   
               
               
                 AD-71175 
                 A-142729 
                 CCUGUGUGGGGAGGCAGCU 
                 643 
                 A-142730 
                 AGCUGCCUCCCCACACAGG 
                 851 
                 1971-1989 
               
               
                   
               
               
                 AD-71176 
                 A-142731 
                 GGAGGCAGCUCCAACAGCU 
                 644 
                 A-142732 
                 AGCUGUUGGAGCUGCCUCC 
                 852 
                 1980-1998 
               
               
                   
               
               
                 AD-71177 
                 A-142733 
                 CAACAGCUUGACCAGACCU 
                 645 
                 A-142734 
                 AGGUCUGGUCAAGCUGUUG 
                 853 
                 1991-2009 
               
               
                   
               
               
                 AD-71178 
                 A-142735 
                 CCAGACCUAGACCUGGGCA 
                 646 
                 A-142736 
                 UGCCCAGGUCUAGGUCUGG 
                 854 
                 2002-2020 
               
               
                   
               
               
                 AD-71179 
                 A-142737 
                 CUGGGCCAAAAGGGCAGCA 
                 647 
                 A-142738 
                 UGCUGCCCUUUUGGCCCAG 
                 855 
                 2014-2032 
               
               
                   
               
               
                 AD-71180 
                 A-142739 
                 AGGGCAGCCAGGGGCUGCU 
                 648 
                 A-142740 
                 AGCAGCCCCUGGCUGCCCU 
                 856 
                 2024-2042 
               
               
                   
               
               
                 AD-71181 
                 A-142741 
                 GGGCUGCUCAUCACCCAGU 
                 649 
                 A-142742 
                 ACUGGGUGAUGAGCAGCCC 
                 857 
                 2035-2053 
               
               
                   
               
               
                 AD-71182 
                 A-142743 
                 ACCCAGUCCUGGCCAUUUU 
                 650 
                 A-142744 
                 AAAAUGGCCAGGACUGGGU 
                 858 
                 2047-2065 
               
               
                   
               
               
                 AD-71183 
                 A-142745 
                 GCCAUUUUCUUGCCUGAGA 
                 651 
                 A-142746 
                 UCUCAGGCAAGAAAAUGGC 
                 859 
                 2058-2076 
               
               
                   
               
               
                 AD-71184 
                 A-142747 
                 CCUGAGGCUCAAGAGGCCA 
                 652 
                 A-142748 
                 UGGCCUCUUGAGCCUCAGG 
                 860 
                 2070-2088 
               
               
                   
               
               
                 AD-71185 
                 A-142749 
                 AAGAGGCCCAGGGAGCAAU 
                 653 
                 A-142750 
                 AUUGCUCCCUGGGCCUCUU 
                 861 
                 2080-2098 
               
               
                   
               
               
                 AD-71186 
                 A-142751 
                 GGAGCAAUGGGAGGGGGCU 
                 654 
                 A-142752 
                 AGCCCCCUCCCAUUGCUCC 
                 862 
                 2091-2109 
               
               
                   
               
               
                 AD-71187 
                 A-142753 
                 AGGGGGCUCCAUGGAGGAA 
                 655 
                 A-142754 
                 UUCCUCCAUGGAGCCCCCU 
                 863 
                 2102-2120 
               
               
                   
               
               
                 AD-71188 
                 A-142755 
                 GGAGGAGGUGUCCCAAGCU 
                 656 
                 A-142756 
                 AGCUUGGGACACCUCCUCC 
                 864 
                 2114-2132 
               
               
                   
               
               
                 AD-71189 
                 A-142757 
                 UCCCAAGCUUUGAAUACCA 
                 657 
                 A-142758 
                 UGGUAUUCAAAGCUUGGGA 
                 865 
                 2124-2142 
               
               
                   
               
               
                 AD-71190 
                 A-142759 
                 AAUACCCCCAGAGACCUUU 
                 658 
                 A-142760 
                 AAAGGUCUCUGGGGGUAUU 
                 866 
                 2136-2154 
               
               
                   
               
               
                 AD-71191 
                 A-142761 
                 AGAGACCUUUUCUCUCCCA 
                 659 
                 A-142762 
                 UGGGAGAGAAAAGGUCUCU 
                 867 
                 2145-2163 
               
               
                   
               
               
                 AD-71192 
                 A-142763 
                 UCUCCCAUACCAUCACUGA 
                 660 
                 A-142764 
                 UCAGUGAUGGUAUGGGAGA 
                 868 
                 2157-2175 
               
               
                   
               
               
                 AD-71193 
                 A-142765 
                 UCACUGAGUGGCUUGUGAU 
                 661 
                 A-142766 
                 AUCACAAGCCACUCAGUGA 
                 869 
                 2169-2187 
               
               
                   
               
               
                 AD-71194 
                 A-142767 
                 GGCUUGUGAUUCUGGGAUA 
                 662 
                 A-142768 
                 UAUCCCAGAAUCACAAGCC 
                 870 
                 2178-2196 
               
               
                   
               
               
                 AD-71195 
                 A-142769 
                 UGGGAUGGACCCUCGCAGA 
                 663 
                 A-142770 
                 UCUGCGAGGGUCCAUCCCA 
                 871 
                 2190-2208 
               
               
                   
               
               
                 AD-71196 
                 A-142771 
                 UCGCAGCAGGUGCAAGAGA 
                 664 
                 A-142772 
                 UCUCUUGCACCUGCUGCGA 
                 872 
                 2202-2220 
               
               
                   
               
               
                 AD-71197 
                 A-142773 
                 UGCAAGAGACAGAGCCCCA 
                 665 
                 A-142774 
                 UGGGGCUCUGUCUCUUGCA 
                 873 
                 2212-2230 
               
               
                   
               
               
                 AD-71198 
                 A-142775 
                 AGAGCCCCCAAGCCUCUGA 
                 666 
                 A-142776 
                 UCAGAGGCUUGGGGGCUCU 
                 874 
                 2222-2240 
               
               
                   
               
               
                 AD-71199 
                 A-142777 
                 CUCUGCCCCAAGGGGCCCA 
                 667 
                 A-142778 
                 UGGGCCCCUUGGGGCAGAG 
                 875 
                 2235-2253 
               
               
                   
               
               
                 AD-71200 
                 A-142779 
                 AAGGGGCCCACAAAGGGGA 
                 668 
                 A-142780 
                 UCCCCUUUGUGGGCCCCUU 
                 876 
                 2244-2262 
               
               
                   
               
               
                 AD-71201 
                 A-142781 
                 AAAGGGGAGAAGGGCCAGA 
                 669 
                 A-142782 
                 UCUGGCCCUUCUCCCCUUU 
                 877 
                 2255-2273 
               
               
                   
               
               
                 AD-71202 
                 A-142783 
                 GGGCCAGCCCUACAUCUUA 
                 670 
                 A-142784 
                 UAAGAUGUAGGGCUGGCCC 
                 878 
                 2266-2284 
               
               
                   
               
               
                 AD-71203 
                 A-142785 
                 AUCUUCAGCUCCCAUAGCA 
                 671 
                 A-142786 
                 UGCUAUGGGAGCUGAAGAU 
                 879 
                 2279-2297 
               
               
                   
               
               
                 AD-71204 
                 A-142787 
                 UCCCAUAGCGCUGGCUCAA 
                 672 
                 A-142788 
                 UUGAGCCAGCGCUAUGGGA 
                 880 
                 2288-2306 
               
               
                   
               
               
                 AD-71205 
                 A-142789 
                 UGGCUCAGGAAGAAACCCA 
                 673 
                 A-142790 
                 UGGGUUUCUUCCUGAGCCA 
                 881 
                 2299-2317 
               
               
                   
               
               
                 AD-71206 
                 A-142791 
                 AACCCCAAGCAGCAUUCAA 
                 674 
                 A-142792 
                 UUGAAUGCUGCUUGGGGUU 
                 882 
                 2312-2330 
               
               
                   
               
               
                 AD-71207 
                 A-142793 
                 CAGCAUUCAGCACACCCCA 
                 675 
                 A-142794 
                 UGGGGUGUGCUGAAUGCUG 
                 883 
                 2321-2339 
               
               
                   
               
               
                 AD-71208 
                 A-142795 
                 CACCCCAAGGGACAACCCA 
                 676 
                 A-142796 
                 UGGGUUGUCCCUUGGGGUG 
                 884 
                 2333-2351 
               
               
                   
               
               
                 AD-71209 
                 A-142797 
                 ACAACCCCAUCAUAUGACA 
                 677 
                 A-142798 
                 UGUCAUAUGAUGGGGUUGU 
                 885 
                 2344-2362 
               
               
                   
               
               
                 AD-71210 
                 A-142801 
                 ACCCUCUCCAUGCCCAACA 
                 678 
                 A-142802 
                 UGUUGGGCAUGGAGAGGGU 
                 886 
                 2367-2385 
               
               
                   
               
               
                 AD-71211 
                 A-142803 
                 UGCCCAACCUAAGAUUGUA 
                 679 
                 A-142804 
                 UACAAUCUUAGGUUGGGCA 
                 887 
                 2377-2395 
               
               
                   
               
               
                 AD-71212 
                 A-142805 
                 AAGAUUGUGUGGGUUUUUU 
                 680 
                 A-142806 
                 AAAAAACCCACACAAUCUU 
                 888 
                 2387-2405 
               
               
                   
               
               
                 AD-71213 
                 A-142807 
                 UUUUUUAAUUAAAAAUGUU 
                 681 
                 A-142808 
                 AACAUUUUUAAUUAAAAAA 
                 889 
                 2400-2418 
               
               
                   
               
               
                 AD-71214 
                 A-142809 
                 UAAAAAUGUUAAAAGUUUU 
                 682 
                 A-142810 
                 AAAACUUUUAACAUUUUUA 
                 890 
                 2409-2427 
               
               
                   
               
               
                 AD-71215 
                 A-142811 
                 AAAGUUUUAAACAUGAAAA 
                 683 
                 A-142812 
                 UUUUCAUGUUUAAAACUUU 
                 891 
                 2420-2438 
               
               
                   
               
            
           
         
       
     
     
       
         
           
               
             
               
                 TABLE 7 
               
             
            
               
                   
               
               
                 GCK Modified Sequences 
               
            
           
           
               
               
               
               
               
               
               
               
               
            
               
                   
                   
                   
                   
                 Anti- 
                   
                   
                   
                   
               
               
                   
                 Sense 
                 Sense 
                 SEQ 
                 ssense 
                 Antisense 
                 SEQ 
                 mRNA 
                 SEQ 
               
               
                 Duplex 
                 Oligo 
                 Sequence 
                 ID 
                 Oligo 
                 Sequence 
                 ID 
                 target 
                 ID 
               
               
                 Name 
                 Name 
                 (5′-3′) 
                 NO: 
                 Name 
                 (5′-3′) 
                 NO: 
                 sequence 
                 NO: 
               
               
                   
               
            
           
           
               
               
               
               
               
               
               
               
               
            
               
                 AD-71009 
                 A-142377 
                 CUGCCAGCCUCA 
                 892 
                 A-142378 
                 AGCUGCCUGAGG 
                 1100 
                 CUGCCAGCCU 
                 1308 
               
               
                   
                   
                 GGCAGCUdTdT 
                   
                   
                 CUGGCAGdTdT 
                   
                 CAGGCAGCU 
                   
               
               
                   
               
               
                 AD-71010 
                 A-142379 
                 UCAGGCAGCUCU 
                 893 
                 A-142380 
                 UGGAUGGAGAGC 
                 1101 
                 UCAGGCAGCU 
                 1309 
               
               
                   
                   
                 CCAUCCAdTdT 
                   
                   
                 UGCCUGAdTdT 
                   
                 CUCCAUCCA 
                   
               
               
                   
               
               
                 AD-71011 
                 A-142381 
                 CCAUCCAAGCAG 
                 894 
                 A-142382 
                 UCAACGGCUGCU 
                 1102 
                 CCAUCCAAGC 
                 1310 
               
               
                   
                   
                 CCGUUGAdTdT 
                   
                   
                 UGGAUGGdTdT 
                   
                 AGCCGUUGC 
                   
               
               
                   
               
               
                 AD-71012 
                 A-142383 
                 AGCCGUUGCUGC 
                 895 
                 A-142384 
                 UCCUGUGGCAGC 
                 1103 
                 AGCCGUUGCU 
                 1311 
               
               
                   
                   
                 CACAGGAdTdT 
                   
                   
                 AACGGCUdTdT 
                   
                 GCCACAGGC 
                   
               
               
                   
               
               
                 AD-71013 
                 A-142385 
                 ACAGGCGGGCCU 
                 896 
                 A-142386 
                 UAGCGUAAGGCC 
                 1104 
                 ACAGGCGGGC 
                 1312 
               
               
                   
                   
                 UACGCUAdTdT 
                   
                   
                 CGCCUGUdTdT 
                   
                 CUUACGCUC 
                   
               
               
                   
               
               
                 AD-71014 
                 A-142387 
                 UUACGCUCCAAG 
                 897 
                 A-142388 
                 UUGUAGCCUUGG 
                 1105 
                 UUACGCUCCA 
                 1313 
               
               
                   
                   
                 GCUACAAdTdT 
                   
                   
                 AGCGUAAdTdT 
                   
                 AGGCUACAG 
                   
               
               
                   
               
               
                 AD-71015 
                 A-142389 
                 AAGGCUACAGCA 
                 898 
                 A-142390 
                 UAGCACAUGCUG 
                 1106 
                 AAGGCUACAG 
                 1314 
               
               
                   
                   
                 UGUGCUAdTdT 
                   
                   
                 UAGCCUUdTdT 
                   
                 CAUGUGCUA 
                   
               
               
                   
               
               
                 AD-71016 
                 A-142391 
                 UGUGCUAGGCCU 
                 899 
                 A-142392 
                 UCUGCUGAGGCC 
                 1107 
                 UGUGCUAGGC 
                 1315 
               
               
                   
                   
                 CAGCAGAdTdT 
                   
                   
                 UAGCACAdTdT 
                   
                 CUCAGCAGG 
                   
               
               
                   
               
               
                 AD-71017 
                 A-142393 
                 UCAGCAGGCAGG 
                 900 
                 A-142394 
                 AGAUGCUCCUGC 
                 1108 
                 UCAGCAGGCA 
                 1316 
               
               
                   
                   
                 AGCAUCUdTdT 
                   
                   
                 CUGCUGAdTdT 
                   
                 GGAGCAUCU 
                   
               
               
                   
               
               
                 AD-71018 
                 A-142395 
                 AGCAUCUCUGCC 
                 901 
                 A-142396 
                 UUUGGGAGGCAG 
                 1109 
                 AGCAUCUCUG 
                 1317 
               
               
                   
                   
                 UCCCAAAdTdT 
                   
                   
                 AGAUGCUdTdT 
                   
                 CCUCCCAAA 
                   
               
               
                   
               
               
                 AD-71019 
                 A-142397 
                 CCUCCCAAAGCA 
                 902 
                 A-142398 
                 AGGUAGAUGCUU 
                 1110 
                 CCUCCCAAAG 
                 1318 
               
               
                   
                   
                 UCUACCUdTdT 
                   
                   
                 UGGGAGGdTdT 
                   
                 CAUCUACCU 
                   
               
               
                   
               
               
                 AD-71020 
                 A-142401 
                 UAGCCCCUCGGA 
                 903 
                 A-142402 
                 UCAUCUCUCCGA 
                 1111 
                 UAGCCCCUCG 
                 1319 
               
               
                   
                   
                 GAGAUGAdTdT 
                   
                   
                 GGGGCUAdTdT 
                   
                 GAGAGAUGG 
                   
               
               
                   
               
               
                 AD-71021 
                 A-142403 
                 AGAGAUGGCGAU 
                 904 
                 A-142404 
                 UACAUCCAUCGC 
                 1112 
                 AGAGAUGGCG 
                 1320 
               
               
                   
                   
                 GGAUGUAdTdT 
                   
                   
                 CAUCUCUdTdT 
                   
                 AUGGAUGUC 
                   
               
               
                   
               
               
                 AD-71022 
                 A-142405 
                 UGGAUGUCACAA 
                 905 
                 A-142406 
                 UGGCUCCUUGUG 
                 1113 
                 UGGAUGUCAC 
                 1321 
               
               
                   
                   
                 GGAGCCAdTdT 
                   
                   
                 ACAUCCAdTdT 
                   
                 AAGGAGCCA 
                   
               
               
                   
               
               
                 AD-71023 
                 A-142407 
                 AGGAGCCAGGCC 
                 906 
                 A-142408 
                 UUGUCUGGGCCU 
                 1114 
                 AGGAGCCAGG 
                 1322 
               
               
                   
                   
                 CAGACAAdTdT 
                   
                   
                 GGCUCCUdTdT 
                   
                 CCCAGACAG 
                   
               
               
                   
               
               
                 AD-71024 
                 A-142411 
                 ACUCUGGUAGAG 
                 907 
                 A-142412 
                 UGAUCUGCUCUA 
                 1115 
                 ACUCUGGUAG 
                 1323 
               
               
                   
                   
                 CAGAUCAdTdT 
                   
                   
                 CCAGAGUdTdT 
                   
                 AGCAGAUCC 
                   
               
               
                   
               
               
                 AD-71025 
                 A-142413 
                 AGCAGAUCCUGG 
                 908 
                 A-142414 
                 AACUCUGCCAGG 
                 1116 
                 AGCAGAUCCU 
                 1324 
               
               
                   
                   
                 CAGAGUUdTdT 
                   
                   
                 AUCUGCUdTdT 
                   
                 GGCAGAGUU 
                   
               
               
                   
               
               
                 AD-71026 
                 A-142415 
                 CAGAGUUCCAGC 
                 909 
                 A-142416 
                 UCCUGCAGCUGG 
                 1117 
                 CAGAGUUCCA 
                 1325 
               
               
                   
                   
                 UGCAGGAdTdT 
                   
                   
                 AACUCUGdTdT 
                   
                 GCUGCAGGA 
                   
               
               
                   
               
               
                 AD-71027 
                 A-142417 
                 AGCUGCAGGAGG 
                 910 
                 A-142418 
                 AGGUCCUCCUCC 
                 1118 
                 AGCUGCAGGA 
                 1326 
               
               
                   
                   
                 AGGACCUdTdT 
                   
                   
                 UGCAGCUdTdT 
                   
                 GGAGGACCU 
                   
               
               
                   
               
               
                 AD-71028 
                 A-142419 
                 AGGACCUGAAGA 
                 911 
                 A-142420 
                 AUCACCUUCUUC 
                 1119 
                 AGGACCUGAA 
                 1327 
               
               
                   
                   
                 AGGUGAUdTdT 
                   
                   
                 AGGUCCUdTdT 
                   
                 GAAGGUGAU 
                   
               
               
                   
               
               
                 AD-71029 
                 A-142421 
                 AAGGUGAUGAGA 
                 912 
                 A-142422 
                 UCAUCCGUCUCA 
                 1120 
                 AAGGUGAUGA 
                 1328 
               
               
                   
                   
                 CGGAUGAdTdT 
                   
                   
                 UCACCUUdTdT 
                   
                 GACGGAUGC 
                   
               
               
                   
               
               
                 AD-71030 
                 A-142423 
                 CGGAUGCAGAAG 
                 913 
                 A-142424 
                 UCAUCUCCUUCU 
                 1121 
                 CGGAUGCAGA 
                 1329 
               
               
                   
                   
                 GAGAUGAdTdT 
                   
                   
                 GCAUCCGdTdT 
                   
                 AGGAGAUGG 
                   
               
               
                   
               
               
                 AD-71031 
                 A-142425 
                 AAGGAGAUGGAC 
                 914 
                 A-142426 
                 UGCCGCGGUCCA 
                 1122 
                 AAGGAGAUGG 
                 1330 
               
               
                   
                   
                 CGCGGCAdTdT 
                   
                   
                 UCUCCUUdTdT 
                   
                 ACCGCGGCC 
                   
               
               
                   
               
               
                 AD-71032 
                 A-142427 
                 CGCGGCCUGAGG 
                 915 
                 A-142428 
                 UCUCCAGCCUCA 
                 1123 
                 CGCGGCCUGA 
                 1331 
               
               
                   
                   
                 CUGGAGAdTdT 
                   
                   
                 GGCCGCGdTdT 
                   
                 GGCUGGAGA 
                   
               
               
                   
               
               
                 AD-71033 
                 A-142429 
                 CUGGAGACCCAU 
                 916 
                 A-142430 
                 UCUCUUCAUGGG 
                 1124 
                 CUGGAGACCC 
                 1332 
               
               
                   
                   
                 GAAGAGAdTdT 
                   
                   
                 UCUCCAGdTdT 
                   
                 AUGAAGAGG 
                   
               
               
                   
               
               
                 AD-71034 
                 A-142431 
                 CAUGAAGAGGCC 
                 917 
                 A-142432 
                 UCACACUGGCCU 
                 1125 
                 CAUGAAGAGG 
                 1333 
               
               
                   
                   
                 AGUGUGAdTdT 
                   
                   
                 CUUCAUGdTdT 
                   
                 CCAGUGUGA 
                   
               
               
                   
               
               
                 AD-71035 
                 A-142433 
                 CAGUGUGAAGAU 
                 918 
                 A-142434 
                 UGGCAGCAUCUU 
                 1126 
                 CAGUGUGAAG 
                 1334 
               
               
                   
                   
                 GCUGCCAdTdT 
                   
                   
                 CACACUGdTdT 
                   
                 AUGCUGCCC 
                   
               
               
                   
               
               
                 AD-71036 
                 A-142435 
                 UGCUGCCCACCU 
                 919 
                 A-142436 
                 UGCACGUAGGUG 
                 1127 
                 UGCUGCCCAC 
                 1335 
               
               
                   
                   
                 ACGUGCAdTdT 
                   
                   
                 GGCAGCAdTdT 
                   
                 CUACGUGCG 
                   
               
               
                   
               
               
                 AD-71037 
                 A-142437 
                 UACGUGCGCUCC 
                 920 
                 A-142438 
                 UUGGGGUGGAGC 
                 1128 
                 UACGUGCGCU 
                 1336 
               
               
                   
                   
                 ACCCCAAdTdT 
                   
                   
                 GCACGUAdTdT 
                   
                 CCACCCCAG 
                   
               
               
                   
               
               
                 AD-71038 
                 A-142439 
                 ACCCCAGAAGGC 
                 921 
                 A-142440 
                 UUUCUGAGCCUU 
                 1129 
                 ACCCCAGAAG 
                 1337 
               
               
                   
                   
                 UCAGAAAdTdT 
                   
                   
                 CUGGGGUdTdT 
                   
                 GCUCAGAAG 
                   
               
               
                   
               
               
                 AD-71039 
                 A-142441 
                 UCAGAAGUCGGG 
                 922 
                 A-142442 
                 UGAAGUCCCCGA 
                 1130 
                 UCAGAAGUCG 
                 1338 
               
               
                   
                   
                 GACUUCAdTdT 
                   
                   
                 CUUCUGAdTdT 
                   
                 GGGACUUCC 
                   
               
               
                   
               
               
                 AD-71040 
                 A-142443 
                 GGGGACUUCCUC 
                 923 
                 A-142444 
                 UCAGGGAGAGGA 
                 1131 
                 GGGGACUUCC 
                 1339 
               
               
                   
                   
                 UCCCUGAdTdT 
                   
                   
                 AGUCCCCdTdT 
                   
                 UCUCCCUGG 
                   
               
               
                   
               
               
                 AD-71041 
                 A-142445 
                 UCCCUGGACCUG 
                 924 
                 A-142446 
                 UGCCACCCAGGU 
                 1132 
                 UCCCUGGACC 
                 1340 
               
               
                   
                   
                 GGUGGCAdTdT 
                   
                   
                 CCAGGGAdTdT 
                   
                 UGGGUGGCA 
                   
               
               
                   
               
               
                 AD-71042 
                 A-142447 
                 UGGGUGGCACUA 
                 925 
                 A-142448 
                 UUGAAGUUAGUG 
                 1133 
                 UGGGUGGCAC 
                 1341 
               
               
                   
                   
                 ACUUCAAdTdT 
                   
                   
                 CCACCCAdTdT 
                   
                 UAACUUCAG 
                   
               
               
                   
               
               
                 AD-71043 
                 A-142449 
                 ACUUCAGGGUGA 
                 926 
                 A-142450 
                 ACCAGCAUCACC 
                 1134 
                 ACUUCAGGGU 
                 1342 
               
               
                   
                   
                 UGCUGGUdTdT 
                   
                   
                 CUGAAGUdTdT 
                   
                 GAUGCUGGU 
                   
               
               
                   
               
               
                 AD-71044 
                 A-142453 
                 AGGUGGGAGAAG 
                 927 
                 A-142454 
                 UCCUCACCUUCU 
                 1135 
                 AGGUGGGAGA 
                 1343 
               
               
                   
                   
                 GUGAGGAdTdT 
                   
                   
                 CCCACCUdTdT 
                   
                 AGGUGAGGA 
                   
               
               
                   
               
               
                 AD-71045 
                 A-142457 
                 CAGUGGAGCGUG 
                 928 
                 A-142458 
                 UGGUCUUCACGC 
                 1136 
                 CAGUGGAGCG 
                 1344 
               
               
                   
                   
                 AAGACCAdTdT 
                   
                   
                 UCCACUGdTdT 
                   
                 UGAAGACCA 
                   
               
               
                   
               
               
                 AD-71046 
                 A-142461 
                 CCAGAUGUACUC 
                 929 
                 A-142462 
                 UGGGAUGGAGUA 
                 1137 
                 CCAGAUGUAC 
                 1345 
               
               
                   
                   
                 CAUCCCAdTdT 
                   
                   
                 CAUCUGGdTdT 
                   
                 UCCAUCCCC 
                   
               
               
                   
               
               
                 AD-71047 
                 A-142467 
                 ACCGGCACUGCU 
                 930 
                 A-142468 
                 UCAUCUCAGCAG 
                 1138 
                 ACCGGCACUG 
                 1346 
               
               
                   
                   
                 GAGAUGAdTdT 
                   
                   
                 UGCCGGUdTdT 
                   
                 CUGAGAUGC 
                   
               
               
                   
               
               
                 AD-71048 
                 A-142469 
                 AGAUGCUCUUCG 
                 931 
                 A-142470 
                 AUGUAGUCGAAG 
                 1139 
                 AGAUGCUCUU 
                 1347 
               
               
                   
                   
                 ACUACAUdTdT 
                   
                   
                 AGCAUCUdTdT 
                   
                 CGACUACAU 
                   
               
               
                   
               
               
                 AD-71049 
                 A-142471 
                 UCGACUACAUCU 
                 932 
                 A-142472 
                 UACUCAGAGAUG 
                 1140 
                 UCGACUACAU 
                 1348 
               
               
                   
                   
                 CUGAGUAdTdT 
                   
                   
                 UAGUCGAdTdT 
                   
                 CUCUGAGUG 
                   
               
               
                   
               
               
                 AD-71050 
                 A-142473 
                 UCUGAGUGCAUC 
                 933 
                 A-142474 
                 AGUCGGAGAUGC 
                 1141 
                 UCUGAGUGCA 
                 1349 
               
               
                   
                   
                 UCCGACUdTdT 
                   
                   
                 ACUCAGAdTdT 
                   
                 UCUCCGACU 
                   
               
               
                   
               
               
                 AD-71051 
                 A-142475 
                 UCCGACUUCCUG 
                 934 
                 A-142476 
                 UCUUGUCCAGGA 
                 1142 
                 UCCGACUUCC 
                 1350 
               
               
                   
                   
                 GACAAGAdTdT 
                   
                   
                 AGUCGGAdTdT 
                   
                 UGGACAAGC 
                   
               
               
                   
               
               
                 AD-71052 
                 A-142477 
                 GACAAGCAUCAG 
                 935 
                 A-142478 
                 GUUUCAUCUGAU 
                 1143 
                 GACAAGCAUC 
                 1351 
               
               
                   
                   
                 AUGAAACdTdT 
                   
                   
                 GCUUGUCdTdT 
                   
                 AGAUGAAAC 
                   
               
               
                   
               
               
                 AD-71053 
                 A-142479 
                 AGAUGAAACACA 
                 936 
                 A-142480 
                 AGCUUCUUGUGU 
                 1144 
                 AGAUGAAACA 
                 1352 
               
               
                   
                   
                 AGAAGCUdTdT 
                   
                   
                 UUCAUCUdTdT 
                   
                 CAAGAAGCU 
                   
               
               
                   
               
               
                 AD-71054 
                 A-142481 
                 AGAAGCUGCCCC 
                 937 
                 A-142482 
                 AAGCCCAGGGGC 
                 1145 
                 AGAAGCUGCC 
                 1353 
               
               
                   
                   
                 UGGGCUUdTdT 
                   
                   
                 AGCUUCUdTdT 
                   
                 CCUGGGCUU 
                   
               
               
                   
               
               
                 AD-71055 
                 A-142483 
                 CCUGGGCUUCAC 
                 938 
                 A-142484 
                 UGAGAAGGUGAA 
                 1146 
                 CCUGGGCUUC 
                 1354 
               
               
                   
                   
                 CUUCUCAdTdT 
                   
                   
                 GCCCAGGdTdT 
                   
                 ACCUUCUCC 
                   
               
               
                   
               
               
                 AD-71056 
                 A-142485 
                 ACCUUCUCCUUU 
                 939 
                 A-142486 
                 UCACAGGAAAGG 
                 1147 
                 ACCUUCUCCU 
                 1355 
               
               
                   
                   
                 CCUGUGAdTdT 
                   
                   
                 AGAAGGUdTdT 
                   
                 UUCCUGUGA 
                   
               
               
                   
               
               
                 AD-71057 
                 A-142487 
                 CUGUGAGGCACG 
                 940 
                 A-142488 
                 AUGUCUUCGUGC 
                 1148 
                 CUGUGAGGCA 
                 1356 
               
               
                   
                   
                 AAGACAUdTdT 
                   
                   
                 CUCACAGdTdT 
                   
                 CGAAGACAU 
                   
               
               
                   
               
               
                 AD-71058 
                 A-142489 
                 GAAGACAUCGAU 
                 941 
                 A-142490 
                 UGCCCUUAUCGA 
                 1149 
                 GAAGACAUCG 
                 1357 
               
               
                   
                   
                 AAGGGCAdTdT 
                   
                   
                 UGUCUUCdTdT 
                   
                 AUAAGGGCA 
                   
               
               
                   
               
               
                 AD-71059 
                 A-142491 
                 GAUAAGGGCAUC 
                 942 
                 A-142492 
                 UGAGAAGGAUGC 
                 1150 
                 GAUAAGGGCA 
                 1358 
               
               
                   
                   
                 CUUCUCAdTdT 
                   
                   
                 CCUUAUCdTdT 
                   
                 UCCUUCUCA 
                   
               
               
                   
               
               
                 AD-71060 
                 A-142493 
                 UUCUCAACUGGA 
                 943 
                 A-142494 
                 UCCUUGGUCCAG 
                 1151 
                 UUCUCAACUG 
                 1359 
               
               
                   
                   
                 CCAAGGAdTdT 
                   
                   
                 UUGAGAAdTdT 
                   
                 GACCAAGGG 
                   
               
               
                   
               
               
                 AD-71061 
                 A-142495 
                 ACCAAGGGCUUC 
                 944 
                 A-142496 
                 AGGCCUUGAAGC 
                 1152 
                 ACCAAGGGCU 
                 1360 
               
               
                   
                   
                 AAGGCCUdTdT 
                   
                   
                 CCUUGGUdTdT 
                   
                 UCAAGGCCU 
                   
               
               
                   
               
               
                 AD-71062 
                 A-142497 
                 CAAGGCCUCAGG 
                 945 
                 A-142498 
                 UUCUGCUCCUGA 
                 1153 
                 CAAGGCCUCA 
                 1361 
               
               
                   
                   
                 AGCAGAAdTdT 
                   
                   
                 GGCCUUGdTdT 
                   
                 GGAGCAGAA 
                   
               
               
                   
               
               
                 AD-71063 
                 A-142499 
                 AGGAGCAGAAGG 
                 946 
                 A-142500 
                 AUUGUUCCCUUC 
                 1154 
                 AGGAGCAGAA 
                 1362 
               
               
                   
                   
                 GAACAAUdTdT 
                   
                   
                 UGCUCCUdTdT 
                   
                 GGGAACAAU 
                   
               
               
                   
               
               
                 AD-71064 
                 A-142501 
                 AACAAUGUCGUG 
                 947 
                 A-142502 
                 UAAGCCCCACGA 
                 1155 
                 AACAAUGUCG 
                 1363 
               
               
                   
                   
                 GGGCUUAdTdT 
                   
                   
                 CAUUGUUdTdT 
                   
                 UGGGGCUUC 
                   
               
               
                   
               
               
                 AD-71065 
                 A-142503 
                 UGGGGCUUCUGC 
                 948 
                 A-142504 
                 UCGUCUCGCAGA 
                 1156 
                 UGGGGCUUCU 
                 1364 
               
               
                   
                   
                 GAGACGAdTdT 
                   
                   
                 AGCCCCAdTdT 
                   
                 GCGAGACGC 
                   
               
               
                   
               
               
                 AD-71066 
                 A-142505 
                 CGAGACGCUAUC 
                 949 
                 A-142506 
                 UCCGUUUGAUAG 
                 1157 
                 CGAGACGCUA 
                 1365 
               
               
                   
                   
                 AAACGGAdTdT 
                   
                   
                 CGUCUCGdTdT 
                   
                 UCAAACGGA 
                   
               
               
                   
               
               
                 AD-71067 
                 A-142507 
                 AAACGGAGAGGG 
                 950 
                 A-142508 
                 UAAAGUCCCCUC 
                 1158 
                 AAACGGAGAG 
                 1366 
               
               
                   
                   
                 GACUUUAdTdT 
                   
                   
                 UCCGUUUdTdT 
                   
                 GGGACUUUG 
                   
               
               
                   
               
               
                 AD-71068 
                 A-142509 
                 GGGACUUUGAAA 
                 951 
                 A-142510 
                 ACAUCCAUUUCA 
                 1159 
                 GGGACUUUGA 
                 1367 
               
               
                   
                   
                 UGGAUGUdTdT 
                   
                   
                 AAGUCCCdTdT 
                   
                 AAUGGAUGU 
                   
               
               
                   
               
               
                 AD-71069 
                 A-142513 
                 GCAAUGGUGAAU 
                 952 
                 A-142514 
                 UCGUGUCAUUCA 
                 1160 
                 GCAAUGGUGA 
                 1368 
               
               
                   
                   
                 GACACGAdTdT 
                   
                   
                 CCAUUGCdTdT 
                   
                 AUGACACGG 
                   
               
               
                   
               
               
                 AD-71070 
                 A-142515 
                 AAUGACACGGUG 
                 953 
                 A-142516 
                 UCGUGGCCACCG 
                 1161 
                 AAUGACACGG 
                 1369 
               
               
                   
                   
                 GCCACGAdTdT 
                   
                   
                 UGUCAUUdTdT 
                   
                 UGGCCACGA 
                   
               
               
                   
               
               
                 AD-71071 
                 A-142517 
                 GCCACGAUGAUC 
                 954 
                 A-142518 
                 AGCAGGAGAUCA 
                 1162 
                 GCCACGAUGA 
                 1370 
               
               
                   
                   
                 UCCUGCUdTdT 
                   
                   
                 UCGUGGCdTdT 
                   
                 UCUCCUGCU 
                   
               
               
                   
               
               
                 AD-71072 
                 A-142519 
                 UCCUGCUACUAC 
                 955 
                 A-142520 
                 UGUCUUCGUAGU 
                 1163 
                 UCCUGCUACU 
                 1371 
               
               
                   
                   
                 GAAGACAdTdT 
                   
                   
                 AGCAGGAdTdT 
                   
                 ACGAAGACC 
                   
               
               
                   
               
               
                 AD-71073 
                 A-142523 
                 AGUGCGAGGUCG 
                 956 
                 A-142524 
                 AUCAUGCCGACC 
                 1164 
                 AGUGCGAGGU 
                 1372 
               
               
                   
                   
                 GCAUGAUdTdT 
                   
                   
                 UCGCACUdTdT 
                   
                 CGGCAUGAU 
                   
               
               
                   
               
               
                 AD-71074 
                 A-142525 
                 GGCAUGAUCGUG 
                 957 
                 A-142526 
                 UCGUGCCCACGA 
                 1165 
                 GGCAUGAUCG 
                 1373 
               
               
                   
                   
                 GGCACGAdTdT 
                   
                   
                 UCAUGCCdTdT 
                   
                 UGGGCACGG 
                   
               
               
                   
               
               
                 AD-71075 
                 A-142527 
                 GUGGGCACGGGC 
                 958 
                 A-142528 
                 UAUUGCAGCCCG 
                 1166 
                 GUGGGCACGG 
                 1374 
               
               
                   
                   
                 UGCAAUAdTdT 
                   
                   
                 UGCCCACdTdT 
                   
                 GCUGCAAUG 
                   
               
               
                   
               
               
                 AD-71076 
                 A-142529 
                 UGCAAUGCCUGC 
                 959 
                 A-142530 
                 UCAUGUAGCAGG 
                 1167 
                 UGCAAUGCCU 
                 1375 
               
               
                   
                   
                 UACAUGAdTdT 
                   
                   
                 CAUUGCAdTdT 
                   
                 GCUACAUGG 
                   
               
               
                   
               
               
                 AD-71077 
                 A-142531 
                 UACAUGGAGGAG 
                 960 
                 A-142532 
                 UCUGCAUCUCCU 
                 1168 
                 UACAUGGAGG 
                 1376 
               
               
                   
                   
                 AUGCAGAdTdT 
                   
                   
                 CCAUGUAdTdT 
                   
                 AGAUGCAGA 
                   
               
               
                   
               
               
                 AD-71078 
                 A-142533 
                 AGAUGCAGAAUG 
                 961 
                 A-142534 
                 AGCUCCACAUUC 
                 1169 
                 AGAUGCAGAA 
                 1377 
               
               
                   
                   
                 UGGAGCUdTdT 
                   
                   
                 UGCAUCUdTdT 
                   
                 UGUGGAGCU 
                   
               
               
                   
               
               
                 AD-71079 
                 A-142535 
                 UGUGGAGCUGGU 
                 962 
                 A-142536 
                 UCCCUCCACCAG 
                 1170 
                 UGUGGAGCUG 
                 1378 
               
               
                   
                   
                 GGAGGGAdTdT 
                   
                   
                 CUCCACAdTdT 
                   
                 GUGGAGGGG 
                   
               
               
                   
               
               
                 AD-71080 
                 A-142537 
                 UGGAGGGGGACG 
                 963 
                 A-142538 
                 UGGCCCUCGUCC 
                 1171 
                 UGGAGGGGGA 
                 1379 
               
               
                   
                   
                 AGGGCCAdTdT 
                   
                   
                 CCCUCCAdTdT 
                   
                 CGAGGGCCG 
                   
               
               
                   
               
               
                 AD-71081 
                 A-142539 
                 GAGGGCCGCAUG 
                 964 
                 A-142540 
                 UGACGCACAUGC 
                 1172 
                 GAGGGCCGCA 
                 1380 
               
               
                   
                   
                 UGCGUCAdTdT 
                   
                   
                 GGCCCUCdTdT 
                   
                 UGUGCGUCA 
                   
               
               
                   
               
               
                 AD-71082 
                 A-142541 
                 UGCGUCAAUACC 
                 965 
                 A-142542 
                 UCCACUCGGUAU 
                 1173 
                 UGCGUCAAUA 
                 1381 
               
               
                   
                   
                 GAGUGGAdTdT 
                   
                   
                 UGACGCAdTdT 
                   
                 CCGAGUGGG 
                   
               
               
                   
               
               
                 AD-71083 
                 A-142543 
                 CGAGUGGGGCGC 
                 966 
                 A-142544 
                 UCCGAAGGCGCC 
                 1174 
                 CGAGUGGGGC 
                 1382 
               
               
                   
                   
                 CUUCGGAdTdT 
                   
                   
                 CCACUCGdTdT 
                   
                 GCCUUCGGG 
                   
               
               
                   
               
               
                 AD-71084 
                 A-142545 
                 GCCUUCGGGGAC 
                 967 
                 A-142546 
                 UGCCGGAGUCCC 
                 1175 
                 GCCUUCGGGG 
                 1383 
               
               
                   
                   
                 UCCGGCAdTdT 
                   
                   
                 CGAAGGCdTdT 
                   
                 ACUCCGGCG 
                   
               
               
                   
               
               
                 AD-71085 
                 A-142547 
                 UCCGGCGAGCUG 
                 968 
                 A-142548 
                 ACUCGUCCAGCU 
                 1176 
                 UCCGGCGAGC 
                 1384 
               
               
                   
                   
                 GACGAGUdTdT 
                   
                   
                 CGCCGGAdTdT 
                   
                 UGGACGAGU 
                   
               
               
                   
               
               
                 AD-71086 
                 A-142549 
                 GACGAGUUCCUG 
                 969 
                 A-142550 
                 ACUCCAGCAGGA 
                 1177 
                 GACGAGUUCC 
                 1385 
               
               
                   
                   
                 CUGGAGUdTdT 
                   
                   
                 ACUCGUCdTdT 
                   
                 UGCUGGAGU 
                   
               
               
                   
               
               
                 AD-71087 
                 A-142551 
                 UGCUGGAGUAUG 
                 970 
                 A-142552 
                 AGGCGGUCAUAC 
                 1178 
                 UGCUGGAGUA 
                 1386 
               
               
                   
                   
                 ACCGCCUdTdT 
                   
                   
                 UCCAGCAdTdT 
                   
                 UGACCGCCU 
                   
               
               
                   
               
               
                 AD-71088 
                 A-142553 
                 GACCGCCUGGUG 
                 971 
                 A-142554 
                 UCUCGUCCACCA 
                 1179 
                 GACCGCCUGG 
                 1387 
               
               
                   
                   
                 GACGAGAdTdT 
                   
                   
                 GGCGGUCdTdT 
                   
                 UGGACGAGA 
                   
               
               
                   
               
               
                 AD-71089 
                 A-142555 
                 GGACGAGAGCUC 
                 972 
                 A-142556 
                 GUUUGCAGAGCU 
                 1180 
                 GGACGAGAGC 
                 1388 
               
               
                   
                   
                 UGCAAACdTdT 
                   
                   
                 CUCGUCCdTdT 
                   
                 UCUGCAAAC 
                   
               
               
                   
               
               
                 AD-71090 
                 A-142557 
                 UCUGCAAACCCC 
                 973 
                 A-142558 
                 UCUGACCGGGGU 
                 1181 
                 UCUGCAAACC 
                 1389 
               
               
                   
                   
                 GGUCAGAdTdT 
                   
                   
                 UUGCAGAdTdT 
                   
                 CCGGUCAGC 
                   
               
               
                   
               
               
                 AD-71091 
                 A-142559 
                 GGUCAGCAGCUG 
                 974 
                 A-142560 
                 UCUCAUACAGCU 
                 1182 
                 GGUCAGCAGC 
                 1390 
               
               
                   
                   
                 UAUGAGAdTdT 
                   
                   
                 GCUGACCdTdT 
                   
                 UGUAUGAGA 
                   
               
               
                   
               
               
                 AD-71092 
                 A-142561 
                 UAUGAGAAGCUC 
                 975 
                 A-142562 
                 UACCUAUGAGCU 
                 1183 
                 UAUGAGAAGC 
                 1391 
               
               
                   
                   
                 AUAGGUAdTdT 
                   
                   
                 UCUCAUAdTdT 
                   
                 UCAUAGGUG 
                   
               
               
                   
               
               
                 AD-71093 
                 A-142563 
                 UCAUAGGUGGCA 
                 976 
                 A-142564 
                 AUGUACUUGCCA 
                 1184 
                 UCAUAGGUGG 
                 1392 
               
               
                   
                   
                 AGUACAUdTdT 
                   
                   
                 CCUAUGAdTdT 
                   
                 CAAGUACAU 
                   
               
               
                   
               
               
                 AD-71094 
                 A-142565 
                 AAGUACAUGGGC 
                 977 
                 A-142566 
                 UCAGCUCGCCCA 
                 1185 
                 AAGUACAUGG 
                 1393 
               
               
                   
                   
                 GAGCUGAdTdT 
                   
                   
                 UGUACUUdTdT 
                   
                 GCGAGCUGG 
                   
               
               
                   
               
               
                 AD-71095 
                 A-142567 
                 GCGAGCUGGUGC 
                 978 
                 A-142568 
                 ACAAGCCGCACC 
                 1186 
                 GCGAGCUGGU 
                 1394 
               
               
                   
                   
                 GGCUUGUdTdT 
                   
                   
                 AGCUCGCdTdT 
                   
                 GCGGCUUGU 
                   
               
               
                   
               
               
                 AD-71096 
                 A-142569 
                 GGCUUGUGCUGC 
                 979 
                 A-142570 
                 AGCCUGAGCAGC 
                 1187 
                 GGCUUGUGCU 
                 1395 
               
               
                   
                   
                 UCAGGCUdTdT 
                   
                   
                 ACAAGCCdTdT 
                   
                 GCUCAGGCU 
                   
               
               
                   
               
               
                 AD-71097 
                 A-142571 
                 UCAGGCUCGUGG 
                 980 
                 A-142572 
                 UUUUCGUCCACG 
                 1188 
                 UCAGGCUCGU 
                 1396 
               
               
                   
                   
                 ACGAAAAdTdT 
                   
                   
                 AGCCUGAdTdT 
                   
                 GGACGAAAA 
                   
               
               
                   
               
               
                 AD-69448 
                 A-139832 
                 UGGACGAAAACC 
                 981 
                 A-139833 
                 AAGAGCAGGUUU 
                 1189 
                 UGGACGAAAA 
                 1397 
               
               
                   
                   
                 UGCUCUUdTdT 
                   
                   
                 UCGUCCAdTdT 
                   
                 CCUGCUCUU 
                   
               
               
                   
               
               
                 AD-71098 
                 A-142573 
                 UGCUCUUCCACG 
                 982 
                 A-142574 
                 UCCUCCCCGUGG 
                 1190 
                 UGCUCUUCCA 
                 1398 
               
               
                   
                   
                 GGGAGGAdTdT 
                   
                   
                 AAGAGCAdTdT 
                   
                 CGGGGAGGC 
                   
               
               
                   
               
               
                 AD-71099 
                 A-142575 
                 GGGAGGCCUCCG 
                 983 
                 A-142576 
                 AGCUGCUCGGAG 
                 1191 
                 GGGAGGCCUC 
                 1399 
               
               
                   
                   
                 AGCAGCUdTdT 
                   
                   
                 GCCUCCCdTdT 
                   
                 CGAGCAGCU 
                   
               
               
                   
               
               
                 AD-71100 
                 A-142577 
                 CGAGCAGCUGCG 
                 984 
                 A-142578 
                 UCGUGUGCGCAG 
                 1192 
                 CGAGCAGCUG 
                 1400 
               
               
                   
                   
                 CACACGAdTdT 
                   
                   
                 CUGCUCGdTdT 
                   
                 CGCACACGC 
                   
               
               
                   
               
               
                 AD-71101 
                 A-142581 
                 AGCCUUCGAGAC 
                 985 
                 A-142582 
                 UAAGCGCGUCUC 
                 1193 
                 AGCCUUCGAG 
                 1401 
               
               
                   
                   
                 GCGCUUAdTdT 
                   
                   
                 GAAGGCUdTdT 
                   
                 ACGCGCUUC 
                   
               
               
                   
               
               
                 AD-71102 
                 A-142583 
                 CGCUUCGUGUCG 
                 986 
                 A-142584 
                 UCACCUGCGACA 
                 1194 
                 CGCUUCGUGU 
                 1402 
               
               
                   
                   
                 CAGGUGAdTdT 
                   
                   
                 CGAAGCGdTdT 
                   
                 CGCAGGUGG 
                   
               
               
                   
               
               
                 AD-71103 
                 A-142585 
                 UCGCAGGUGGAG 
                 987 
                 A-142586 
                 UGUCGCUCUCCA 
                 1195 
                 UCGCAGGUGG 
                 1403 
               
               
                   
                   
                 AGCGACAdTdT 
                   
                   
                 CCUGCGAdTdT 
                   
                 AGAGCGACA 
                   
               
               
                   
               
               
                 AD-71104 
                 A-142587 
                 AGCGACACGGGC 
                 988 
                 A-142588 
                 UGCGGUCGCCCG 
                 1196 
                 AGCGACACGG 
                 1404 
               
               
                   
                   
                 GACCGCAdTdT 
                   
                   
                 UGUCGCUdTdT 
                   
                 GCGACCGCA 
                   
               
               
                   
               
               
                 AD-71105 
                 A-142589 
                 CGACCGCAAGCA 
                 989 
                 A-142590 
                 UUAGAUCUGCUU 
                 1197 
                 CGACCGCAAG 
                 1405 
               
               
                   
                   
                 GAUCUAAdTdT 
                   
                   
                 GCGGUCGdTdT 
                   
                 CAGAUCUAC 
                   
               
               
                   
               
               
                 AD-71106 
                 A-142591 
                 CAGAUCUACAAC 
                 990 
                 A-142592 
                 UCAGGAUGUUGU 
                 1198 
                 CAGAUCUACA 
                 1406 
               
               
                   
                   
                 AUCCUGAdTdT 
                   
                   
                 AGAUCUGdTdT 
                   
                 ACAUCCUGA 
                   
               
               
                   
               
               
                 AD-71107 
                 A-142593 
                 UCCUGAGCACGC 
                 991 
                 A-142594 
                 AGCCCCAGCGUG 
                 1199 
                 UCCUGAGCAC 
                 1407 
               
               
                   
                   
                 UGGGGCUdTdT 
                   
                   
                 CUCAGGAdTdT 
                   
                 GCUGGGGCU 
                   
               
               
                   
               
               
                 AD-71108 
                 A-142595 
                 CUGGGGCUGCGA 
                 992 
                 A-142596 
                 UCGAGGGUCGCA 
                 1200 
                 CUGGGGCUGC 
                 1408 
               
               
                   
                   
                 CCCUCGAdTdT 
                   
                   
                 GCCCCAGdTdT 
                   
                 GACCCUCGA 
                   
               
               
                   
               
               
                 AD-71109 
                 A-142597 
                 CGACCCUCGACC 
                 993 
                 A-142598 
                 AGUCGGUGGUCG 
                 1201 
                 CGACCCUCGA 
                 1409 
               
               
                   
                   
                 ACCGACUdTdT 
                   
                   
                 AGGGUCGdTdT 
                   
                 CCACCGACU 
                   
               
               
                   
               
               
                 AD-71110 
                 A-142599 
                 CACCGACUGCGA 
                 994 
                 A-142600 
                 UACGAUGUCGCA 
                 1202 
                 CACCGACUGC 
                 1410 
               
               
                   
                   
                 CAUCGUAdTdT 
                   
                   
                 GUCGGUGdTdT 
                   
                 GACAUCGUG 
                   
               
               
                   
               
               
                 AD-71111 
                 A-142601 
                 CAUCGUGCGCCG 
                 995 
                 A-142602 
                 UCAGGCGCGGCG 
                 1203 
                 CAUCGUGCGC 
                 1411 
               
               
                   
                   
                 CGCCUGAdTdT 
                   
                   
                 CACGAUGdTdT 
                   
                 CGCGCCUGC 
                   
               
               
                   
               
               
                 AD-71112 
                 A-142603 
                 CGCGCCUGCGAG 
                 996 
                 A-142604 
                 ACACGCUCUCGC 
                 1204 
                 CGCGCCUGCG 
                 1412 
               
               
                   
                   
                 AGCGUGUdTdT 
                   
                   
                 AGGCGCGdTdT 
                   
                 AGAGCGUGU 
                   
               
               
                   
               
               
                 AD-71113 
                 A-142605 
                 AGCGUGUCUACG 
                 997 
                 A-142606 
                 UAGCGCGCGUAG 
                 1205 
                 AGCGUGUCUA 
                 1413 
               
               
                   
                   
                 CGCGCUAdTdT 
                   
                   
                 ACACGCUdTdT 
                   
                 CGCGCGCUG 
                   
               
               
                   
               
               
                 AD-71114 
                 A-142607 
                 CGCGCUGCGCAC 
                 998 
                 A-142608 
                 AGCACAUGUGCG 
                 1206 
                 CGCGCUGCGC 
                 1414 
               
               
                   
                   
                 AUGUGCUdTdT 
                   
                   
                 CAGCGCGdTdT 
                   
                 ACAUGUGCU 
                   
               
               
                   
               
               
                 AD-71115 
                 A-142609 
                 ACAUGUGCUCGG 
                 999 
                 A-142610 
                 AGCCCCGCCGAG 
                 1207 
                 ACAUGUGCUC 
                 1415 
               
               
                   
                   
                 CGGGGCUdTdT 
                   
                   
                 CACAUGUdTdT 
                   
                 GGCGGGGCU 
                   
               
               
                   
               
               
                 AD-71116 
                 A-142611 
                 CGGGGCUGGCGG 
                 1000 
                 A-142612 
                 AUGACGCCCGCC 
                 1208 
                 CGGGGCUGGC 
                 1416 
               
               
                   
                   
                 GCGUCAUdTdT 
                   
                   
                 AGCCCCGdTdT 
                   
                 GGGCGUCAU 
                   
               
               
                   
               
               
                 AD-71117 
                 A-142613 
                 CGGGCGUCAUCA 
                 1001 
                 A-142614 
                 AUGCGGUUGAUG 
                 1209 
                 CGGGCGUCAU 
                 1417 
               
               
                   
                   
                 ACCGCAUdTdT 
                   
                   
                 ACGCCCGdTdT 
                   
                 CAACCGCAU 
                   
               
               
                   
               
               
                 AD-71118 
                 A-142615 
                 AACCGCAUGCGC 
                 1002 
                 A-142616 
                 UGCUCUCGCGCA 
                 1210 
                 AACCGCAUGC 
                 1418 
               
               
                   
                   
                 GAGAGCAdTdT 
                   
                   
                 UGCGGUUdTdT 
                   
                 GCGAGAGCC 
                   
               
               
                   
               
               
                 AD-71119 
                 A-142617 
                 AGAGCCGCAGCG 
                 1003 
                 A-142618 
                 ACGUCCUCGCUG 
                 1211 
                 AGAGCCGCAG 
                 1419 
               
               
                   
                   
                 AGGACGUdTdT 
                   
                   
                 CGGCUCUdTdT 
                   
                 CGAGGACGU 
                   
               
               
                   
               
               
                 AD-71120 
                 A-142619 
                 CGAGGACGUAAU 
                 1004 
                 A-142620 
                 UAUGCGCAUUAC 
                 1212 
                 CGAGGACGUA 
                 1420 
               
               
                   
                   
                 GCGCAUAdTdT 
                   
                   
                 GUCCUCGdTdT 
                   
                 AUGCGCAUC 
                   
               
               
                   
               
               
                 AD-71121 
                 A-142621 
                 UGCGCAUCACUG 
                 1005 
                 A-142622 
                 ACGCCCACAGUG 
                 1213 
                 UGCGCAUCAC 
                 1421 
               
               
                   
                   
                 UGGGCGUdTdT 
                   
                   
                 AUGCGCAdTdT 
                   
                 UGUGGGCGU 
                   
               
               
                   
               
               
                 AD-71122 
                 A-142623 
                 UGGGCGUGGAUG 
                 1006 
                 A-142624 
                 ACGGAGCCAUCC 
                 1214 
                 UGGGCGUGGA 
                 1422 
               
               
                   
                   
                 GCUCCGUdTdT 
                   
                   
                 ACGCCCAdTdT 
                   
                 UGGCUCCGU 
                   
               
               
                   
               
               
                 AD-71123 
                 A-142625 
                 UGGCUCCGUGUA 
                 1007 
                 A-142626 
                 UAGCUUGUACAC 
                 1215 
                 UGGCUCCGUG 
                 1423 
               
               
                   
                   
                 CAAGCUAdTdT 
                   
                   
                 GGAGCCAdTdT 
                   
                 UACAAGCUG 
                   
               
               
                   
               
               
                 AD-71124 
                 A-142627 
                 UACAAGCUGCAC 
                 1008 
                 A-142628 
                 AGCUGGGGUGCA 
                 1216 
                 UACAAGCUGC 
                 1424 
               
               
                   
                   
                 CCCAGCUdTdT 
                   
                   
                 GCUUGUAdTdT 
                   
                 ACCCCAGCU 
                   
               
               
                   
               
               
                 AD-71125 
                 A-142629 
                 CCAGCUUCAAGG 
                 1009 
                 A-142630 
                 AACCGCUCCUUG 
                 1217 
                 CCAGCUUCAA 
                 1425 
               
               
                   
                   
                 AGCGGUUdTdT 
                   
                   
                 AAGCUGGdTdT 
                   
                 GGAGCGGUU 
                   
               
               
                   
               
               
                 AD-71126 
                 A-142631 
                 AGGAGCGGUUCC 
                 1010 
                 A-142632 
                 UUGGCAUGGAAC 
                 1218 
                 AGGAGCGGUU 
                 1426 
               
               
                   
                   
                 AUGCCAAdTdT 
                   
                   
                 CGCUCCUdTdT 
                   
                 CCAUGCCAG 
                   
               
               
                   
               
               
                 AD-71127 
                 A-142633 
                 AUGCCAGCGUGC 
                 1011 
                 A-142634 
                 AGCCUGCGCACG 
                 1219 
                 AUGCCAGCGU 
                 1427 
               
               
                   
                   
                 GCAGGCUdTdT 
                   
                   
                 CUGGCAUdTdT 
                   
                 GCGCAGGCU 
                   
               
               
                   
               
               
                 AD-71128 
                 A-142635 
                 CGCAGGCUGACG 
                 1012 
                 A-142636 
                 AGCUGGGCGUCA 
                 1220 
                 CGCAGGCUGA 
                 1428 
               
               
                   
                   
                 CCCAGCUdTdT 
                   
                   
                 GCCUGCGdTdT 
                   
                 CGCCCAGCU 
                   
               
               
                   
               
               
                 AD-71129 
                 A-142637 
                 CCCAGCUGCGAG 
                 1013 
                 A-142638 
                 AGGUGAUCUCGC 
                 1221 
                 CCCAGCUGCG 
                 1429 
               
               
                   
                   
                 AUCACCUdTdT 
                   
                   
                 AGCUGGGdTdT 
                   
                 AGAUCACCU 
                   
               
               
                   
               
               
                 AD-71130 
                 A-142639 
                 GAGAUCACCUUC 
                 1014 
                 A-142640 
                 ACUCGAUGAAGG 
                 1222 
                 GAGAUCACCU 
                 1430 
               
               
                   
                   
                 AUCGAGUdTdT 
                   
                   
                 UGAUCUCdTdT 
                   
                 UCAUCGAGU 
                   
               
               
                   
               
               
                 AD-71131 
                 A-142641 
                 AUCGAGUCGGAG 
                 1015 
                 A-142642 
                 UGCCCUCCUCCG 
                 1223 
                 AUCGAGUCGG 
                 1431 
               
               
                   
                   
                 GAGGGCAdTdT 
                   
                   
                 ACUCGAUdTdT 
                   
                 AGGAGGGCA 
                   
               
               
                   
               
               
                 AD-71132 
                 A-142643 
                 AGGAGGGCAGUG 
                 1016 
                 A-142644 
                 UCCCGGCCACUG 
                 1224 
                 AGGAGGGCAG 
                 1432 
               
               
                   
                   
                 GCCGGGAdTdT 
                   
                   
                 CCCUCCUdTdT 
                   
                 UGGCCGGGG 
                   
               
               
                   
               
               
                 AD-71133 
                 A-142645 
                 CCGGGGCGCGGC 
                 1017 
                 A-142646 
                 UACCAGGGCCGC 
                 1225 
                 CCGGGGCGCG 
                 1433 
               
               
                   
                   
                 CCUGGUAdTdT 
                   
                   
                 GCCCCGGdTdT 
                   
                 GCCCUGGUC 
                   
               
               
                   
               
               
                 AD-71134 
                 A-142647 
                 CCCUGGUCUCGG 
                 1018 
                 A-142648 
                 UCCACCGCCGAG 
                 1226 
                 CCCUGGUCUC 
                 1434 
               
               
                   
                   
                 CGGUGGAdTdT 
                   
                   
                 ACCAGGGdTdT 
                   
                 GGCGGUGGC 
                   
               
               
                   
               
               
                 AD-71135 
                 A-142649 
                 GCGGUGGCCUGU 
                 1019 
                 A-142650 
                 UCUUCUUACAGG 
                 1227 
                 GCGGUGGCCU 
                 1435 
               
               
                   
                   
                 AAGAAGAdTdT 
                   
                   
                 CCACCGCdTdT 
                   
                 GUAAGAAGG 
                   
               
               
                   
               
               
                 AD-71136 
                 A-142651 
                 UAAGAAGGCCUG 
                 1020 
                 A-142652 
                 UAGCAUACAGGC 
                 1228 
                 UAAGAAGGCC 
                 1436 
               
               
                   
                   
                 UAUGCUAdTdT 
                   
                   
                 CUUCUUAdTdT 
                   
                 UGUAUGCUG 
                   
               
               
                   
               
               
                 AD-71137 
                 A-142653 
                 CUGUAUGCUGGG 
                 1021 
                 A-142654 
                 UCACUGGCCCAG 
                 1229 
                 CUGUAUGCUG 
                 1437 
               
               
                   
                   
                 CCAGUGAdTdT 
                   
                   
                 CAUACAGdTdT 
                   
                 GGCCAGUGA 
                   
               
               
                   
               
               
                 AD-71138 
                 A-142655 
                 CAGUGAGAGCAG 
                 1022 
                 A-142656 
                 UCGGCCACUGCU 
                 1230 
                 CAGUGAGAGC 
                 1438 
               
               
                   
                   
                 UGGCCGAdTdT 
                   
                   
                 CUCACUGdTdT 
                   
                 AGUGGCCGC 
                   
               
               
                   
               
               
                 AD-71139 
                 A-142657 
                 CAGUGGCCGCAA 
                 1023 
                 A-142658 
                 UCUGCGCUUGCG 
                 1231 
                 CAGUGGCCGC 
                 1439 
               
               
                   
                   
                 GCGCAGAdTdT 
                   
                   
                 GCCACUGdTdT 
                   
                 AAGCGCAGG 
                   
               
               
                   
               
               
                 AD-71140 
                 A-142659 
                 AGCGCAGGGAGG 
                 1024 
                 A-142660 
                 UUGGCAUCCUCC 
                 1232 
                 AGCGCAGGGA 
                 1440 
               
               
                   
                   
                 AUGCCAAdTdT 
                   
                   
                 CUGCGCUdTdT 
                   
                 GGAUGCCAC 
                   
               
               
                   
               
               
                 AD-71141 
                 A-142661 
                 UGCCACAGCCCC 
                 1025 
                 A-142662 
                 UUGCUGUGGGGC 
                 1233 
                 UGCCACAGCC 
                 1441 
               
               
                   
                   
                 ACAGCAAdTdT 
                   
                   
                 UGUGGCAdTdT 
                   
                 CCACAGCAC 
                   
               
               
                   
               
               
                 AD-71142 
                 A-142663 
                 CACAGCACCCAG 
                 1026 
                 A-142664 
                 AUGGAGCCUGGG 
                 1234 
                 CACAGCACCC 
                 1442 
               
               
                   
                   
                 GCUCCAUdTdT 
                   
                   
                 UGCUGUGdTdT 
                   
                 AGGCUCCAU 
                   
               
               
                   
               
               
                 AD-71143 
                 A-142665 
                 AGGCUCCAUGGG 
                 1027 
                 A-142666 
                 UCACUUCCCCAU 
                 1235 
                 AGGCUCCAUG 
                 1443 
               
               
                   
                   
                 GAAGUGAdTdT 
                   
                   
                 GGAGCCUdTdT 
                   
                 GGGAAGUGC 
                   
               
               
                   
               
               
                 AD-71144 
                 A-142667 
                 GGAAGUGCUCCC 
                 1028 
                 A-142668 
                 ACGUGUGGGGAG 
                 1236 
                 GGAAGUGCUC 
                 1444 
               
               
                   
                   
                 CACACGUdTdT 
                   
                   
                 CACUUCCdTdT 
                   
                 CCCACACGU 
                   
               
               
                   
               
               
                 AD-71145 
                 A-142669 
                 CCACACGUGCUC 
                 1029 
                 A-142670 
                 AGGCUGCGAGCA 
                 1237 
                 CCACACGUGC 
                 1445 
               
               
                   
                   
                 GCAGCCUdTdT 
                   
                   
                 CGUGUGGdTdT 
                   
                 UCGCAGCCU 
                   
               
               
                   
               
               
                 AD-71146 
                 A-142671 
                 UCGCAGCCUGGC 
                 1030 
                 A-142672 
                 UUGCCCCGCCAG 
                 1238 
                 UCGCAGCCUG 
                 1446 
               
               
                   
                   
                 GGGGCAAdTdT 
                   
                   
                 GCUGCGAdTdT 
                   
                 GCGGGGCAG 
                   
               
               
                   
               
               
                 AD-71147 
                 A-142673 
                 CGGGGCAGGAGG 
                 1031 
                 A-142674 
                 UGCCAGGCCUCC 
                 1239 
                 CGGGGCAGGA 
                 1447 
               
               
                   
                   
                 CCUGGCAdTdT 
                   
                   
                 UGCCCCGdTdT 
                   
                 GGCCUGGCC 
                   
               
               
                   
               
               
                 AD-71148 
                 A-142675 
                 CCUGGCCUUGUC 
                 1032 
                 A-142676 
                 UGGUCCUGACAA 
                 1240 
                 CCUGGCCUUG 
                 1448 
               
               
                   
                   
                 AGGACCAdTdT 
                   
                   
                 GGCCAGGdTdT 
                   
                 UCAGGACCC 
                   
               
               
                   
               
               
                 AD-71149 
                 A-142677 
                 CAGGACCCAGGC 
                 1033 
                 A-142678 
                 UCAGGCGGCCUG 
                 1241 
                 CAGGACCCAG 
                 1449 
               
               
                   
                   
                 CGCCUGAdTdT 
                   
                   
                 GGUCCUGdTdT 
                   
                 GCCGCCUGC 
                   
               
               
                   
               
               
                 AD-71150 
                 A-142679 
                 CCGCCUGCCAUA 
                 1034 
                 A-142680 
                 UCAGCGGUAUGG 
                 1242 
                 CCGCCUGCCA 
                 1450 
               
               
                   
                   
                 CCGCUGAdTdT 
                   
                   
                 CAGGCGGdTdT 
                   
                 UACCGCUGG 
                   
               
               
                   
               
               
                 AD-71151 
                 A-142681 
                 UACCGCUGGGGA 
                 1035 
                 A-142682 
                 UCUCUGUUCCCC 
                 1243 
                 UACCGCUGGG 
                 1451 
               
               
                   
                   
                 ACAGAGAdTdT 
                   
                   
                 AGCGGUAdTdT 
                   
                 GAACAGAGC 
                   
               
               
                   
               
               
                 AD-71152 
                 A-142683 
                 AACAGAGCGGGC 
                 1036 
                 A-142684 
                 UGAAGAGGCCCG 
                 1244 
                 AACAGAGCGG 
                 1452 
               
               
                   
                   
                 CUCUUCAdTdT 
                   
                   
                 CUCUGUUdTdT 
                   
                 GCCUCUUCC 
                   
               
               
                   
               
               
                 AD-71153 
                 A-142685 
                 CUCUUCCCUCAG 
                 1037 
                 A-142686 
                 UGAAAAACUGAG 
                 1245 
                 CUCUUCCCUC 
                 1453 
               
               
                   
                   
                 UUUUUCAdTdT 
                   
                   
                 GGAAGAGdTdT 
                   
                 AGUUUUUCG 
                   
               
               
                   
               
               
                 AD-71154 
                 A-142687 
                 UUUUUCGGUGGG 
                 1038 
                 A-142688 
                 UGGCUGUCCCAC 
                 1246 
                 UUUUUCGGUG 
                 1454 
               
               
                   
                   
                 ACAGCCAdTdT 
                   
                   
                 CGAAAAAdTdT 
                   
                 GGACAGCCC 
                   
               
               
                   
               
               
                 AD-71155 
                 A-142689 
                 GGGACAGCCCCA 
                 1039 
                 A-142690 
                 AGGGCCCUGGGG 
                 1247 
                 GGGACAGCCC 
                 1455 
               
               
                   
                   
                 GGGCCCUdTdT 
                   
                   
                 CUGUCCCdTdT 
                   
                 CAGGGCCCU 
                   
               
               
                   
               
               
                 AD-71156 
                 A-142691 
                 AGGGCCCUAACG 
                 1040 
                 A-142692 
                 UCACCCCCGUUA 
                 1248 
                 AGGGCCCUAA 
                 1456 
               
               
                   
                   
                 GGGGUGAdTdT 
                   
                   
                 GGGCCCUdTdT 
                   
                 CGGGGGUGC 
                   
               
               
                   
               
               
                 AD-71157 
                 A-142693 
                 GGGUGCGGCAGG 
                 1041 
                 A-142694 
                 UCCUGCUCCUGC 
                 1249 
                 GGGUGCGGCA 
                 1457 
               
               
                   
                   
                 AGCAGGAdTdT 
                   
                   
                 CGCACCCdTdT 
                   
                 GGAGCAGGA 
                   
               
               
                   
               
               
                 AD-71158 
                 A-142695 
                 AGGAGCAGGAAC 
                 1042 
                 A-142696 
                 AGUCUCUGUUCC 
                 1250 
                 AGGAGCAGGA 
                 1458 
               
               
                   
                   
                 AGAGACUdTdT 
                   
                   
                 UGCUCCUdTdT 
                   
                 ACAGAGACU 
                   
               
               
                   
               
               
                 AD-71159 
                 A-142697 
                 AGAGACUCUGGA 
                 1043 
                 A-142698 
                 UGGGGCUUCCAG 
                 1251 
                 AGAGACUCUG 
                 1459 
               
               
                   
                   
                 AGCCCCAdTdT 
                   
                   
                 AGUCUCUdTdT 
                   
                 GAAGCCCCC 
                   
               
               
                   
               
               
                 AD-71160 
                 A-142699 
                 AAGCCCCCCACC 
                 1044 
                 A-142700 
                 UGAGAAAGGUGG 
                 1252 
                 AAGCCCCCCA 
                 1460 
               
               
                   
                   
                 UUUCUCAdTdT 
                   
                   
                 GGGGCUUdTdT 
                   
                 CCUUUCUCG 
                   
               
               
                   
               
               
                 AD-71161 
                 A-142701 
                 UUUCUCGCUGGA 
                 1045 
                 A-142702 
                 AAUUGAUUCCAG 
                 1253 
                 UUUCUCGCUG 
                 1461 
               
               
                   
                   
                 AUCAAUUdTdT 
                   
                   
                 CGAGAAAdTdT 
                   
                 GAAUCAAUU 
                   
               
               
                   
               
               
                 AD-71162 
                 A-142703 
                 AAUCAAUUUCCC 
                 1046 
                 A-142704 
                 UCCUUCUGGGAA 
                 1254 
                 AAUCAAUUUC 
                 1462 
               
               
                   
                   
                 AGAAGGAdTdT 
                   
                   
                 AUUGAUUdTdT 
                   
                 CCAGAAGGG 
                   
               
               
                   
               
               
                 AD-71163 
                 A-142705 
                 CCCAGAAGGGAG 
                 1047 
                 A-142706 
                 UGAGCAACUCCC 
                 1255 
                 CCCAGAAGGG 
                 1463 
               
               
                   
                   
                 UUGCUCAdTdT 
                   
                   
                 UUCUGGGdTdT 
                   
                 AGUUGCUCA 
                   
               
               
                   
               
               
                 AD-71164 
                 A-142707 
                 UUGCUCACUCAG 
                 1048 
                 A-142708 
                 UAAAGUCCUGAG 
                 1256 
                 UUGCUCACUC 
                 1464 
               
               
                   
                   
                 GACUUUAdTdT 
                   
                   
                 UGAGCAAdTdT 
                   
                 AGGACUUUG 
                   
               
               
                   
               
               
                 AD-71165 
                 A-142709 
                 AGGACUUUGAUG 
                 1049 
                 A-142710 
                 UGAAAUGCAUCA 
                 1257 
                 AGGACUUUGA 
                 1465 
               
               
                   
                   
                 CAUUUCAdTdT 
                   
                   
                 AAGUCCUdTdT 
                   
                 UGCAUUUCC 
                   
               
               
                   
               
               
                 AD-71166 
                 A-142711 
                 AUUUCCACACUG 
                 1050 
                 A-142712 
                 UCUCUGACAGUG 
                 1258 
                 AUUUCCACAC 
                 1466 
               
               
                   
                   
                 UCAGAGAdTdT 
                   
                   
                 UGGAAAUdTdT 
                   
                 UGUCAGAGC 
                   
               
               
                   
               
               
                 AD-71167 
                 A-142713 
                 UGUCAGAGCUGU 
                 1051 
                 A-142714 
                 UAGGCCAACAGC 
                 1259 
                 UGUCAGAGCU 
                 1467 
               
               
                   
                   
                 UGGCCUAdTdT 
                   
                   
                 UCUGACAdTdT 
                   
                 GUUGGCCUC 
                   
               
               
                   
               
               
                 AD-71168 
                 A-142715 
                 UUGGCCUCGCCU 
                 1052 
                 A-142716 
                 UGGGCCCAGGCG 
                 1260 
                 UUGGCCUCGC 
                 1468 
               
               
                   
                   
                 GGGCCCAdTdT 
                   
                   
                 AGGCCAAdTdT 
                   
                 CUGGGCCCA 
                   
               
               
                   
               
               
                 AD-71169 
                 A-142717 
                 CUGGGCCCAGGC 
                 1053 
                 A-142718 
                 UCCCAGAGCCUG 
                 1261 
                 CUGGGCCCAG 
                 1469 
               
               
                   
                   
                 UCUGGGAdTdT 
                   
                   
                 GGCCCAGdTdT 
                   
                 GCUCUGGGA 
                   
               
               
                   
               
               
                 AD-71170 
                 A-142719 
                 CUCUGGGAAGGG 
                 1054 
                 A-142720 
                 AGGGCACCCCUU 
                 1262 
                 CUCUGGGAAG 
                 1470 
               
               
                   
                   
                 GUGCCCUdTdT 
                   
                   
                 CCCAGAGdTdT 
                   
                 GGGUGCCCU 
                   
               
               
                   
               
               
                 AD-71171 
                 A-142721 
                 UGCCCUCUGGAU 
                 1055 
                 A-142722 
                 UAGCAGGAUCCA 
                 1263 
                 UGCCCUCUGG 
                 1471 
               
               
                   
                   
                 CCUGCUAdTdT 
                   
                   
                 GAGGGCAdTdT 
                   
                 AUCCUGCUG 
                   
               
               
                   
               
               
                 AD-71172 
                 A-142723 
                 UCCUGCUGUGGC 
                 1056 
                 A-142724 
                 AAGUGAGGCCAC 
                 1264 
                 UCCUGCUGUG 
                 1472 
               
               
                   
                   
                 CUCACUUdTdT 
                   
                   
                 AGCAGGAdTdT 
                   
                 GCCUCACUU 
                   
               
               
                   
               
               
                 AD-71173 
                 A-142725 
                 CCUCACUUCCCU 
                 1057 
                 A-142726 
                 AGUUCCCAGGGA 
                 1265 
                 CCUCACUUCC 
                 1473 
               
               
                   
                   
                 GGGAACUdTdT 
                   
                   
                 AGUGAGGdTdT 
                   
                 CUGGGAACU 
                   
               
               
                   
               
               
                 AD-71174 
                 A-142727 
                 CUGGGAACUCAU 
                 1058 
                 A-142728 
                 ACACAGGAUGAG 
                 1266 
                 CUGGGAACUC 
                 1474 
               
               
                   
                   
                 CCUGUGUdTdT 
                   
                   
                 UUCCCAGdTdT 
                   
                 AUCCUGUGU 
                   
               
               
                   
               
               
                 AD-71175 
                 A-142729 
                 CCUGUGUGGGGA 
                 1059 
                 A-142730 
                 AGCUGCCUCCCC 
                 1267 
                 CCUGUGUGGG 
                 1475 
               
               
                   
                   
                 GGCAGCUdTdT 
                   
                   
                 ACACAGGdTdT 
                   
                 GAGGCAGCU 
                   
               
               
                   
               
               
                 AD-71176 
                 A-142731 
                 GGAGGCAGCUCC 
                 1060 
                 A-142732 
                 AGCUGUUGGAGC 
                 1268 
                 GGAGGCAGCU 
                 1476 
               
               
                   
                   
                 AACAGCUdTdT 
                   
                   
                 UGCCUCCdTdT 
                   
                 CCAACAGCU 
                   
               
               
                   
               
               
                 AD-71177 
                 A-142733 
                 CAACAGCUUGAC 
                 1061 
                 A-142734 
                 AGGUCUGGUCAA 
                 1269 
                 CAACAGCUUG 
                 1477 
               
               
                   
                   
                 CAGACCUdTdT 
                   
                   
                 GCUGUUGdTdT 
                   
                 ACCAGACCU 
                   
               
               
                   
               
               
                 AD-71178 
                 A-142735 
                 CCAGACCUAGAC 
                 1062 
                 A-142736 
                 UGCCCAGGUCUA 
                 1270 
                 CCAGACCUAG 
                 1478 
               
               
                   
                   
                 CUGGGCAdTdT 
                   
                   
                 GGUCUGGdTdT 
                   
                 ACCUGGGCC 
                   
               
               
                   
               
               
                 AD-71179 
                 A-142737 
                 CUGGGCCAAAAG 
                 1063 
                 A-142738 
                 UGCUGCCCUUUU 
                 1271 
                 CUGGGCCAAA 
                 1479 
               
               
                   
                   
                 GGCAGCAdTdT 
                   
                   
                 GGCCCAGdTdT 
                   
                 AGGGCAGCC 
                   
               
               
                   
               
               
                 AD-71180 
                 A-142739 
                 AGGGCAGCCAGG 
                 1064 
                 A-142740 
                 AGCAGCCCCUGG 
                 1272 
                 AGGGCAGCCA 
                 1480 
               
               
                   
                   
                 GGCUGCUdTdT 
                   
                   
                 CUGCCCUdTdT 
                   
                 GGGGCUGCU 
                   
               
               
                   
               
               
                 AD-71181 
                 A-142741 
                 GGGCUGCUCAUC 
                 1065 
                 A-142742 
                 ACUGGGUGAUGA 
                 1273 
                 GGGCUGCUCA 
                 1481 
               
               
                   
                   
                 ACCCAGUdTdT 
                   
                   
                 GCAGCCCdTdT 
                   
                 UCACCCAGU 
                   
               
               
                   
               
               
                 AD-71182 
                 A-142743 
                 ACCCAGUCCUGG 
                 1066 
                 A-142744 
                 AAAAUGGCCAGG 
                 1274 
                 ACCCAGUCCU 
                 1482 
               
               
                   
                   
                 CCAUUUUdTdT 
                   
                   
                 ACUGGGUdTdT 
                   
                 GGCCAUUUU 
                   
               
               
                   
               
               
                 AD-71183 
                 A-142745 
                 GCCAUUUUCUUG 
                 1067 
                 A-142746 
                 UCUCAGGCAAGA 
                 1275 
                 GCCAUUUUCU 
                 1483 
               
               
                   
                   
                 CCUGAGAdTdT 
                   
                   
                 AAAUGGCdTdT 
                   
                 UGCCUGAGG 
                   
               
               
                   
               
               
                 AD-71184 
                 A-142747 
                 CCUGAGGCUCAA 
                 1068 
                 A-142748 
                 UGGCCUCUUGAG 
                 1276 
                 CCUGAGGCUC 
                 1484 
               
               
                   
                   
                 GAGGCCAdTdT 
                   
                   
                 CCUCAGGdTdT 
                   
                 AAGAGGCCC 
                   
               
               
                   
               
               
                 AD-71185 
                 A-142749 
                 AAGAGGCCCAGG 
                 1069 
                 A-142750 
                 AUUGCUCCCUGG 
                 1277 
                 AAGAGGCCCA 
                 1485 
               
               
                   
                   
                 GAGCAAUdTdT 
                   
                   
                 GCCUCUUdTdT 
                   
                 GGGAGCAAU 
                   
               
               
                   
               
               
                 AD-71186 
                 A-142751 
                 GGAGCAAUGGGA 
                 1070 
                 A-142752 
                 AGCCCCCUCCCA 
                 1278 
                 GGAGCAAUGG 
                 1486 
               
               
                   
                   
                 GGGGGCUdTdT 
                   
                   
                 UUGCUCCdTdT 
                   
                 GAGGGGGCU 
                   
               
               
                   
               
               
                 AD-71187 
                 A-142753 
                 AGGGGGCUCCAU 
                 1071 
                 A-142754 
                 UUCCUCCAUGGA 
                 1279 
                 AGGGGGCUCC 
                 1487 
               
               
                   
                   
                 GGAGGAAdTdT 
                   
                   
                 GCCCCCUdTdT 
                   
                 AUGGAGGAG 
                   
               
               
                   
               
               
                 AD-71188 
                 A-142755 
                 GGAGGAGGUGUC 
                 1072 
                 A-142756 
                 AGCUUGGGACAC 
                 1280 
                 GGAGGAGGUG 
                 1488 
               
               
                   
                   
                 CCAAGCUdTdT 
                   
                   
                 CUCCUCCdTdT 
                   
                 UCCCAAGCU 
                   
               
               
                   
               
               
                 AD-71189 
                 A-142757 
                 UCCCAAGCUUUG 
                 1073 
                 A-142758 
                 UGGUAUUCAAAG 
                 1281 
                 UCCCAAGCUU 
                 1489 
               
               
                   
                   
                 AAUACCAdTdT 
                   
                   
                 CUUGGGAdTdT 
                   
                 UGAAUACCC 
                   
               
               
                   
               
               
                 AD-71190 
                 A-142759 
                 AAUACCCCCAGA 
                 1074 
                 A-142760 
                 AAAGGUCUCUGG 
                 1282 
                 AAUACCCCCA 
                 1490 
               
               
                   
                   
                 GACCUUUdTdT 
                   
                   
                 GGGUAUUdTdT 
                   
                 GAGACCUUU 
                   
               
               
                   
               
               
                 AD-71191 
                 A-142761 
                 AGAGACCUUUUC 
                 1075 
                 A-142762 
                 UGGGAGAGAAAA 
                 1283 
                 AGAGACCUUU 
                 1491 
               
               
                   
                   
                 UCUCCCAdTdT 
                   
                   
                 GGUCUCUdTdT 
                   
                 UCUCUCCCA 
                   
               
               
                   
               
               
                 AD-71192 
                 A-142763 
                 UCUCCCAUACCA 
                 1076 
                 A-142764 
                 UCAGUGAUGGUA 
                 1284 
                 UCUCCCAUAC 
                 1492 
               
               
                   
                   
                 UCACUGAdTdT 
                   
                   
                 UGGGAGAdTdT 
                   
                 CAUCACUGA 
                   
               
               
                   
               
               
                 AD-71193 
                 A-142765 
                 UCACUGAGUGGC 
                 1077 
                 A-142766 
                 AUCACAAGCCAC 
                 1285 
                 UCACUGAGUG 
                 1493 
               
               
                   
                   
                 UUGUGAUdTdT 
                   
                   
                 UCAGUGAdTdT 
                   
                 GCUUGUGAU 
                   
               
               
                   
               
               
                 AD-71194 
                 A-142767 
                 GGCUUGUGAUUC 
                 1078 
                 A-142768 
                 UAUCCCAGAAUC 
                 1286 
                 GGCUUGUGAU 
                 1494 
               
               
                   
                   
                 UGGGAUAdTdT 
                   
                   
                 ACAAGCCdTdT 
                   
                 UCUGGGAUG 
                   
               
               
                   
               
               
                 AD-71195 
                 A-142769 
                 UGGGAUGGACCC 
                 1079 
                 A-142770 
                 UCUGCGAGGGUC 
                 1287 
                 UGGGAUGGAC 
                 1495 
               
               
                   
                   
                 UCGCAGAdTdT 
                   
                   
                 CAUCCCAdTdT 
                   
                 CCUCGCAGC 
                   
               
               
                   
               
               
                 AD-71196 
                 A-142771 
                 UCGCAGCAGGUG 
                 1080 
                 A-142772 
                 UCUCUUGCACCU 
                 1288 
                 UCGCAGCAGG 
                 1496 
               
               
                   
                   
                 CAAGAGAdTdT 
                   
                   
                 GCUGCGAdTdT 
                   
                 UGCAAGAGA 
                   
               
               
                   
               
               
                 AD-71197 
                 A-142773 
                 UGCAAGAGACAG 
                 1081 
                 A-142774 
                 UGGGGCUCUGUC 
                 1289 
                 UGCAAGAGAC 
                 1497 
               
               
                   
                   
                 AGCCCCAdTdT 
                   
                   
                 UCUUGCAdTdT 
                   
                 AGAGCCCCC 
                   
               
               
                   
               
               
                 AD-71198 
                 A-142775 
                 AGAGCCCCCAAG 
                 1082 
                 A-142776 
                 UCAGAGGCUUGG 
                 1290 
                 AGAGCCCCCA 
                 1498 
               
               
                   
                   
                 CCUCUGAdTdT 
                   
                   
                 GGGCUCUdTdT 
                   
                 AGCCUCUGC 
                   
               
               
                   
               
               
                 AD-71199 
                 A-142777 
                 CUCUGCCCCAAG 
                 1083 
                 A-142778 
                 UGGGCCCCUUGG 
                 1291 
                 CUCUGCCCCA 
                 1499 
               
               
                   
                   
                 GGGCCCAdTdT 
                   
                   
                 GGCAGAGdTdT 
                   
                 AGGGGCCCA 
                   
               
               
                   
               
               
                 AD-71200 
                 A-142779 
                 AAGGGGCCCACA 
                 1084 
                 A-142780 
                 UCCCCUUUGUGG 
                 1292 
                 AAGGGGCCCA 
                 1500 
               
               
                   
                   
                 AAGGGGAdTdT 
                   
                   
                 GCCCCUUdTdT 
                   
                 CAAAGGGGA 
                   
               
               
                   
               
               
                 AD-71201 
                 A-142781 
                 AAAGGGGAGAAG 
                 1085 
                 A-142782 
                 UCUGGCCCUUCU 
                 1293 
                 AAAGGGGAGA 
                 1501 
               
               
                   
                   
                 GGCCAGAdTdT 
                   
                   
                 CCCCUUUdTdT 
                   
                 AGGGCCAGC 
                   
               
               
                   
               
               
                 AD-71202 
                 A-142783 
                 GGGCCAGCCCUA 
                 1086 
                 A-142784 
                 UAAGAUGUAGGG 
                 1294 
                 GGGCCAGCCC 
                 1502 
               
               
                   
                   
                 CAUCUUAdTdT 
                   
                   
                 CUGGCCCdTdT 
                   
                 UACAUCUUC 
                   
               
               
                   
               
               
                 AD-71203 
                 A-142785 
                 AUCUUCAGCUCC 
                 1087 
                 A-142786 
                 UGCUAUGGGAGC 
                 1295 
                 AUCUUCAGCU 
                 1503 
               
               
                   
                   
                 CAUAGCAdTdT 
                   
                   
                 UGAAGAUdTdT 
                   
                 CCCAUAGCG 
                   
               
               
                   
               
               
                 AD-71204 
                 A-142787 
                 UCCCAUAGCGCU 
                 1088 
                 A-142788 
                 UUGAGCCAGCGC 
                 1296 
                 UCCCAUAGCG 
                 1504 
               
               
                   
                   
                 GGCUCAAdTdT 
                   
                   
                 UAUGGGAdTdT 
                   
                 CUGGCUCAG 
                   
               
               
                   
               
               
                 AD-71205 
                 A-142789 
                 UGGCUCAGGAAG 
                 1089 
                 A-142790 
                 UGGGUUUCUUCC 
                 1297 
                 UGGCUCAGGA 
                 1505 
               
               
                   
                   
                 AAACCCAdTdT 
                   
                   
                 UGAGCCAdTdT 
                   
                 AGAAACCCC 
                   
               
               
                   
               
               
                 AD-71206 
                 A-142791 
                 AACCCCAAGCAG 
                 1090 
                 A-142792 
                 UUGAAUGCUGCU 
                 1298 
                 AACCCCAAGC 
                 1506 
               
               
                   
                   
                 CAUUCAAdTdT 
                   
                   
                 UGGGGUUdTdT 
                   
                 AGCAUUCAG 
                   
               
               
                   
               
               
                 AD-71207 
                 A-142793 
                 CAGCAUUCAGCA 
                 1091 
                 A-142794 
                 UGGGGUGUGCUG 
                 1299 
                 CAGCAUUCAG 
                 1507 
               
               
                   
                   
                 CACCCCAdTdT 
                   
                   
                 AAUGCUGdTdT 
                   
                 CACACCCCA 
                   
               
               
                   
               
               
                 AD-71208 
                 A-142795 
                 CACCCCAAGGGA 
                 1092 
                 A-142796 
                 UGGGUUGUCCCU 
                 1300 
                 CACCCCAAGG 
                 1508 
               
               
                   
                   
                 CAACCCAdTdT 
                   
                   
                 UGGGGUGdTdT 
                   
                 GACAACCCC 
                   
               
               
                   
               
               
                 AD-71209 
                 A-142797 
                 ACAACCCCAUCA 
                 1093 
                 A-142798 
                 UGUCAUAUGAUG 
                 1301 
                 ACAACCCCAU 
                 1509 
               
               
                   
                   
                 UAUGACAdTdT 
                   
                   
                 GGGUUGUdTdT 
                   
                 CAUAUGACA 
                   
               
               
                   
               
               
                 AD-71210 
                 A-142801 
                 ACCCUCUCCAUG 
                 1094 
                 A-142802 
                 UGUUGGGCAUGG 
                 1302 
                 ACCCUCUCCA 
                 1510 
               
               
                   
                   
                 CCCAACAdTdT 
                   
                   
                 AGAGGGUdTdT 
                   
                 UGCCCAACC 
                   
               
               
                   
               
               
                 AD-71211 
                 A-142803 
                 UGCCCAACCUAA 
                 1095 
                 A-142804 
                 UACAAUCUUAGG 
                 1303 
                 UGCCCAACCU 
                 1511 
               
               
                   
                   
                 GAUUGUAdTdT 
                   
                   
                 UUGGGCAdTdT 
                   
                 AAGAUUGUG 
                   
               
               
                   
               
               
                 AD-71212 
                 A-142805 
                 AAGAUUGUGUGG 
                 1096 
                 A-142806 
                 AAAAAACCCACA 
                 1304 
                 AAGAUUGUGU 
                 1512 
               
               
                   
                   
                 GUUUUUUdTdT 
                   
                   
                 CAAUCUUdTdT 
                   
                 GGGUUUUUU 
                   
               
               
                   
               
               
                 AD-71213 
                 A-142807 
                 UUUUUUAAUUAA 
                 1097 
                 A-142808 
                 AACAUUUUUAAU 
                 1305 
                 UUUUUUAAUU 
                 1513 
               
               
                   
                   
                 AAAUGUUdTdT 
                   
                   
                 UAAAAAAdTdT 
                   
                 AAAAAUGUU 
                   
               
               
                   
               
               
                 AD-71214 
                 A-142809 
                 UAAAAAUGUUAA 
                 1098 
                 A-142810 
                 AAAACUUUUAAC 
                 1306 
                 UAAAAAUGUU 
                 1514 
               
               
                   
                   
                 AAGUUUUdTdT 
                   
                   
                 AUUUUUAdTdT 
                   
                 AAAAGUUUU 
                   
               
               
                   
               
               
                 AD-71215 
                 A-142811 
                 AAAGUUUUAAAC 
                 1099 
                 A-142812 
                 UUUUCAUGUUUA 
                 1307 
                 AAAGUUUUAA 
                 1515 
               
               
                   
                   
                 AUGAAAAdTdT 
                   
                   
                 AAACUUUdTdT 
                   
                 ACAUGAAAA 
               
               
                   
               
            
           
         
       
     
     
       
         
           
               
             
               
                 TABLE 8 
               
             
            
               
                   
               
               
                 GCK Single Dose Screen in Primary Cynomolgus Hepatocytes 
               
            
           
           
               
               
               
               
            
               
                   
                 DuplexID 
                 20 nM_AVG 
                 20 nM_STDEV 
               
               
                   
                   
               
            
           
           
               
               
               
               
            
               
                   
                 AD-71009 
                 91.7 
                 8.2 
               
               
                   
                 AD-71010 
                 85.5 
                 11.8 
               
               
                   
                 AD-71011 
                 101.4 
                 19.6 
               
               
                   
                 AD-71012 
                 96.3 
                 20.8 
               
               
                   
                 AD-71013 
                 102.8 
                 21.7 
               
               
                   
                 AD-71014 
                 105.3 
                 47.3 
               
               
                   
                 AD-71015 
                 32.6 
                 6.6 
               
               
                   
                 AD-71016 
                 98.7 
                 19.1 
               
               
                   
                 AD-71017 
                 64.1 
                 16.4 
               
               
                   
                 AD-71018 
                 45.9 
                 12.8 
               
               
                   
                 AD-71019 
                 39.6 
                 8.5 
               
               
                   
                 AD-71020 
                 89.2 
                 23.9 
               
               
                   
                 AD-71021 
                 60.4 
                 7.7 
               
               
                   
                 AD-71022 
                 100.8 
                 12.7 
               
               
                   
                 AD-71023 
                 97.4 
                 33 
               
               
                   
                 AD-71024 
                 60.6 
                 18.8 
               
               
                   
                 AD-71025 
                 31.1 
                 9 
               
               
                   
                 AD-71026 
                 33.6 
                 10.5 
               
               
                   
                 AD-71027 
                 46.3 
                 14.6 
               
               
                   
                 AD-71028 
                 35.6 
                 9.6 
               
               
                   
                 AD-71029 
                 35.6 
                 15.9 
               
               
                   
                 AD-71030 
                 30.4 
                 7.9 
               
               
                   
                 AD-71031 
                 107.2 
                 29 
               
               
                   
                 AD-71032 
                 78.4 
                 15.7 
               
               
                   
                 AD-71033 
                 71.8 
                 13.9 
               
               
                   
                 AD-71034 
                 39.7 
                 14.3 
               
               
                   
                 AD-71035 
                 46.8 
                 11.4 
               
               
                   
                 AD-71036 
                 77.6 
                 21.3 
               
               
                   
                 AD-71037 
                 37.4 
                 15.2 
               
               
                   
                 AD-71038 
                 48.8 
                 19.6 
               
               
                   
                 AD-71039 
                 70.1 
                 9.5 
               
               
                   
                 AD-71040 
                 65.9 
                 16.2 
               
               
                   
                 AD-71041 
                 94.8 
                 18.6 
               
               
                   
                 AD-71042 
                 108.4 
                 24.2 
               
               
                   
                 AD-71043 
                 35.8 
                 12.2 
               
               
                   
                 AD-71044 
                 46.6 
                 10.9 
               
               
                   
                 AD-71045 
                 39.4 
                 9.5 
               
               
                   
                 AD-71046 
                 35.3 
                 3.9 
               
               
                   
                 AD-71047 
                 82 
                 24.4 
               
               
                   
                 AD-71048 
                 39.8 
                 10.1 
               
               
                   
                 AD-71049 
                 95.3 
                 6.8 
               
               
                   
                 AD-71050 
                 151.2 
                 22.4 
               
               
                   
                 AD-71051 
                 54 
                 13.5 
               
               
                   
                 AD-71052 
                 48.7 
                 8.1 
               
               
                   
                 AD-71053 
                 44 
                 10.7 
               
               
                   
                 AD-71054 
                 53.4 
                 8.8 
               
               
                   
                 AD-71055 
                 39.8 
                 9.4 
               
               
                   
                 AD-71056 
                 51.8 
                 34 
               
               
                   
                 AD-71057 
                 71 
                 7.1 
               
               
                   
                 AD-71058 
                 38.9 
                 3.6 
               
               
                   
                 AD-71059 
                 78.1 
                 17.8 
               
               
                   
                 AD-71060 
                 54 
                 14.4 
               
               
                   
                 AD-71061 
                 108.4 
                 27.2 
               
               
                   
                 AD-71062 
                 69.4 
                 6.7 
               
               
                   
                 AD-71063 
                 35.1 
                 11 
               
               
                   
                 AD-71064 
                 53.1 
                 13.8 
               
               
                   
                 AD-71065 
                 94 
                 11.6 
               
               
                   
                 AD-71066 
                 149.2 
                 13.3 
               
               
                   
                 AD-71067 
                 50.8 
                 15.2 
               
               
                   
                 AD-71068 
                 113.4 
                 23.7 
               
               
                   
                 AD-71069 
                 44.9 
                 6.4 
               
               
                   
                 AD-71070 
                 112.3 
                 24.3 
               
               
                   
                 AD-71071 
                 32.7 
                 5.3 
               
               
                   
                 AD-71072 
                 40.1 
                 10.2 
               
               
                   
                 AD-71073 
                 53 
                 12.5 
               
               
                   
                 AD-71074 
                 135.4 
                 25.3 
               
               
                   
                 AD-71075 
                 100.8 
                 31.5 
               
               
                   
                 AD-71076 
                 35.6 
                 7.3 
               
               
                   
                 AD-71077 
                 26.9 
                 4.7 
               
               
                   
                 AD-71078 
                 54.4 
                 11.8 
               
               
                   
                 AD-71079 
                 48 
                 6.2 
               
               
                   
                 AD-71080 
                 96.1 
                 6 
               
               
                   
                 AD-71081 
                 105.4 
                 7.9 
               
               
                   
                 AD-71082 
                 127 
                 20 
               
               
                   
                 AD-71083 
                 117.2 
                 33.2 
               
               
                   
                 AD-71084 
                 124.4 
                 20.7 
               
               
                   
                 AD-71085 
                 78.5 
                 6.6 
               
               
                   
                 AD-71086 
                 30.2 
                 12.7 
               
               
                   
                 AD-71087 
                 44.2 
                 2 
               
               
                   
                 AD-71088 
                 92.9 
                 21.9 
               
               
                   
                 AD-71089 
                 36.1 
                 13.7 
               
               
                   
                 AD-71090 
                 40 
                 3.6 
               
               
                   
                 AD-71091 
                 58 
                 3.3 
               
               
                   
                 AD-71092 
                 62.3 
                 14.9 
               
               
                   
                 AD-71093 
                 58.6 
                 18.2 
               
               
                   
                 AD-71094 
                 82.7 
                 22.9 
               
               
                   
                 AD-71095 
                 116.8 
                 27.6 
               
               
                   
                 AD-71096 
                 40.3 
                 11.8 
               
               
                   
                 AD-71097 
                 26.6 
                 6.9 
               
               
                   
                 AD-69448 
                 34.9 
                 9.3 
               
               
                   
                 AD-71098 
                 50.4 
                 7.4 
               
               
                   
                 AD-71099 
                 50.7 
                 18.4 
               
               
                   
                 AD-71100 
                 23.8 
                 1.7 
               
               
                   
                 AD-71101 
                 70.7 
                 18.1 
               
               
                   
                 AD-71102 
                 37.2 
                 5.5 
               
               
                   
                 AD-71103 
                 98.3 
                 17.6 
               
               
                   
                 AD-71104 
                 78.8 
                 22.8 
               
               
                   
                 AD-71105 
                 29.6 
                 6.1 
               
               
                   
                 AD-71106 
                 26.1 
                 8.1 
               
               
                   
                 AD-71107 
                 54.4 
                 9.2 
               
               
                   
                 AD-71108 
                 143.2 
                 28.5 
               
               
                   
                 AD-71109 
                 116.6 
                 15.9 
               
               
                   
                 AD-71110 
                 107.3 
                 21.5 
               
               
                   
                 AD-71111 
                 36.8 
                 9 
               
               
                   
                 AD-71112 
                 76.4 
                 19.7 
               
               
                   
                 AD-71113 
                 71.7 
                 12.9 
               
               
                   
                 AD-71114 
                 94.8 
                 22.5 
               
               
                   
                 AD-71115 
                 43.3 
                 12.3 
               
               
                   
                 AD-71116 
                 58.7 
                 7.9 
               
               
                   
                 AD-71117 
                 26.9 
                 7.1 
               
               
                   
                 AD-71118 
                 74 
                 15.8 
               
               
                   
                 AD-71119 
                 33.8 
                 12.9 
               
               
                   
                 AD-71120 
                 30.5 
                 4.3 
               
               
                   
                 AD-71121 
                 51.8 
                 7.6 
               
               
                   
                 AD-71122 
                 71.8 
                 20.4 
               
               
                   
                 AD-71123 
                 45.6 
                 12.4 
               
               
                   
                 AD-71124 
                 156.4 
                 6.6 
               
               
                   
                 AD-71125 
                 55.4 
                 6.2 
               
               
                   
                 AD-71126 
                 102.3 
                 8.5 
               
               
                   
                 AD-71127 
                 107.8 
                 18.2 
               
               
                   
                 AD-71128 
                 95.6 
                 19 
               
               
                   
                 AD-71129 
                 51.6 
                 16.7 
               
               
                   
                 AD-71130 
                 28.4 
                 10.3 
               
               
                   
                 AD-71131 
                 49 
                 6.5 
               
               
                   
                 AD-71132 
                 127.7 
                 25.6 
               
               
                   
                 AD-71133 
                 157.5 
                 17.4 
               
               
                   
                 AD-71134 
                 38 
                 11.1 
               
               
                   
                 AD-71135 
                 62.6 
                 7.6 
               
               
                   
                 AD-71136 
                 141.6 
                 20.3 
               
               
                   
                 AD-71137 
                 55.9 
                 6.4 
               
               
                   
                 AD-71138 
                 37.9 
                 5 
               
               
                   
                 AD-71139 
                 125.8 
                 27.6 
               
               
                   
                 AD-71140 
                 41.8 
                 2.5 
               
               
                   
                 AD-71141 
                 32.8 
                 6.7 
               
               
                   
                 AD-71142 
                 40.4 
                 11.5 
               
               
                   
                 AD-71143 
                 177.4 
                 27.3 
               
               
                   
                 AD-71144 
                 53.5 
                 9.1 
               
               
                   
                 AD-71145 
                 41.3 
                 27 
               
               
                   
                 AD-71146 
                 105.9 
                 13.9 
               
               
                   
                 AD-71147 
                 98.2 
                 27.5 
               
               
                   
                 AD-71148 
                 73.6 
                 8.9 
               
               
                   
                 AD-71149 
                 159 
                 24.3 
               
               
                   
                 AD-71150 
                 157.9 
                 31.1 
               
               
                   
                 AD-71151 
                 131.4 
                 4.3 
               
               
                   
                 AD-71152 
                 98.2 
                 25.2 
               
               
                   
                 AD-71153 
                 66.6 
                 23 
               
               
                   
                 AD-71154 
                 134.8 
                 11.9 
               
               
                   
                 AD-71155 
                 52.8 
                 4.2 
               
               
                   
                 AD-71156 
                 111.4 
                 43.9 
               
               
                   
                 AD-71157 
                 37.6 
                 7.7 
               
               
                   
                 AD-71158 
                 83.3 
                 16.9 
               
               
                   
                 AD-71159 
                 33.7 
                 7.3 
               
               
                   
                 AD-71160 
                 44.2 
                 7.2 
               
               
                   
                 AD-71161 
                 159.1 
                 28.3 
               
               
                   
                 AD-71162 
                 137.2 
                 20.9 
               
               
                   
                 AD-71163 
                 23.4 
                 5.4 
               
               
                   
                 AD-71164 
                 27.7 
                 2.6 
               
               
                   
                 AD-71165 
                 38.1 
                 8 
               
               
                   
                 AD-71166 
                 46.4 
                 11.4 
               
               
                   
                 AD-71167 
                 53.1 
                 17.6 
               
               
                   
                 AD-71168 
                 130.3 
                 14 
               
               
                   
                 AD-71169 
                 95.8 
                 24.4 
               
               
                   
                 AD-71170 
                 108.8 
                 15.4 
               
               
                   
                 AD-71171 
                 57.6 
                 5.3 
               
               
                   
                 AD-71172 
                 75 
                 26.8 
               
               
                   
                 AD-71173 
                 87.3 
                 24 
               
               
                   
                 AD-71174 
                 62.7 
                 24.2 
               
               
                   
                 AD-71175 
                 83.6 
                 25.8 
               
               
                   
                 AD-71176 
                 26.7 
                 3.5 
               
               
                   
                 AD-71177 
                 31 
                 4.7 
               
               
                   
                 AD-71178 
                 114.6 
                 20 
               
               
                   
                 AD-71179 
                 101.9 
                 25.9 
               
               
                   
                 AD-71180 
                 114 
                 12.4 
               
               
                   
                 AD-71181 
                 38.2 
                 10.1 
               
               
                   
                 AD-71182 
                 32.3 
                 3.2 
               
               
                   
                 AD-71183 
                 49.9 
                 7.4 
               
               
                   
                 AD-71184 
                 60.6 
                 6.7 
               
               
                   
                 AD-71185 
                 30 
                 3 
               
               
                   
                 AD-71186 
                 55.7 
                 16.3 
               
               
                   
                 AD-71187 
                 72.8 
                 15.2 
               
               
                   
                 AD-71188 
                 32.1 
                 2.2 
               
               
                   
                 AD-71189 
                 76.4 
                 13.9 
               
               
                   
                 AD-71190 
                 34.2 
                 9.1 
               
               
                   
                 AD-71191 
                 51.7 
                 9.5 
               
               
                   
                 AD-71192 
                 87.5 
                 8.3 
               
               
                   
                 AD-71193 
                 110.2 
                 20.9 
               
               
                   
                 AD-71194 
                 56.9 
                 12.6 
               
               
                   
                 AD-71195 
                 63.1 
                 25.2 
               
               
                   
                 AD-71196 
                 38.1 
                 10.3 
               
               
                   
                 AD-71197 
                 34 
                 9.8 
               
               
                   
                 AD-71198 
                 108.1 
                 12 
               
               
                   
                 AD-71199 
                 117.9 
                 11.5 
               
               
                   
                 AD-71200 
                 50.1 
                 12.3 
               
               
                   
                 AD-71201 
                 38.2 
                 5.8 
               
               
                   
                 AD-71202 
                 73.9 
                 3.6 
               
               
                   
                 AD-71203 
                 110.3 
                 9.2 
               
               
                   
                 AD-71204 
                 140 
                 11 
               
               
                   
                 AD-71205 
                 35.8 
                 4.5 
               
               
                   
                 AD-71206 
                 28 
                 12.1 
               
               
                   
                 AD-71207 
                 22.4 
                 11 
               
               
                   
                 AD-71208 
                 54.6 
                 10.9 
               
               
                   
                 AD-71209 
                 40.6 
                 14.9 
               
               
                   
                 AD-71210 
                 41.5 
                 6.3 
               
               
                   
                 AD-71211 
                 54.4 
                 16 
               
               
                   
                 AD-71212 
                 122.4 
                 20.7 
               
               
                   
                 AD-71213 
                 111.4 
                 13.7 
               
               
                   
                 AD-71214 
                 120.1 
                 19.5 
               
               
                   
                 AD-71215 
                 98 
                 11.7