Compositions and methods for regulating PNPLA3 expression

Abstract

Disclosed are oligonucleotides and compositions comprising the same for inhibiting or reducing patatin-like phospholipase domain-containing protein 3 (PNPLA3) gene expression. Methods for making and using the oligonucleotides are also disclosed, particularly for use in treating diseases, disorders, and / or conditions associated with PNPLA3 expression.

Description

Technical Field

[0001] Sequence Listing This application includes a sequence listing submitted electronically in ASCII format, which is hereby incorporated by reference in its entirety. The above ASCII copy, created on April 11, 2022, is named 400930-028WO_190156_SL.txt and is 374,357 bytes in size.

[0002] (Cross - reference to Related Applications) This application claims the benefit of priority under 35 U.S.C. § 119(e) to U.S. Provisional Application No. 63 / 174,932, filed on April 14, 2021, which is hereby incorporated by reference in its entirety.

[0003] This disclosure generally relates to biology and medicine, and more particularly, to the use of oligonucleotide compositions for inhibiting or reducing patatin - like phospholipase domain - containing protein 3 (PNPLA3) gene expression, and to the use thereof, particularly for the treatment of diseases, disorders, and / or conditions associated with PNPLA3.

Background Art

[0004] PNPLA3 is a protein encoded by the PNPLA3 gene and has triacylglycerol lipase activity and acyl - glycerol transacylase activity. Human PNPLA3 is highly expressed in the liver and moderately expressed in adipose tissue, brain, kidney, and skin.

[0005] This specification specifically focuses on the mutation at position 148 of human PNPLA3 from isoleucine (Ile / I) to methionine (Met / M) (I148M or PNPLA3 148M; i.e., PNPLA3 rs738409). See Pingitore & Romeo (2019) Biochim. Biophys. Acta Mol. Cell Biol. Lipids 1864:900-906. Compared to wild-type PNPLA3, PNPLA3 148M is thought to lack lipase activity but have increased transacylase activity. See Kumari et al. (2012) Cell Metab. 15:691-702.

[0006] PNPLA3 148M is strongly associated with a wide range of liver diseases resulting from triglyceride (TG) accumulation, liver damage, and fibrosis, including alcoholic hepatitis (AH), alcoholic liver disease (ALD), cirrhosis, hepatocellular carcinoma (HCC), non-alcoholic fatty liver disease (NAFLD), and non-alcoholic steatohepatitis (NASH). It is thought that the degradation of PNPLA3 148M by the proteasome is delayed, resulting in protein accumulation on the surface of lipid droplets, which prevents other proteins from metabolizing TG in hepatocytes. Therefore, reducing the expression of PNPLA3 148M may allow other lipases to function normally and potentially reverse the adverse effects of PNPLA3 148M across various pathological conditions.

[0007] Several RNA-based therapeutics are known to attempt to inhibit or reduce PNPLA3 expression. For example, International Patent Application Publications WO2016 / 130806 and WO2019 / 118638 describe double-stranded (ds)RNAi constructs for inhibiting or reducing PNPLA3 expression, and methods of using them to treat or prevent liver diseases such as NAFLD. Also, International Patent Application Publication WO2020 / 061200 describes antisense oligonucleotides for inhibiting or reducing PNPLA3 expression.

[0008] Despite the existence of several PNPLA3-targeting therapies, there is a need for further treatments that inhibit or reduce PNPLA3 expression to treat liver disease. [Overview of the Initiative]

[0009] To address this need, this disclosure describes compositions and methods for treating diseases, disorders, and / or conditions associated with PNPLA3 expression. This disclosure is partly based on the discovery and development of ds oligonucleotides (e.g., RNAi oligonucleotides) for selectively inhibiting and / or reducing PNPLA3 expression, for example, in the liver. Thus, target sequences within PNPLA3 were identified, and RNAi oligonucleotides were generated that bind to these target sequences to inhibit PNPLA3 mRNA expression. As shown herein, RNAi oligonucleotides inhibit human and cynomolgus monkey PNPLA3 expression in the liver. While not limited to theory, the RNAi oligonucleotides herein are useful for treating diseases, disorders, or conditions associated with PNPLA3 expression (e.g., liver diseases, e.g., AH, ALD, cirrhosis, HCC, cholangiocarcinoma (CCA), primary sclerosing cholangitis (PSC), NAFLD, and NASH). In general, the RNAi oligonucleotides described herein are useful for treating diseases, disorders, or conditions associated with abnormal PNPLA3 expression (e.g., mutant PNPLA3 allele expression). In particular, the RNAi oligonucleotides described herein are useful for treating diseases, disorders, or conditions associated with mutant PNPLA3 expression.

[0010] Therefore, this disclosure refers to Table 1 (for example, Sequence IDs 9, 11, 13, 15, 17, 19, 21, 23, 25, 27, 29, 31, 33, 35, 37, 39, 41, 43, 45, 47, 49, 51, 53, 55, 57, 59, 61, 63, 65, 67, 69, 71, 73, 75, 77, 79, 81, 83, 85, 87, 89, 91, 93, 95, 97, 99, 101, 103, 105, 107, 109, 111, 113, 115, 117, 119, 121, 123, 125, 127, 129, 131, 133, 135, 137, 139, 141, 143, 14 5, 147, 149, 151, 153, 155, 157, 159, 161, 163, 165, 167, 169, 171, 173, 175, 177, 179, 181, 183, 185, 187, 189, 191, 193, 195, 197, 199, 201, 203, 205, 207 ,209,211,213,215,217,219,221,223,225,227,229,231,233,235,237,239,241,243,245,247,249,251,253,255,257,259,261,263,265,267,269, 271, 273, 275, 277, 279, 281, 283, 285, 287, 289, 291, 293, 295, 297, 299, 301, 303, 305, 307, 309, 311, 313, 315, 317, 319, 321, 323, 325, 327, 329, 331, 3 33, 335, 337, 339, 341, 343, 345, 347, 349, 351, 353, 355, 357, 359, 361, 363, 365, 367, 369, 371, 373, 375, 377, 379, 381, 383, 385, 387, 389, 391, 393, 39 5, 397, 399, 401, 403, 405, 407, 409, 411, 413, 415, 417, 419, 421, 423, 425, 427, 429, 431, 433, 435, 437, 439, 441, 443, 445, 447, 449, 451, 453, 455, 457 ,459,461,463,465,467,469,471,473,475,477,479,481,483,485,487,489,491,493,495,497,499,501,503,505,507,509,511,513,515,517,519,521, 523, 525, 527, 529, 531, 533, 535, 537, 539, 541, 543, 545, 547, 549, 551, 553, 555, 557, 559, 561, 563, 565, 567, 569, 571, 573, 575, 577, 579, 581, 5 83, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, 629, 631, 633, 635, 637, 639, 641, 643, 64 5, 647, 649, 651, 653, 655, 657, 659, 661, 663, 665, 667, 669, 671, 673, 675, 677, 679, 681, 683, 685, 687, 689, 691, 693, 695, 697, 699, 701, 703, 705, 707 ,709,711,713,715,717,719,721,723,725,727,729,731,733,735,737,739,741,743,745,747,749,751,753,755,757,759,761,763,765,767,769, 771, 773, and 775) or Table 3 (for example, Sequence IDs 777, 779, 781, 783, 785, 787, 789, 791, 793, 795, 797, 799, 801, 803, 805, 807, 809, 811, 813, 815, 817, 819, 821, 823, 825, 827, 829, 831, 833, 835, 837, 839, 841, 843, 845, 847, 849, 851, 853, 855, 857, 859, 861, 863, 865, 867, 869, 871, 873, 875, 877, 879, 881, 883, 885, 88 7, 889, 891, 893, 895, 897, 899, 901, 903, 905, 907, 909, 911, 913, 915, 917, 919, 921, 923, 925, 927, 929, 931, 933, 935, 937, 939, 941, 943, 945, 947, 949 ,951,953,955,957,959,961,963,965,967,969,971,973,975,977,979,981,983,985,987,989,991,993,995,997,999,1001,1003,1005,1007,1009,1011, 1013, 1015, 1017, 1019, 1021, 1023, 1025, 1027, 1029, 1031, 1033, 1035, 1037, 1039, 1041, 1043, 1045, 1047, 1049, 1051, 1053, 1055, 1057, 1059, 10 61, 1063, 1065, 1067, 1069, 1071, 1073, 1075, 1077, 1079, 1081, 1083, 1085, 1087, 1089, 1091, 1093, 1095, 1097, 1099, 1101, 1103, 1105, 1107, 1109, 1111, This document describes RNAi oligonucleotides for reducing or inhibiting PNPLA3 expression, which include a sense strand having one of the sequences described in 1113, 1115, 1117, 1119, 1121, 1123, 1125, 1127, 1129, 1131, 1133, 1135, 1137, 1139, 1141, 1143, 1145, 1147, 1149, 1151, 1153, 1155, 1157, 1159, 1161, and 1163, particularly sequence numbers 787, 843, 867, 871, 937, 1003, 1007, 1017, 1161, or 1163.

[0011] Alternatively, this disclosure refers to Table 1 (for example, Sequence IDs 10, 12, 14, 16, 18, 20, 22, 24, 26, 28, 30, 32, 34, 36, 38, 40, 42, 44, 46, 48, 50, 52, 54, 56, 58, 60, 62, 64, 66, 68, 70, 72, 74, 76, 78, 80, 82, 84, 86, 88, 90, 92, 94, 96, 98, 100, 102, 104, 106, 108, 110, 112, 114, 116, 118, 120, 122, 124, 126, 128, 130, 132, 134, 136, 138, 140, 142, 144, 14 6, 148, 150, 152, 154, 156, 158, 160, 162, 164, 166, 168, 170, 172, 174, 176, 178, 180, 182, 184, 186, 188, 190, 192, 194, 196, 198, 200, 202, 204, 206, 208 ,210,212,214,216,218,220,222,224,226,228,230,232,234,236,238,240,242,244,246,248,250,252,254,256,258,260,262,264,266,268,270, 272, 274, 276, 278, 280, 282, 284, 286, 288, 290, 292, 294, 296, 298, 300, 302, 304, 306, 308, 310, 312, 314, 316, 318, 320, 322, 324, 326, 328, 330, 332, 3 34, 336, 338, 340, 342, 344, 346, 348, 350, 352, 354, 356, 358, 360, 362, 364, 366, 368, 370, 372, 374, 376, 378, 380, 382, ​​384, 386, 388, 390, 392, 394, 39 6, 398, 400, 402, 404, 406, 408, 410, 412, 414, 416, 418, 420, 422, 424, 426, 428, 430, 432, 434, 436, 438, 440, 442, 444, 446, 448, 450, 452, 454, 456, 458 ,460,462,464,466,468,470,472,474,476,478,480,482,484,486,488,490,492,494,496,498,500,502,504,506,508,510,512,514,516,518,520,522, 524, 526, 528, 530, 532, 534, 536, 538, 540, 542, 544, 546, 548, 550, 552, 554, 556, 558, 560, 562, 564, 566, 568, 570, 572, 574, 576, 578, 580, 582, 584, 586, 588, 590, 592, 594, 596, 598, 600, 602, 604, 606, 608, 610, 612, 614, 616, 618, 620, 622, 624, 626, 628, 630, 632, 634, 636, 638, 640, 642, 644, 6 46, 648, 650, 652, 654, 656, 658, 660, 662, 664, 666, 668, 670, 672, 674, 676, 678, 680, 682, 684, 686, 688, 690, 692, 694, 696, 698, 700, 702, 704, 706, 7 08, 710, 712, 714, 716, 718, 720, 722, 724, 726, 728, 730, 732, 734, 736, 738, 740, 742, 744, 746, 748, 750, 752, 754, 756, 758, 760, 762, 764, 766, 768, 77 (0, 772, 774, and 776) or Table 3 (for example, Sequence IDs 778, 780, 782, 784, 786, 788, 790, 792, 794, 796, 798, 800, 802, 804, 806, 808, 810, 812, 814, 816, 818, 820, 822, 824, 826, 828, 830, 832, 834, 836, 838, 840, 842, 844, 846, 848, 850, 852, 854, 856, 858, 860, 862, 864, 866, 868, 870, 872, 874, 876, 878, 880, 882, 884, 886) ,888,890,892,894,896,898,900,902,904,906,908,910,912,914,916,918,920,922,924,926,928,930,932,934,936,938,940,942,944,946,948, 950, 952, 954, 956, 958, 960, 962, 964, 966, 968, 970, 972, 974, 976, 978, 980, 982, 984, 986, 988, 990, 992, 994, 996, 998, 1000, 1002, 1004, 1006, 1008,1010, 1012, 1014, 1016, 1018, 1020, 1022, 1024, 1026, 1028, 1030, 1032, 1034, 1036, 1038, 1040, 1042, 1044, 1046, 1048, 1050, 1052, 1054, 1056, 1058, 1060 ,1062,1064,1066,1068,1070,1072,1074,1076,1078,1080,1082,1084,1086,1088,1090,1092,1094,1096,1098,1100,1102,1104,1106,1108,1110,1112 This document describes RNAi oligonucleotides for reducing or inhibiting PNPLA3 expression, which include an antisense strand having one of the sequences described in 1114, 1116, 1118, 1120, 1122, 1124, 1126, 1128, 1130, 1132, 1134, 1136, 1138, 1140, 1142, 1144, 1146, 1148, 1150, 1152, 1154, 1156, 1158, 1160, 1162, and 1164, particularly sequence numbers 788, 844, 868, 872, 938, 1004, 1008, 1018, 1162, or 1164.

[0012] In certain embodiments, this disclosure relates to sequences listed in Table A, B, C, or D (for example, sequence numbers 1178, 1180, 1182, 1184, 1186, 1188, 1190, 1192, 1194, 1196, 1198, 1200, 1202, 1204, 1206, 1208, 1210, 1212, 1214, 1216, 1218, 1220, 1222, 1224, 1226, 1228, 1230, 1232, 1234, 1236, 1238, 1240, 124 RNAi oligonucleotides for reducing or inhibiting PNPLA3 expression include sense strands having 2, 1244, 1246, 1248, 1250, 1252, 1254, 1256, 1258, 1260, 1262, 1264, 1266, 1268, 1270, 1272, 1274, 1276, 1278, 1280, 1282, 1284, 1286, 1288, 1290, 1292, 1294, 1296, 1298, or 1300).

[0013] In certain embodiments, this disclosure relates to the sequences listed in Table A, B, C, or D (for example, sequence numbers 1179, 1181, 1183, 1185, 1187, 1189, 1191, 1193, 1195, 1197, 1199, 1201, 1203, 1205, 1207, 1209, 1211, 1213, 1215, 1217, 1219, 1221, 1223, 1225, 1227, 1229, 1231, 1233, 1235, 1237, 1239, 1241, 1243 This document describes RNAi oligonucleotides for reducing or inhibiting PNPLA3 expression, which include antisense strands having 1245, 1247, 1249, 1251, 1253, 1255, 1257, 1259, 1261, 1263, 1265, 1267, 1269, 1271, 1273, 1275, 1277, 1279, 1281, 1283, 1285, 1287, 1289, 1291, 1293, 1295, 1297, 1299, or 1301).

[0014] In some embodiments, RNAi oligonucleotides for reducing or inhibiting PNPLA3 expression, comprising an antisense strand and a sense strand, wherein the antisense strand has a sequence described in Table 1 or Table 3, and the sense strand has a sequence described in Table 1 or Table 3, are described.

[0015] In some embodiments, RNAi oligonucleotides for reducing or inhibiting PNPLA3 expression, comprising an antisense strand and a sense strand, wherein the antisense strand has a sequence described in Table A, Table B, Table C, or Table D, and the sense strand has a sequence described in Table A, Table B, Table C, or Table D, are described.

[0016] In some embodiments, RNAi oligonucleotides for reducing or inhibiting PNPLA3 expression include an antisense strand and a sense strand, wherein the antisense strand and the sense strand form a double-stranded region, and the antisense strand has a complementary region to one of the PNPLA3 mRNA target sequences of sequence numbers 1167-1176.

[0017] In any of the embodiments described above, the antisense chain is approximately 15 to 30 nucleotides long. In some embodiments, the antisense chain is approximately 20 to 25 nucleotides long. In some embodiments, the antisense chain is 22 nucleotides long.

[0018] In any of the embodiments described above, the sense strand is approximately 15 to 50 nucleotides long. In some examples, the sense strand is approximately 20 to 40 nucleotides long. In some embodiments, the sense strand is 36 nucleotides long.

[0019] In any of the embodiments described above, the double-stranded region is approximately 19 to 21 nucleotides long. In a particular embodiment, the double-stranded region is 20 nucleotides long.

[0020] In any of the embodiments described above, the complementary region is at least 15 consecutive nucleotides long. In some embodiments, the complementary region is about 19 to about 21 consecutive nucleotides long. In other embodiments, the complementary region is 19 or 21 consecutive nucleotides long.

[0021] In any of the embodiments described above, the RNAi oligonucleotide includes a stem-loop at the 3' end of the sense strand, represented as S1-L-S2 (where S1 is complementary to S2, and L forms a loop between S1 and S2 approximately 3 to 5 nucleotides long).

[0022] In any of the above embodiments, the antisense strand, the sense strand, or both have an overhang array. In some embodiments, the antisense strand includes a 3' overhang that is at least 1 nucleotide in length. In other embodiments, the 3' overhang array is 2 nucleotides in length, for example GG.

[0023] An oligonucleotide comprising an antisense strand and a sense strand, wherein the antisense strand can be about 21 nucleotides to about 27 nucleotides in length, has a complementary region to PNPLA3, the sense strand has a stem-loop shown as S1-L-S2 at its 3' end (S1 is complementary to S2, and L forms a loop about 3 nucleotides to about 5 nucleotides in length between S1 and S2), and the antisense strand and the sense strand form a double-stranded structure that is at least about 19 nucleotides in length but not covalently linked, is also described.

[0024] In some embodiments, loop L is a triloop or a tetraloop. In some examples, L is a 4-nucleotide tetraloop. In other embodiments, L includes the sequence 5'-GAAA-3'.

[0025] In some embodiments, S1 and S2 are 1 to 10 nucleotides in length and have the same length. In other embodiments, S1 and S2 are 1 nucleotide in length, 2 nucleotides in length, 3 nucleotides in length, 4 nucleotides in length, 5 nucleotides in length, 6 nucleotides in length, 7 nucleotides in length, 8 nucleotides in length, 9 nucleotides in length, or 10 nucleotides in length. In other embodiments, S1 and S2 are 6 nucleotides in length. In certain embodiments, the stem-loop includes the sequence 5'-GCAGCCGAAAGGCUGC-3' (SEQ ID NO: 1177).

[0026] In some embodiments, the antisense strand is 27 nucleotides in length and the sense strand is 25 nucleotides in length. In other embodiments, the antisense strand is 22 nucleotides in length and the sense strand is 36 nucleotides in length.

[0027] In the above embodiment, the double-stranded region includes a 3'-overhang sequence on the antisense strand. In some embodiments, the 3'-overhang sequence on the antisense strand is 2 nucleotides in length.

[0028] In any of the above embodiments, at least one nucleotide in the oligonucleotide is a modified nucleotide. In some examples, modified nucleotides include 2'-modifications such as 2'-aminoethyl, 2'-fluoro, 2'-O-methyl, 2'-O-methoxyethyl, and 2'-deoxy-2'-fluoro-β-arabinonucleic acid. In certain examples, all nucleotides in the oligonucleotide include a 2'-modification such as 2'-fluoro or 2'-O-methyl.

[0029] In any of the above embodiments, at least one nucleotide in the oligonucleotide includes a modified internucleotide linkage. In some embodiments, the modified internucleotide linkage is a phosphorothioate linkage.

[0030] In any of the above embodiments, the 4'-carbon of the sugar of the 5'-nucleotide of the antisense strand includes a phosphate analog such as oxymethylphosphonate, vinylphosphonate, or malonylphosphonate. Alternatively, or optionally, the phosphate analog is a 4'-phosphate analog such as 5'-methoxyphosphonate-4'-oxy.

[0031] In any of the above embodiments, at least one nucleotide of the oligonucleotide can be conjugated to one or more targeting ligands such as amino sugars, carbohydrates, cholesterol, lipids, or polypeptides. In some embodiments, the targeting ligand is an N-acetylgalactosamine (GalNAc) moiety. In other embodiments, the GalNAc moiety is a monovalent GalNAc moiety, a divalent GalNAc moiety, a trivalent GalNAc moiety, or a tetravalent GalNAc moiety.

[0032] In some embodiments, the targeting ligand is conjugated to one or more nucleotides of the stem-loop L. In specific examples, up to four nucleotides of the stem-loop L are each conjugated to a monovalent GalNAc portion.

[0033] In any of the embodiments described above, the oligonucleotide is an RNAi oligonucleotide. In some examples, the RNAi oligonucleotide comprises a sense strand having one of the nucleotide sequences listed in Table 1 or Table 3, in particular SEQ ID NOs: 787, 843, 867, 871, 937, 1003, 1007, 1017, 1161, or 1163. In certain embodiments, the RNAi oligonucleotide comprises a sense strand having one of the nucleotide sequences listed in Tables A, B, C, or D, in particular SEQ ID NOs: 1188, 1190, 1220, 1224, 1230, 1232, 1244, 1246, 1250, or 1254. In some examples, the RNAi oligonucleotide comprises an antisense strand having one of the nucleotide sequences listed in Table 1 or Table 3, in particular SEQ ID NOs: 788, 844, 868, 872, 938, 1004, 1008, 1018, 1162, or 1164. In some embodiments, the RNAi oligonucleotide comprises an antisense strand having one of the nucleotide sequences listed in Tables A, B, C, or D, particularly SEQ ID NOs: 1189, 1191, 1221, 1225, 1231, 1233, 1245, 1247, 1251, or 1255. In certain examples, an RNAi oligonucleotide comprises a sense strand having one of the nucleotide sequences of SEQ ID NOs: 787, 843, 867, 871, 937, 1003, 1007, 1017, 1161, or 1163, or one of the nucleotide sequences of SEQ ID NOs: 1188, 1190, 1220, 1224, 1230, 1232, 1244, 1246, 1250, or 1254, and an antisense strand having one of the nucleotide sequences of SEQ ID NOs: 788, 844, 868, 872, 938, 1004, 1008, 1018, 1162, or 1164, or one of the nucleotide sequences of SEQ ID NOs: 1189, 1191, 1221, 1225, 1231, 1233, 1245, 1247, 1251, or 1255. In certain embodiments, the sense strand and antisense strand of the RNAi oligonucleotide are selected from the following, respectively: (a) Sequence IDs 787 and 788, (b) Sequence IDs 843 and 844, (c) Sequence IDs 867 and 868, (d) Sequence IDs 871 and 872, (e) Sequence IDs 937 and 938, (f) Sequence IDs 1003 and 1004, (g) Sequence IDs 1007 and 1008, (h) Sequence IDs 1017 and 1018, (i) Sequence IDs 1161 and 1162, and (j) Sequence IDs 1163 and 1164.

[0034] In some examples, RNAi oligonucleotides are nucleotide sequences listed in Table A, Table B, Table C, or Table D, particularly SEQ ID NOs: 1178, 1180, 1182, 1184, 1186, 1188, 1190, 1192, 1194, 1196, 1198, 1200, 1202, 1204, 1206, 1208, 1210, 1212, 1214, 1216, 1218, 1220, 1222, 1224, 1226, 1228, 1230, 12 Includes a sense chain having one of the following: 32, 1234, 1236, 1238, 1240, 1242, 1244, 1246, 1248, 1250, 1252, 1254, 1256, 1258, 1260, 1262, 1264, 1266, 1268, 1270, 1272, 1274, 1276, 1278, 1280, 1282, 1284, 1286, 1288, 1290, 1292, 1294, 1296, 1298, or 1300. In some examples, RNAi oligonucleotides are nucleotide sequences listed in Tables A, B, C, or D, particularly SEQ ID NOs: 1179, 1181, 1183, 1185, 1187, 1189, 1191, 1193, 1195, 1197, 1199, 1202, 1203, 1205, 1207, 1209, 1211, 1213, 1215, 1217, 1219, 1221, 1223, 1225, 1227, 1229, 1231, 123 The antisense chain includes one of the following: 3, 1235, 1237, 1239, 1241, 1243, 1245, 1247, 1249, 1251, 1253, 1255, 1257, 1259, 1261, 1263, 1265, 1267, 1269, 1271, 1273, 1275, 1277, 1279, 1281, 1283, 1285, 1287, 1289, 1291, 1293, 1295, 1297, 1299, or 1301.In a specific example, RNAi oligonucleotides are sequence numbers 1178, 1180, 1182, 1184, 1186, 1188, 1190, 1192, 1194, 1196, 1198, 1200, 1202, 1204, 1206, 1208, 1210, 1212, 1214, 1216, 1218, 1220, 1222, 1224, 1226, 1228, 1230, 1232, 1234, 1236, 12 It has one of the following nucleotide sequences: 38, 1240, 1242, 1244, 1246, 1248, 1250, 1252, 1254, 1256, 1258, 1260, 1262, 1264, 1266, 1268, 1270, 1272, 1274, 1276, 1278, 1280, 1282, 1284, 1286, 1288, 1290, 1292, 1294, 1296, 1298, or 1300. The sense chain and sequence numbers 1179, 1181, 1183, 1185, 1187, 1189, 1191, 1193, 1195, 1197, 1199, 1201, 1203, 1205, 1207, 1209, 1211, 1213, 1215, 1217, 1219, 1221, 1223, 1225, 1227, 1229, 1231, 1233, 1235, 1237, 1239, 1241, 1243, 1 It comprises an antisense strand having one of the following nucleotide sequences: 245, 1247, 1249, 1251, 1253, 1255, 1257, 1259, 1261, 1263, 1265, 1267, 1269, 1271, 1273, 1275, 1277, 1279, 1281, 1283, 1285, 1287, 1289, 1291, 1293, 1295, 1297, 1299, or 1301.

[0035] In a specific example, the RNAi oligonucleotide is one of the following nucleotide sequences: 1188, 1190, 1200, 1216, 1218, 1220, 1224, 1230, 1232, 1234, 1244, 1246, 1250, 1254, 1262, 1288, 1290, 1292, 1294, 1296, 1298, or 1300. It comprises a sense strand having a column and an antisense strand having one of the following nucleotide sequences: SEQ ID NOs: 1189, 1191, 1201, 1215, 1217, 1219, 1225, 1231, 1233, 1235, 1245, 1247, 1251, 1255, 1263, 1289, 1291, 1295, 1297, 1299, or 1301.

[0036] In a specific example, the sense and antisense strands of the RNAi oligonucleotide are selected from the following: (a) Sequence IDs 1220 and 1221, (b) Sequence IDs 1224 and 1225, (c) Sequence IDs 1230 and 1231, (d) Sequence IDs 1232 and 1233, (e) Sequence IDs 1188 and 1189, (f) Sequence IDs 1190 and 1191, (g) Sequence IDs 1244 and 1245, (h) Sequence IDs 1250 and 1251-, (i) Sequence IDs 1254 and 1255, and (j) Sequence IDs 1246 and 1247.

[0037] Also described is an oligonucleotide for inhibiting or reducing PNPLA3 expression, comprising a sense strand and an antisense strand, wherein the sense strand and the antisense strand form a double-stranded region, all nucleotides of the sense strand and the antisense strand include modifications of bases, sugars, and / or internucleotide bonds, and the antisense strand includes a complementary region to one of the PNPLA3 mRNA target sequences from SEQ ID NOs. 1167-1176, wherein the complementary region is at least about 15 consecutive nucleotides long.

[0038] In other embodiments, pharmaceutical compositions comprising at least one oligonucleotide of this specification and a pharmaceutically acceptable carrier, delivery agent, or excipient are described. In some examples, the pharmaceutical composition includes an additional therapeutic agent, such as an antidiabetic agent or an anti-obesity agent.

[0039] In other embodiments, methods for reducing PNPLA3 expression in cells, cell populations, tissues, organs, or organisms are described, comprising at least one step of administering / contacting the cells, cell populations, tissues, organs, or organisms with the oligonucleotides or pharmaceutical compositions of the Specified herein. In some examples, the reduction of PNPLA3 expression includes reducing the amount or level of PNPLA3 mRNA, the amount or level of PNPLA3 protein, or both, in cells, cell populations, tissues, organs, or organisms. In some examples, the cells, cell populations, tissues, organs, or organisms have a disease, disorder, or condition associated with PNPLA3 expression. In specific examples, the diseases, disorders, or conditions associated with PNPLA3 expression are cardiovascular metabolic diseases, AH, ALD, cirrhosis, HCC, CCA, and other cholangiopathies (such as PSC), NAFLD, and NASH.

[0040] In other embodiments, methods for treating individuals having or suspected to have a disease, disorder, or condition associated with PNPLA3 expression are described. These methods include at least one step of administering an effective amount of the oligonucleotide or pharmaceutical composition of this specification to an individual in need. In some examples, diseases, disorders, or conditions associated with PNPLA3 expression include cardiovascular metabolic disorders, AH, ALD, CCA, cirrhosis, HCC, NAFLD, PSC, and NASH. In some examples, the oligonucleotide or pharmaceutical composition is administered subcutaneously daily, weekly, monthly, quarterly, annually, or in particular monthly or quarterly.

[0041] In some cases, individuals have cirrhosis, diabetes, hepatic fibrosis, hepatitis, hyperlipidemia, AH, ALD, CCA, cirrhosis, HCC, NAFLD, PSC, NASH, obesity, and / or steatosis.

[0042] In any of the embodiments described above, the method includes additional steps, such as measuring or obtaining genotype information, PNPLA3 expression, PNPLA3 protein levels, body weight and / or blood glucose and / or TG of an individual, and evaluating the effectiveness of contact or administration by comparing such obtained values ​​with one or more baseline values ​​or previously obtained values. In some embodiments, the additional step includes confirming that the individual has the PNPLA3 I148M variant. In some embodiments, the additional step includes confirming that the individual does not have the PNPLA3 E434K variant. In some embodiments, the additional step includes confirming that the individual does not have the HSD17B13 variant (rs72613567) that cleaves the protein.

[0043] In any of the embodiments described above, the method may include administering the RNAi oligonucleotide or pharmaceutical composition simultaneously, separately, or sequentially with the second composition or second therapeutic agent. In some embodiments, the second composition or second therapeutic agent is a PNPLA3 antibody or a fragment thereof, an antidiabetic agent, or an anti-obesity agent. In some embodiments, the second composition or second therapeutic agent is administered at the same frequency as the RNAi oligonucleotide (i.e., every other day, twice a week, or even weekly). In other embodiments, the second composition or second therapeutic agent is administered at a different frequency than the RNAi oligonucleotide. Similarly, in other embodiments, the second composition or second therapeutic agent is administered via the same route as the RNAi oligonucleotide (e.g., subcutaneously). In yet another embodiment, the second composition or second therapeutic agent is administered via a different route than the RNAi oligonucleotide.

[0044] In other embodiments, the use of RNAi oligonucleotides of this specification for treating diseases, disorders, or conditions associated with PNPLA3 expression is described, wherein the RNAi oligonucleotides are administered separately, concurrently, or sequentially (i.e., in combination) with, optionally, a second composition or a second therapeutic agent.

[0045] In other embodiments, the use of RNAi oligonucleotides of this specification in the manufacture of a medicament for treating a disease, disorder, or condition related to PNPLA3 expression, the medicament further optionally comprising a second composition or a second therapeutic agent.

[0046] In other embodiments, a kit is described comprising at least one oligonucleotide of this specification, an optional pharmaceutically acceptable carrier, and a package insert having instructions for administering them to an individual having a disease, disorder, or condition related to PNPLA3 expression.

[0047] The advantage of the oligonucleotides and compositions described herein is that the suppression of PNPLA3 expression, particularly PNPLA3 148M, exerts beneficial effects on NAFLD in all areas, including fibrosis.

[0048] Other advantages, effects, features, and objectives will become more readily apparent upon consideration of the following embodiments for carrying out the invention. Such embodiments can be found in the following drawings. [Brief explanation of the drawing]

[0049] [Figure 1] A schematic diagram showing the structure and chemical modification pattern of a typical GalNAc-conjugated PNPLA3 oligonucleotide is disclosed. Sequence IDs 1302-1303 are disclosed in order of appearance. [Figure 2] A schematic diagram showing the structure and chemical modification pattern of another common GalNAc-conjugated PNPLA3 oligonucleotide is disclosed. [Modes for carrying out the invention]

[0050] overview ALD and NAFLD are serious public health burdens. ALD and NAFLD are chronic liver disorders that begin with the accumulation of triglycerides (TG) in the liver (steatosis) and progress to hepatitis and fibrosis, cirrhosis, and even liver cancer. PNPLA3 148M is a genetic factor that has been shown to be associated not only with ALD and NAFLD, but also with cirrhosis, HCC, and liver-related death.

[0051] RNA interference (RNAi) is a process that involves introducing exogenous RNA into a cell to induce specific degradation of mRNA encoding a target protein, thereby reducing the expression of the target gene.

[0052] In humans, PNPLA3 is 481 amino acids long and has a predicted molecular weight of 52.865 kD. Exemplary nucleic acid sequences of PNPLA3 can be found in NCBI reference sequence numbers NM_025225 (human), NM_054088 or XM_006520346 (mouse), NM_001282324 (rat), XM_015457081 (primate), XM_005567051 (primate), and XM_001109144 (primate). However, those skilled in the art will understand that further examples of PNPLA3 mRNA sequences are readily available using publicly available databases such as GenBank and UniProt.

[0053] definition As used herein, “approximately” means within a statistically meaningful range of a given value, such as concentration, length, molecular weight, pH, sequence similarity, time frame, temperature, or volume. Such a value or range may be within one order of magnitude of a given value or range, typically within 20%, more typically within 10%, and even more typically within 5%. The acceptable variation encompassed by “approximately” will vary depending on the system being tested and will be readily apparent to those skilled in the art.

[0054] As used herein, “administer,” “give administration,” and “give administration” refer to providing an individual with a substance (e.g., an oligonucleotide or composition herein) in a pharmacologically useful manner (e.g., to treat a disease, disorder, or condition of the individual).

[0055] As used herein, “antisense strand” means the oligonucleotide herein that is complementary to the region of the target sequence. Similarly, as used herein, “sense strand” means the oligonucleotide herein that is complementary to the region of the antisense strand.

[0056] As used herein, “asialoglycoprotein receptor” or “ASGPR” refers to a bipartite type C lectin formed by a larger 48 kDa subunit (ASGPR-1) and a smaller 40 kDa subunit (ASGPR-2). ASGPR is primarily expressed on the sinusoidal surface of hepatocytes and plays a major role in the binding, internalization, and subsequent efflux of circulating glycoproteins (asialoglycoproteins) containing terminal galactose or GalNAc residues.

[0057] As used herein, “weakening,” “to reduce,” and “weakening” refer to reducing or effectively stopping. In non-limiting examples, one or more of the treatments herein may reduce or effectively stop the onset or progression of AH, ALD, CCA, PSC, cirrhosis, HCC, NAFLD, and NASH, and related diseases, disorders, and conditions in an individual. This attenuation may be exemplified, for example, by a reduction in one or more aspects of AH, ALD, cirrhosis, HCC, CCA, PSC, NAFLD, and NASH, and related diseases, disorders, and conditions (e.g., symptoms, tissue characteristics, and cellular activity, inflammatory activity, or immune activity); the absence of detectable progression (worsening) of one or more aspects of AH, ALD, cirrhosis, HCC, CCA, PSC, NAFLD, and NASH, and related diseases, disorders, and conditions, or the absence of detectable aspects of AH, ALD, cirrhosis, HCC, NAFLD, and NASH, and related diseases, disorders, and conditions in an individual where such a pattern would otherwise be expected.

[0058] As used herein, “attenuate,” “to reduce,” and “attenuate” mean to reduce or effectively halt. In non-limiting examples, one or more of the treatments herein may reduce or effectively halt the onset or progression of dyslipidemia / hypertriglyceridemia / hyperlipidemia in a subject. This attenuation may be exemplified, for example, by a reduction in one or more aspects of AH, ALD, cirrhosis, HCC, CCA, PSC, NAFLD, and NASH, and related diseases, disorders, and conditions (e.g., symptoms, tissue characteristics, and cellular activity, inflammatory activity, or immune activity) that would otherwise be expected.

[0059] As used herein, “complementary” means a structural relationship between two nucleotides that enables them to form base pairs with one another (for example, on two opposing nucleic acids or on opposing regions of a single nucleic acid chain). For example, a purine nucleotide of one nucleic acid that is complementary to a pyrimidine nucleotide of an opposing nucleic acid may base pair with one another by forming a hydrogen bond with one another. Complementary nucleotides may base pair in the Watson-Crick configuration or in any other form that enables the formation of a stable double helix. Similarly, two nucleic acids may have regions of multiple nucleotides that form a complementary region by being complementary to one another, as described herein.

[0060] As used herein, “to come into contact with,” “to come into contact with,” etc., means introducing or delivering RNAi directly or indirectly to a cell, for example, by promoting or effectively facilitating its uptake or absorption into the cell.

[0061] As used herein, “deoxyribonucleotide” means a nucleotide that, compared to a ribonucleotide, has a hydrogen atom instead of a hydroxyl atom at the 2' position of its pentose sugar. A modified deoxyribonucleotide has one or more modification substitutions on atoms other than the 2' position, for example, a modification or substitution in the nucleic acid base, sugar, or phosphate group, or a modification or substitution of the nucleic acid base, sugar, or phosphate group.

[0062] As used herein, “double-stranded oligonucleotide” or “ds oligonucleotide” means an oligonucleotide that is substantially in a double-stranded form. Complementary base pairing of the double-stranded region(s) of a ds oligonucleotide may occur between antiparallel sequences of nucleotides from covalently separated nucleic acid strands. Similarly, complementary base pairing of the double-stranded region(s) of a ds oligonucleotide may occur between antiparallel sequences of nucleotides from covalently bonded nucleic acid strands. Furthermore, complementary base pairs of the double-stranded region(s) of a ds oligonucleotide may occur from a single nucleic acid strand that provides a complementary antiparallel sequence of nucleotides that fold together (e.g., via a hairpin) to form base pairs. A ds oligonucleotide may include two covalently separated nucleic acid strands that are fully double-stranded. However, a ds oligonucleotide may include two covalently separated nucleic acid strands that are partially double-stranded (e.g., having overhangs at one or both ends). ds oligonucleotides can contain antiparallel sequences of partially complementary nucleotides and therefore may have one or more mismatches, which may include internal or terminal mismatches.

[0063] As used herein, with respect to nucleic acids (e.g., oligonucleotides), “double helix” means a structure formed by complementary base pairing of two antiparallel sequences of nucleotides.

[0064] As used herein, “excipient” means a non-therapeutic agent that can be included in the compositions herein, for example, to give or contribute to a desired consistency or stabilizing effect.

[0065] As used herein, “hepatocytes” refers to the cells of the parenchymal tissue of the liver. These cells constitute approximately 70%–85% of the liver’s mass and produce serum albumin, fibronectin (FBN), and the prothrombin group of coagulation factors (excluding factors 3 and 4). Markers of hepatocyte-lineage cells include, but are not limited to, transthyretin (Ttr), glutamine synthetase (Glul), hepatocyte nuclear factor 1a (Hnf1a), and hepatocyte nuclear factor 4a (Hnf4a). Markers of mature hepatocytes may include, but are not limited to, cytochrome P450 (Cyp3a11), fumarylacetoacetate hydrolase (Fah), glucose 6-phosphate (G6p), albumin (Alb), and OC2-2F8. See, for example, Huch et al. (2013) Nature 494:247–50.

[0066] As used herein, “hepatotoxic agent” means a compound, virus, or other substance that is toxic to the liver in itself or can be processed to form metabolites that are toxic to the liver. Examples of hepatotoxic agents include, but are not limited to, carbon tetrachloride (CCl4), acetaminophen (paracetamol), vinyl chloride, arsenic, chloroform, and nonsteroidal anti-inflammatory drugs (such as aspirin and phenylbutazone).

[0067] As used herein, “unstable linker” means a linker that can be cleaved (for example, by an acidic pH). Similarly, “fairly stable linker” means a linker that cannot be cleaved.

[0068] As used herein, “inflammation of the liver” or “hepatitis” means a physical condition characterized by swelling, dysfunction, and / or pain in the liver, particularly as a result of injury or infection, which may be caused by exposure to hepatotoxic agents. Symptoms may include jaundice, fatigue, weakness, nausea, vomiting, loss of appetite, and weight loss. If left untreated, inflammation of the liver may progress to fibrosis, cirrhosis, liver failure, or liver cancer.

[0069] As used herein, “hepatic fibrosis,” “liver fibrosis,” or “fibrosis of the liver” refers to an excessive accumulation of extracellular matrix proteins in the liver, which may include collagen (I, III, and IV), FBN, undulin, elastin, laminin, hyaluronan, and proteoglycans, resulting from inflammation and hepatic cell death. If left untreated, hepatic fibrosis can progress to cirrhosis, liver failure, or liver cancer.

[0070] As used herein, “loop” means an unpaired region of a nucleic acid (e.g., oligonucleotide) adjacent to two sufficiently complementary antiparallel regions of the nucleic acid, where, under appropriate hybridization conditions (e.g., in a phosphate buffer, intracellularly), the two antiparallel regions adjacent to the unpaired region hybridize to form a double helix (called a “stem”).

[0071] As used herein, “modified nucleotide bond” means a nucleotide bond having one or more chemical modifications compared to a reference nucleotide bond having a phosphodiester bond. Modified nucleotides may be bonds that do not exist in nature. Typically, modified nucleotide bonds confer one or more desirable properties to the nucleic acid in which they are present. For example, modified nucleotides can improve thermal stability, resistance to degradation, nuclease resistance, solubility, bioavailability, biological activity, and immunogenicity.

[0072] As used herein, “modified nucleotide” refers to a nucleotide having one or more chemical modifications compared to a corresponding reference nucleotide selected from adenine ribonucleotide, guanine ribonucleotide, cytosine ribonucleotide, uracil ribonucleotide, adenine deoxyribonucleotide, guanine deoxyribonucleotide, cytosine deoxyribonucleotide, and thymidine deoxyribonucleotide. Modified nucleotides may be nucleotides that do not exist in nature. Modified nucleotides may have one or more chemical modifications to their sugar, nucleic acid base, and / or phosphate group. Additionally, or alternatively, modified nucleotides may have one or more chemical moieties conjugated to the corresponding reference nucleotide. Typically, modified nucleotides confer one or more desirable properties to the nucleic acid in which they are present. For example, modified nucleotides can improve thermal stability, resistance to degradation, nuclease resistance, solubility, bioavailability, biological activity, and reduced immunogenicity.

[0073] As used herein, “nicked tetraloop structure” means an RNAi oligonucleotide structure characterized by a separate sense (passenger) strand and an antisense (guide) strand, wherein the sense strand has a region complementary to the antisense strand, and at least one of the strands (generally the sense strand) has a tetraloop configured to stabilize an adjacent stem region formed within that at least one strand.

[0074] As used herein, “nucleotide” means a nucleoside (e.g., nucleic acid bases such as adenine, cytosine, guanine, thymine, or uracil) and a pentose (e.g., ribose or 2'-deoxyribose) and an organic molecule having a phosphate group, which can function as monomeric units of nucleic acid polymers such as deoxyribonucleic acid (DNA) and ribonucleic acid (RNA).

[0075] As used herein, “oligonucleotide” means a short nucleic acid molecule (e.g., less than about 100 nucleotides in length). Oligonucleotides may be single-stranded (ss) or ds. Oligonucleotides may or may not have a double-stranded region. A non-limiting set of examples of oligonucleotides may include, but are not limited to, small interfering RNA (siRNA), microRNA (miRNA), short hairpin RNA (shRNA), dicer substrate interfering RNA (dsiRNA), antisense oligonucleotide (ASO), short siRNA, or ss siRNA. Typically, ds oligonucleotides are RNAi oligonucleotides.

[0076] As used herein, “overhang” means a terminal unpaired nucleotide(s) resulting from one strand or region extending beyond the end of the complementary strand with which that strand or region forms a double helix. An overhang may include one or more unpaired nucleotides extending from the double-stranded region at the 5' or 3' end of a ds oligonucleotide. An overhang may be a 3' or 5' overhang on the antisense or sense strand of a ds oligonucleotide.

[0077] As used herein, “phosphate analog” means a chemical moiety that mimics the electrostatic and / or steric properties of a phosphate group. In some embodiments, the phosphate analog is located at the 5' terminal nucleotide of an oligonucleotide in place of a 5'-phosphate that is susceptible to enzymatic removal. The 5'-phosphate analog may include a phosphatase-resistant binding. Examples of phosphate analogs include, but are not limited to, 5'-phosphonates such as 5'-methylenephosphonate (5'-MP) and 5'-(E)-vinylphosphonate (5'-VP). Oligonucleotides may have a phosphate analog at the 4' position of the sugar carbon in the 5' terminal nucleotide (referred to as a 4'-phosphate analog). Examples of 4'-phosphate analogs include oxymethylphosphonates or analogs thereof, in which the oxygen atom of the oxymethyl group is bonded to the sugar moiety (e.g., its 4'-carbon). See, for example, International Patent Application Publication WO2018 / 045317. Other modifications to the 5' end of oligonucleotides have been developed (see, for example, International Patent Application Publication WO2011 / 133871; U.S. Patent No. 8,927,513; and Prakash et al. (2015) Nucleic Acids Res. 43:2993-3011).

[0078] As used herein, “PNPLA3-related disease,” “PNPLA3-related disorder,” or “PNPLA3-related condition” means a condition having increased PNPLA3 expression and / or the presence of, for example, the PNPLA3 I148M variant. Exemplary PNPLA3-related conditions, diseases, or disorders include, but are not limited to, fat accumulation in the liver, cirrhosis, fatty liver (steatosis), hepatocyte necrosis, HCC, hepatic fibrosis, hepatic inflammation, NASH, NAFLD, or obesity.

[0079] As used herein, “decreased expression” with respect to a gene (e.g., PNPLA3) means a decrease in the amount or level of the RNA transcript (e.g., PNPLA3 mRNA) or protein encoded by the gene, and / or a decrease in the amount or level of the gene’s activity in the cell, cell population, sample, or subject, compared to a suitable reference (e.g., a reference cell, cell population, sample, or subject). For example, contacting cells with the oligonucleotides specified herein (e.g., oligonucleotides having an antisense strand with a nucleotide sequence complementary to the nucleotide sequence containing PNPLA3 mRNA) may result in a decrease in the amount or level of mRNA, protein, and / or activity (e.g., due to degradation of PNPLA3 mRNA via the RNAi pathway) compared to cells not treated with the ds oligonucleotide. Similarly, as used herein, “decreased expression” means an action that results in decreased expression of a gene (e.g., PNPLA3). Specifically, as used herein, “reduced PNPLA3 expression” means a decrease in the amount or level of PNPLA3 mRNA, PNPLA3 protein, and / or PNPLA3 activity in cells, cell populations, samples, or subjects compared to a suitable reference (e.g., reference cells, cell populations, samples, or subjects).

[0080] As used herein, “complementary region” means a sequence of nucleotides in a nucleic acid (e.g., ds-oligonucleotide) that is sufficiently complementary to the antiparallel sequence of nucleotides, thereby enabling hybridization between two nucleotide sequences under appropriate hybridization conditions (e.g., in a phosphate buffer, intracellularly, etc.). Oligonucleotides as used herein include target sequences having a region complementary to the mRNA target sequence.

[0081] As used herein, "ribonucleotide" means a nucleotide having ribose as its pentose sugar and a hydroxyl group at its 2' position. A modified ribonucleotide is a ribonucleotide having one or more modifications or substitutions on atoms other than the 2' position, for example, having modifications or substitutions in the nucleic acid base, sugar, or phosphate group, or having modifications or substitutions of the nucleic acid base, sugar, or phosphate group.

[0082] As used herein, “RNAi oligonucleotide” means either (a) a ds oligonucleotide having a sense strand (passenger) and an antisense strand (guide), wherein the antisense strand or a portion of the antisense strand is used to cleave target mRNA by Argonaut 2 (Ago2) endonuclease, or (b) an ss oligonucleotide having a single-stranded antisense strand, wherein the antisense strand (or a portion of the antisense strand) is used to cleave target mRNA by Ago2 endonuclease.

[0083] As used herein, “chain” refers to a single, continuous sequence of nucleotides linked together via internucleotide bonds (e.g., phosphodiester bonds or phosphorothioate bonds). A chain may have two free ends (e.g., a 5' end and a 3' end).

[0084] As used herein, “subject” means any mammal, including cats, dogs, mice, rats, and primates, in particular humans. Furthermore, “individual” or “patient” may be used interchangeably with “subject.”

[0085] As used herein, “synthesized” means nucleic acids or other molecules that are artificially synthesized (e.g., using machinery (e.g., a solid-phase nucleic acid synthesizer)) or that do not originate from natural sources (e.g., cells or organisms) that otherwise normally produce nucleic acids or molecules thereof.

[0086] As used herein, “targeting ligand” means a molecule (e.g., amino sugars, carbohydrates, cholesterol, lipids, or polypeptides) that selectively binds to a homologous molecule (e.g., a receptor) of a tissue or cell of interest and is conjugable to another substance for targeting that substance to the tissue or cell of interest. For example, a targeting ligand can be conjugated to an oligonucleotide as herein for the purpose of targeting the oligonucleotide to a specific tissue or cell of interest. A targeting ligand can selectively bind to a cell surface receptor. Thus, when conjugated to an oligonucleotide, a targeting ligand facilitates the delivery of the oligonucleotide to a specific cell via selective binding to a receptor expressed on the cell surface, as well as endosomal internalization by the cell of a complex containing the oligonucleotide, the targeting ligand, and the receptor. Furthermore, a targeting ligand can be conjugated to an oligonucleotide via a linker that is cleaved after or during internalization so that the oligonucleotide is released from the targeting ligand within the cell.

[0087] As used herein, “tetraloop” refers to a loop that increases the stability of adjacent double helices formed by the hybridization of adjacent nucleotide sequences. This increased stability reduces the melting temperature (T) of the adjacent stem double helices. m This can be detected as an increase in the T of adjacent stem double helix, which is expected to be the average of a set of loops of equivalent length consisting of randomly selected nucleotide sequences. m Higher than. For example, a tetraloop is a hairpin containing a double helix of at least two base pairs (bp) in length, heated in 10 mM NaHPO4 at a temperature of at least about 50°C, at least about 55°C, at least about 56°C, at least about 58°C, at least about 60°C, at least about 65°C, or at least about 75°C. mThis can result in the tetraloop stabilizing the bp of adjacent stem double helixes through stacking interactions. Furthermore, interactions between nucleotides within a tetraloop include, but are not limited to, non-Watson-Crick base pairings, stacking interactions, hydrogen bonding, and contact interactions (Cheong et al. (1990) Nature 346:680-682; Heus & Pardi (1991) Science 253:191-194). In this specification, a tetraloop may or may contain about 3 to 6 nucleotides, typically about 4 to 5 nucleotides. Thus, a tetraloop can have 3, 4, 5, or 6 nucleotides, typically 4 nucleotides, which may or may not be modified (e.g., may or may not be conjugated to the targeting moiety). In tetraloops, any nucleotide can be used, and such nucleotides can be given standard IUPAC-IUB notation as described in Cornish-Bowden (1985) Nucleic Acids Res. 13:3021-30. For example, the letter "N" can be used to mean that any base can be in that position, the letter "R" can be used to indicate that A (adenine) or G (guanine) can be in that position, and "B" can be used to indicate that C (cytosine), G (guanine), or T (thymine) can be in that position. Examples of tetraloops include the tetraloop of the UNCG family (e.g., UUCG), the tetraloop of the GNRA family (e.g., GAAA), and the CUUG tetraloop (Woese et al. (1990) Proc. Natl. Acad. Sci. USA 87:8467-71; Antao et al. (1991) Nucleic Acids Res. 19:5901-05).Examples of DNA tetraloops include the d(GNNA) family of tetraloops (e.g., d(GTTA)), the d(GNRA) family of tetraloops, the d(GNAB) family of tetraloops, the d(CNNG) family of tetraloops, and the d(TNCG) family of tetraloops (e.g., d(TTCG)). See, for example, Nakano et al. (2002) Biochem. 41:4281-92; and Shinji et al. (2000) Nippon Kagakkai Koen Yokoshu 78:731. In this specification, tetraloops can be found within nicked tetraloop structures.

[0088] As used herein, “to treat” or “to treat” means the act of providing care to an individual in need of care by administering a therapeutic agent (e.g., oligonucleotides as herein) to the individual for the purpose of improving the individual’s health and / or living condition with respect to an existing condition (e.g., disease, disability) or to prevent or reduce the likelihood of the condition developing. Treatment may include reducing the frequency or severity of at least one sign, symptom, or contributing factor of a condition (e.g., disease, disability) experienced by the individual.

[0089] As used herein, "iRNA," "iRNA agent," "RNAi," "RNAi agent," and "RNA interferant" refer to substances containing RNA that mediate targeted cleavage of RNA transcripts via the RNA-induced silencing complex (RISC) pathway. iRNA directs sequence-specific degradation of mRNA through RNA interference. iRNA modulates, inhibits, or reduces intracellular PNPLA3 expression.

[0090] composition In some embodiments, the disclosure provides oligonucleotides (e.g., double-stranded RNAi oligonucleotides) that reduce, regulate, or inhibit PNPLA3 expression in the liver. In some embodiments, the oligonucleotides provided herein are used to treat diseases associated with PNPLA3 expression. In some embodiments, the disclosure provides a method for treating diseases associated with PNPLA3 expression by reducing, regulating, or inhibiting PNPLA3 expression in the liver (e.g., cells constituting the liver).

[0091] Oligonucleotide inhibitors of PNPLA3 expression I. PNPLA3 Target Sequence: The oligonucleotides described herein (e.g., RNAi oligonucleotides) target a target sequence containing PNPLA3 mRNA (i.e., a PNPLA3 target sequence). In some embodiments, the oligonucleotide or a portion, fragment, or chain thereof (e.g., the antisense or guide strand of a dsRNAi oligonucleotide) binds to or anneals to the PNPLA3 target sequence, thereby inhibiting PNPLA3 expression. In some embodiments, the oligonucleotide targets the PNPLA3 target sequence and inhibits PNPLA3 expression in vivo. In some embodiments, the amount or degree of PNPLA3 expression inhibition by the oligonucleotide targeting the PNPLA3 target sequence correlates with the potency of the oligonucleotide. In some embodiments, the amount or degree of PNPLA3 expression inhibition by the oligonucleotide targeting the PNPLA3 target sequence correlates with the amount or degree of therapeutic effect in an individual having or suspected of having a disease, disorder, or condition related to PNPLA3 expression treated with the oligonucleotide.

[0092] Through the examination and analysis of nucleotide sequences of PNPLA3 mRNA, including mRNA from multiple different species (e.g., human, cynomolgus monkey, and rhesus monkey; see, e.g., Example 1), and as a result of in vitro and in vivo studies (see, e.g., Examples 2-3), it is shown herein that certain nucleotide sequences of PNPLA3 mRNA are more susceptible to oligonucleotide-based PNPLA3 expression inhibition than other nucleotide sequences and are therefore useful as target sequences for the oligonucleotides herein. In some embodiments, the sense strand of an oligonucleotide (e.g., dsRNAi oligonucleotide) described herein (e.g., Table 1 or 3, or Tables A, B, C, or D) contains a PNPLA3 target sequence. In some embodiments, part or region of the sense strand of an oligonucleotide described herein (e.g., Table 1 or 3, or Tables A, B, C, or D) contains a PNPLA3 target sequence. In some embodiments, the PNPLA3 target sequence contains or consists of one of sequence numbers 1167-1176.

[0093] II. PNPLA3 mRNA Target Sequence: In some embodiments, the oligonucleotides herein have a complementary region to PNPLA3 mRNA (e.g., within the target sequence of PNPLA3 mRNA) for inhibiting PNPLA3 expression by targeting PNPLA3 mRNA in cells. In some embodiments, the oligonucleotides herein include a PNPLA3 target sequence (e.g., the antisense or guide strand of a ds oligonucleotide) having a complementary region that binds to or anneals to the PNPLA3 mRNA target sequence by complementary (Watson-Crick) base pairing. The target sequence or complementary region has a length and base content suitable for enabling the binding or annealing of the oligonucleotide (or its chain) to PNPLA3 mRNA and inhibiting its expression. In some embodiments, the target sequence or complementary region is at least about 12, at least about 13, at least about 14, at least about 15, at least about 16, at least about 17, at least about 18, at least about 19, at least about 20, at least about 21, at least about 22, at least about 23, at least about 24, at least about 25, at least about 26, at least about 27, at least about 28, at least about 29, or at least about 30 nucleotides long. Alternatively, the target sequence or complementary region is about 12 to about 30 nucleotides long (e.g., 12 to 30, 12 to 22, 15 to 25, 17 to 21, 18 to 27, 19 to 27, or 15 to 30). Alternatively, the target sequence or complementary region is approximately 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 nucleotides long. In certain embodiments, the target sequence or complementary region is 18 nucleotides long. In certain embodiments, the target sequence or complementary region is 19 nucleotides long. In certain embodiments, the target sequence or complementary region is 20 nucleotides long. In certain embodiments, the target sequence or complementary region is 21 nucleotides long. In certain embodiments, the target sequence or complementary region is 22 nucleotides long. In certain embodiments, the target sequence or complementary region is 23 nucleotides long. In certain embodiments, the target sequence or complementary region is 24 nucleotides long.

[0094] In some embodiments, the oligonucleotides herein include a target sequence or complementary region (e.g., an antisense strand or guide strand of a double-stranded oligonucleotide) that is fully complementary to the PNPLA3 target sequence. In some embodiments, the target sequence or complementary region is partially complementary to the PNPLA3 target sequence. In some embodiments, the oligonucleotide includes a target sequence or complementary region that is fully complementary to any one of the sequences of SEQ ID NOs. 1167-1176. In some embodiments, the oligonucleotide includes a target sequence or complementary region that is partially complementary to any one of the sequences of SEQ ID NOs. 1167-1176.

[0095] Alternatively, in some embodiments, the oligonucleotides herein include a target sequence or complementary region complementary to a sequence of nucleotides constituting PNPLA3 mRNA, the sequence of nucleotides being about 12 to about 30 nucleotides long (e.g., 12 to 30, 12 to 28, 12 to 26, 12 to 24, 12 to 20, 12 to 18, 12 to 16, 14 to 22, 16 to 20, 18 to 20, or 18 to 19 nucleotides long). In some embodiments, the oligonucleotides include a target sequence or complementary region complementary to a sequence of nucleotides constituting PNPLA3 mRNA, the sequence of nucleotides being 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 nucleotides long. In some embodiments, the oligonucleotides include a target sequence or complementary region complementary to a sequence of nucleotides constituting PNPLA3 mRNA, the sequence of nucleotides being 19 nucleotides long. In some embodiments, the oligonucleotide comprises a target sequence or complementary region complementary to a sequence of nucleotides constituting the PNPLA3 mRNA, wherein the sequence of nucleotides is 20 nucleotides long. In other embodiments, the oligonucleotide comprises a target sequence or complementary region complementary to a sequence of nucleotides from any one of sequence numbers 1167-1176, wherein the sequence of nucleotides is optionally 19 nucleotides long.

[0096] With respect to the target sequence or complementary region of the oligonucleotides herein, it is complementary to the sequence of nucleotides of the sequence described in any one of SEQ ID NOs. 1167-1176 and extends over the entire length of the antisense strand. In some embodiments, the complementary region of the oligonucleotide is complementary to the sequence of nucleotides of the sequence described in any one of SEQ ID NOs. 1167-1176 and extends over a portion of the entire length of the antisense strand. In some additional embodiments, the oligonucleotide includes a complementary region (e.g., on the antisense strand of the ds oligonucleotide) that is at least partially (e.g., fully) complementary to a sequence of nucleotides over nucleotides 1-20 of the sequence described in any one of SEQ ID NOs. 1167-1176.

[0097] Alternatively, the oligonucleotides herein include a target sequence or complementary region having one or more base pair (bp) mismatches with the corresponding PNPLA3 target sequence. In some embodiments, the target sequence or complementary region has up to approximately one, two, three, four, or five mismatches with the corresponding PNPLA3 target sequence, as long as the ability of the target sequence or complementary region to bind to or anneal to PNPLA3 mRNA and / or the ability of the oligonucleotide to reduce or inhibit PNPLA3 expression is maintained under appropriate hybridization conditions. In other words, the target sequence or complementary region has one or fewer, two or fewer, three or fewer, four or fewer, or five or fewer mismatches with the corresponding PNPLA3 target sequence, as long as the ability of the target sequence or complementary region to bind to or anneal to PNPLA3 mRNA and / or the ability of the oligonucleotide to reduce or inhibit PNPLA3 expression is maintained under appropriate hybridization conditions. In some embodiments, the oligonucleotide includes a target sequence or complementary region having one mismatch with the corresponding target sequence. In some embodiments, the oligonucleotide includes a target sequence or complementary region having two mismatches with the corresponding target sequence. In some embodiments, the oligonucleotide includes a target sequence or complementary region having three mismatches with the corresponding target sequence. In some embodiments, the oligonucleotide includes a target sequence or complementary region having four mismatches with the corresponding target sequence. In some embodiments, the oligonucleotide includes a target sequence or complementary region having five mismatches with the corresponding target sequence. In other embodiments, the oligonucleotide includes a target sequence or complementary region with two or more mismatches (e.g., 2, 3, 4, 5 or more mismatches) with the corresponding target sequence, where at least two (e.g., all) of the mismatches are contiguous (e.g., 2, 3, 4, 5 or more mismatches are contiguous), or the mismatches are scattered at any position throughout the target sequence or complementary region.In other embodiments, the oligonucleotide comprises a target sequence or complementary region with two or more mismatches (e.g., 2, 3, 4, 5 or more mismatches) with the corresponding target sequence, where at least two (e.g., all) of the mismatches are contiguous (e.g., 2, 3, 4, 5 or more mismatches are contiguous), or at least one non-mismatched base pair is located between the mismatches, or a combination thereof.

[0098] III. Types of Oligonucleotides: Various oligonucleotide types and / or structures, including but not limited to RNAi oligonucleotides, antisense oligonucleotides, and miRNAs, are useful for targeting PNPLA3 mRNA. Any oligonucleotide type described herein or elsewhere is intended to be used as a framework for incorporating the target sequences herein for the purpose of inhibiting PNPLA3 expression. In some embodiments, the oligonucleotides herein inhibit PNPLA3 expression by being incorporated into an RNAi pathway upstream or downstream of Dicer involvement. For example, RNAi oligonucleotides have been developed with each strand having a size of approximately 19–25 nucleotides and at least one 3' end overhang of 1–5 nucleotides (see, e.g., U.S. Patent No. 8,372,968). Longer oligonucleotides have also been developed that are processed by Dicer to produce active RNAi products (see, e.g., U.S. Patent No. 8,883,996). Further work has resulted in the creation of extended ds oligonucleotides in which at least one end of at least one chain extends beyond the target region of the double helix, such as structures in which one chain contains a thermodynamically stable tetraloop structure (see, for example, U.S. Patents 8,513,207 and 8,927,705, and International Patent Application Publication WO2010 / 033225). Such structures include ss extensions (on one or both sides of the molecule) and ds extensions.

[0099] The oligonucleotides described herein are incorporated into RNAi pathways downstream of Dicer involvement (e.g., Dicer cleavage). In some embodiments, the oligonucleotide has an overhang (e.g., 1, 2, or 3 nucleotides long) at the 3' end of the sense strand. In some embodiments, the oligonucleotide (e.g., siRNA) comprises a 21-nucleotide guide strand that is antisense to the target RNA (e.g., PNPLA3 mRNA) and a complementary passenger strand, both strands annealing to form a 19 bp double helix with a 2-nucleotide overhang at either or both 3' ends. Longer oligonucleotide designs have also been considered, including oligonucleotides having a 23-nucleotide guide strand and a 21-nucleotide passenger strand, in which case there is a blunt end on the right side of the molecule (3' end of passenger strand / 5' end of guide strand) and a 2-nucleotide 3' guide strand overhang on the left side of the molecule (5' end of passenger strand / 3' end of guide strand). In such molecules, a 21 bp double-stranded region is present. See, for example, U.S. Patent No. 9,012,138; No. 9,012,621; and No. 9,193,753.

[0100] The oligonucleotides herein comprise a sense strand and an antisense strand, both in the range of approximately 17 to approximately 26 nucleotide lengths (e.g., 17 to 26, 20 to 25, or 21 to 23). In some embodiments, the oligonucleotide comprises a sense strand and an antisense strand, both in the range of approximately 19 to approximately 22 nucleotide lengths. In some embodiments, the sense strand and the antisense strand are of equal length. In some embodiments, the oligonucleotide comprises a sense strand and an antisense strand such that a 3' overhang exists on either the sense strand or the antisense strand, or on both the sense strand and the antisense strand. In some examples, for oligonucleotides having a sense strand and an antisense strand both in the range of approximately 21 to approximately 23 nucleotide lengths, the 3' overhang on the sense strand, the antisense strand, or on both the sense strand and the antisense strand is 1 or 2 nucleotides long. In some embodiments, the oligonucleotide comprises a 22-nucleotide guide strand and a 20-nucleotide passenger strand, in which case there is a blunt end on the right side of the molecule (3' end of the passenger strand / 5' end of the guide strand) and a 2-nucleotide 3' guide strand overhang on the left side of the molecule (5' end of the passenger strand / 3' end of the guide strand). In such molecules, there is a 20 bp double-stranded region.

[0101] Other oligonucleotide designs for use herein include: 16-mer siRNA (see, e.g., "Nucleic Acids in Chemistry & Biology", Blackburn (ed.), Royal Society of Chemistry, 2006), shRNA (e.g., with a stem of 19 bp or shorter; see, e.g., Moore et al. (2010) Methods Mol. Biol. 629:141-58), blunt siRNA (e.g., 19 bp long; see, e.g., Kraynack & Baker (2006) RNA 12:163-76), asymmetric siRNA (aiRNA; see, e.g., Sun et al. (2008) Nat. Biotechnol. 26:1379-82), asymmetric short double-stranded siRNA (see, e.g., Chang et al. (2009) Mol. Ther. 17:725-32), and fork siRNA (e.g., Hohjoh (2004) FEBS). Examples of oligonucleotide structures that may be used herein to reduce or inhibit PNPLA3 expression include ss siRNA (see, e.g., Elsner (2012) Nat. Biotechnol. 30:1063), dumbbell-shaped circular siRNA (see, e.g., Abe et al. (2007) J.Am. Chem. Soc. 129:15108-09), and small internal segmented interfering RNA (sisiRNA; see, e.g., Bramsen et al. (2007) Nucleic Acids Res. 35:5886-97). Further non-limiting examples of oligonucleotide structures that may be used herein to reduce or inhibit PNPLA3 expression include miRNA, shRNA, and short-chain siRNA (see, e.g., Hamilton et al. (2002) EMBO J. 21:4671-79; also see U.S. Patent Application Publication No. 2009 / 0099115).

[0102] Alternatively, the oligonucleotides herein are single-stranded (ss). Examples of such structures include, but are not limited to, ssRNAi molecules. Recent efforts have demonstrated the activity of ssRNAi molecules (see, e.g., Matsui et al. (2016) Mol.Ther. 24:946-55). In some embodiments, the oligonucleotide is an ASO. An ASO is an ss oligonucleotide that has been appropriately modified to have a nucleic acid base sequence containing the reverse complement of a target segment of a particular nucleic acid when written or drawn in the 5' to 3' direction, and to induce RNaseH-mediated cleavage of its target RNA in a cell (e.g., as a gapmer) or to inhibit translation of the target mRNA in a cell (e.g., as a mixmer). ASOs for use in this disclosure are modified in any suitable manner known in the art, including, for example, those shown in U.S. Patent No. 9,567,587 (e.g., modifications of length, sugar moieties of nucleic acid bases (pyrimidines, purines), and heterocyclic moieties of nucleic acid bases). Furthermore, ASOs have been used for decades to reduce the expression of specific target genes (see, for example, Bennett et al. (2017) Annu. Rev. Pharmacol. 57:81-105).

[0103] IV. Double-stranded RNAi oligonucleotides: ds oligonucleotides comprising a sense strand (i.e., passenger strand) and an antisense strand (i.e., guide strand) for targeting PNPLA3 mRNA and inhibiting PNPLA3 expression (e.g., via the RNAi pathway). In some embodiments, the sense strand and antisense strand are separate strands and not covalently bonded. In some embodiments, the sense strand and antisense strand are covalently bonded.

[0104] In some embodiments, the sense strand comprises a first region (R1) and a second region (R2), where R2 comprises a first sub-region (S1), a tetraloop (L) or triloop (triL), and a second sub-region (S2), where L or triL is located between S1 and S2, and S1 and S2 form a second double strand (D2). D2 has varying lengths. In some embodiments, D2 is about 1 to about 6 bp long. In some embodiments, D2 is 2 to 6, 3 to 6, 4 to 6, 5 to 6, 1 to 5, 2 to 5, 3 to 5, or 4 to 5 bp long. In other embodiments, D2 is 1, 2, 3, 4, 5, or 6 bp long. In certain embodiments, D2 is 6 bp long.

[0105] In some embodiments, the sense strand R1 and the antisense strand form a first double helix (D1). In some embodiments, D1 has a nucleotide length of at least about 15 (e.g., at least 15, at least 16, at least 17, at least 18, at least 19, at least 20, or at least 21). In some embodiments, D1 is about 12 to about 30 nucleotides long (e.g., 12 to 30, 12 to 27, 15 to 22, 18 to 22, 18 to 25, 18 to 27, 18 to 30, or 21 to 30 nucleotides). In other embodiments, D1 is at least 12 nucleotides long (e.g., at least 12, at least 15, at least 20, at least 25, or at least 30 nucleotides). In other embodiments, D1 is 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 nucleotides long. In certain embodiments, D1 is 20 nucleotides long. In some examples, D1 does not extend the entire length of the sense strand and / or antisense strand. In some examples, D1 extends the entire length of either the sense strand or the antisense strand, or both. In certain examples, D1 extends the entire length of both the sense strand and the antisense strand.

[0106] In certain examples, the ds oligonucleotides of this specification are as shown in Table 1: SEQ ID NOs: 9, 11, 13, 15, 17, 19, 21, 23, 25, 27, 29, 31, 33, 35, 37, 39, 41, 43, 45, 47, 49, 51, 53, 55, 57, 59, 61, 63, 65, 67, 69, 71, 73, 75, 77, 79, 81, 83, 85, 87, 89, 91, 93, 95, 97, 99, 101, 103, 105, 107, 109, 111, 113, 115, 117, 119, 121, 123, 125, 127, 129, 131, 133, 13 5, 137, 139, 141, 143, 145, 147, 149, 151, 153, 155, 157, 159, 161, 163, 165, 167, 169, 171, 173, 175, 177, 179, 181, 183, 185, 187, 189, 191, 193, 195, 197 ,199,201,203,205,207,209,211,213,215,217,219,221,223,225,227,229,231,233,235,237,239,241,243,245,247,249,251,253,255,257,259, 261, 263, 265, 267, 269, 271, 273, 275, 277, 279, 281, 283, 285, 287, 289, 291, 293, 295, 297, 299, 301, 303, 305, 307, 309, 311, 313, 315, 317, 319, 321, 3 23, 325, 327, 329, 331, 333, 335, 337, 339, 341, 343, 345, 347, 349, 351, 353, 355, 357, 359, 361, 363, 365, 367, 369, 371, 373, 375, 377, 379, 381, 383, 38 5, 387, 389, 391, 393, 395, 397, 399, 401, 403, 405, 407, 409, 411, 413, 415, 417, 419, 421, 423, 425, 427, 429, 431, 433, 435, 437, 439, 441, 443, 445, 447 ,449,451,453,455,457,459,461,463,465,467,469,471,473,475,477,479,481,483,485,487,489,491,493,495,497,499,501,503,505,507,509,511, 513, 515, 517, 519, 521, 523, 525, 527, 529, 531, 533, 535, 537, 539, 541, 543, 545, 547, 549, 551, 553, 555, 557, 559, 561, 563, 565, 567, 569, 571, 5 73, 575, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, 629, 631, 633, 63 5, 637, 639, 641, 643, 645, 647, 649, 651, 653, 655, 657, 659, 661, 663, 665, 667, 669, 671, 673, 675, 677, 679, 681, 683, 685, 687, 689, 691, 693, 695, 697 ,699,701,703,705,707,709,711,713,715,717,719,721,723,725,727,729,731,733,735,737,739,741,743,745,747,749,751,753,755,757,759, A sense strand having one of the sequences 761, 763, 765, 767, 769, 771, 773, and 775, and sequence numbers 10, 12, 14, 16, 18, 20, 22, 24, 26, 28, 30, 32, 34, 36, 38, 40, 42, 44, 46, 48, 50, 52, 54, 56, 58, 60, 62, 64, 66, 68, 70, 72, 74, 76, 78, 80, 82, 84, 86, 88, 90, 92, 94, 96, 98, 100, 102, 104, 106, 108, 110, 112, 114, 116, 118, 120, 122, 124, 126, 12 8, 130, 132, 134, 136, 138, 140, 142, 144, 146, 148, 150, 152, 154, 156, 158, 160, 162, 164, 166, 168, 170, 172, 174, 176, 178, 180, 182, 184, 186, 188, 190 ,192,194,196,198,200,202,204,206,208,210,212,214,216,218,220,222,224,226,228,230,232,234,236,238,240,242,244,246,248,250,252,254、256、258、260、262、264、266、268、270、272、274、276、278、280、282、284、286、288、290、292、294、296、298、300、302、304、306、308、310、312、314、316、318、320、322、324、326、328、330、332、334、336、338、340、342、344、346、348、350、352、354、356、358、360、362、364、366、368、370、372、374、376、378、380、382、384、386、388、390、392、394、396、398、400、402、404、406、408、410、412、414、416、418、420、422、424、426、428、430、432、434、436、438、440、442、444、446、448、450、452、454、456、458、460、462、464、466、468、470、472、474、476、478、480、482、484、486、488、490、492、494、496、498、500、502、504、506、508、510、512、514、516、518、520、522、524、526、528、530、532、534、536、538、540、542、544、546、548、550、552、554、556、558、560、562、564、566、568、570、572、574、576、578、580、582、584、586、588、590、592、594、596、598、600、602、604、606、608、610、612、614、616、618、620、622、624、626、628、630、632、634、636、638、640、642、644、646、648、650、652、654、656、658、660、662、664、666、668、670、672、674、676、678、680、682、684、686、688、690、692、694、696、698、700、702、704、706、708、710、712、714、716、718、720、722、724、726、728、730、732、734、736、738、740、742、744、746、748、750、752、It includes an antisense chain having a complementary sequence selected from 754, 756, 758, 760, 762, 764, 766, 768, 770, 772, 774, and 776. Alternatively, the ds oligonucleotides of this specification are as listed in Table 3: SEQ ID NOs: 777, 779, 781, 783, 785, 787, 789, 791, 793, 795, 797, 799, 801, 803, 805, 807, 809, 811, 813, 815, 817, 819, 821, 823, 825, 827, 829, 831, 833, 835, 837, 839, 841, 843, 845, 847, 849, 851, 853, 855, 857, 859, 861, 863, 865, 867, 869, 8 71, 873, 875, 877, 879, 881, 883, 885, 887, 889, 891, 893, 895, 897, 899, 901, 903, 905, 907, 909, 911, 913, 915, 917, 919, 921, 923, 925, 927, 929, 931, 933, 935, 937, 939, 941, 943, 945, 947, 949, 951, 953, 955, 957, 959, 961, 963, 965, 967, 969, 971, 973, 975, 977, 979, 981, 983, 985 ,987,989,991,993,995,997,999,1001,1003,1005,1007,1009,1011,1013,1015,1017,1019,1021,1023,1025,1027,1029,1031,1033,1035,1037,1039,1041,1043,1045,1047,1049,1051,1053,1055,1057,1059,1061,1063,1065,1067,1069,1071,1073,1075,1077,1079,1 A sense strand having one of the sequences 081, 1083, 1085, 1087, 1089, 1091, 1093, 1095, 1097, 1099, 1101, 1103, 1105, 1107, 1109, 1111, 1113, 1115, 1117, 1119, 1121, 1123, 1125, 1127, 1129, 1131, 1133, 1135, 1137, 1139, 1141, 1143, 1145, 1147, 1149, 1151, 1153, 1155, 1157, 1159, 1161, and 1163,Sequence numbers 778, 780, 782, 784, 786, 788, 790, 792, 794, 796, 798, 800, 802, 804, 806, 808, 810, 812, 814, 816, 818, 820, 822, 824, 826, 828, 830, 832, 834, 836, 838, 840, 842, 844, 846, 848, 850, 852, 854, 856, 858, 860, 862, 864, 866, 868, 870, 872, 874, 876, 878, 880, 882, 884, 886 ,888,890,892,894,896,898,900,902,904,906,908,910,912,914,916,918,920,922,924,926,928,930,932,934,936,938,940,942,944,946,948,950,952,954,956,958,960,962,964,966,968,970,972,974,976,978,980,982,984,986,988,990,992,994,996,99 8, 1000, 1002, 1004, 1006, 1008, 1010, 1012, 1014, 1016, 1018, 1020, 1022, 1024, 1026, 1028, 1030, 1032, 1034, 1036, 1038, 1040, 1042, 1044, 1046, 1048, 1050, 1052, 1054, 1056, 1058, 1060, 1062, 1064, 1066, 1068, 1070, 1072, 1074, 1076, 1078, 1080, 1082, 1084, 1086, 1 It includes an antisense strand having a complementary sequence selected from 088, 1090, 1092, 1094, 1096, 1098, 1100, 1102, 1104, 1106, 1108, 1110, 1112, 1114, 1116, 1118, 1120, 1122, 1124, 1126, 1128, 1130, 1132, 1134, 1136, 1138, 1140, 1142, 1144, 1146, 1148, 1150, 1152, 1154, 1156, 1158, 1160, 1162, and 1164. In a specific example, the sense strand is one of sequence numbers 787, 843, 867, 871, 937, 1003, 1007, 1017, 1161, or 1163, and the antisense strand is sequence numbers 788, 844, 868, 872, 938,It is one of the following: 1004, 1008, 1018, 1162, or 1164.

[0107] In certain embodiments, this disclosure relates to sequences listed in Table A, B, C, or D (e.g., sequence numbers 1178, 1180, 1182, 1184, 1186, 1188, 1190, 1192, 1194, 1196, 1198, 1200, 1202, 1204, 1206, 1208, 1210, 1212, 1214, 1216, 1218, 1220, 1222, 1224, 1226, 1228, 1230, 1232, 1234, 1236, 1238, 1240, 1242 RNAi oligonucleotides for reducing or inhibiting PNPLA3 expression, which include a sense strand having (1244, 1246, 1248, 1250, 1252, 1254, 1256, 1258, 1260, 1262, 1264, 1266, 1268, 1270, 1272, 1274, 1276, 1278, 1280, 1282, 1284, 1286, 1288, 1290, 1292, 1294, 1296, 1298, or 1300), are described.

[0108] In certain embodiments, this disclosure relates to the sequences listed in Table A, B, C, or D (for example, sequence numbers 1179, 1181, 1183, 1185, 1187, 1189, 1191, 1193, 1195, 1197, 1199, 1201, 1203, 1205, 1207, 1209, 1211, 1213, 1215, 1217, 1219, 1221, 1223, 1225, 1227, 1229, 1231, 1233, 1235, 1237, 1239, 1241, 1243, RNAi oligonucleotides for reducing or inhibiting PNPLA3 expression, which include antisense strands having 1245, 1247, 1249, 1251, 1253, 1255, 1257, 1259, 1261, 1263, 1265, 1267, 1269, 1271, 1273, 1275, 1277, 1279, 1281, 1283, 1285, 1287, 1289, 1291, 1293, 1295, 1297, 1299, or 1301), are described. In certain embodiments, this disclosure relates to sequences listed in Table A, B, C, or D (e.g., sequence numbers 1178, 1180, 1182, 1184, 1186, 1188, 1190, 1192, 1194, 1196, 1198, 1200, 1202, 1204, 1206, 1208, 1210, 1212, 1214, 1216, 1218, 1220, 1222, 1224, 1226, 1228, 1230, 1232, 1234, 1236, 1238, 1240, 1242 RNAi oligonucleotides for reducing or inhibiting PNPLA3 expression, which include a sense strand having (1244, 1246, 1248, 1250, 1252, 1254, 1256, 1258, 1260, 1262, 1264, 1266, 1268, 1270, 1272, 1274, 1276, 1278, 1280, 1282, 1284, 1286, 1288, 1290, 1292, 1294, 1296, 1298, or 1300), are described.

[0109] In certain embodiments, this disclosure relates to the sequences listed in Table A, B, C, or D (for example, sequence numbers 1179, 1181, 1183, 1185, 1187, 1189, 1191, 1193, 1195, 1197, 1199, 1201, 1203, 1205, 1207, 1209, 1211, 1213, 1215, 1217, 1219, 1221, 1223, 1225, 1227, 1229, 1231, 1233, 1235, 1237, 1239, 1241, 1243, RNAi oligonucleotides for reducing or inhibiting PNPLA3 expression, which include antisense strands having 1245, 1247, 1249, 1251, 1253, 1255, 1257, 1259, 1261, 1263, 1265, 1267, 1269, 1271, 1273, 1275, 1277, 1279, 1281, 1283, 1285, 1287, 1289, 1291, 1293, 1295, 1297, 1299, or 1301), are described.

[0110] In some embodiments, RNAi oligonucleotides for reducing or inhibiting PNPLA3 expression, comprising an antisense strand and a sense strand, wherein the antisense strand has a sequence described in Table A, Table B, Table C, or Table D, and the sense strand has a sequence described in Table A, Table B, Table C, or Table D, are described.

[0111] In some examples, RNAi oligonucleotides are nucleotide sequences listed in Table A, Table B, Table C, or Table D, particularly SEQ ID NOs: 1178, 1180, 1182, 1184, 1186, 1188, 1190, 1192, 1194, 1196, 1198, 1200, 1202, 1204, 1206, 1208, 1210, 1212, 1214, 1216, 1218, 1220, 1222, 1224, 1226, 1228, 1230, 12 Includes a sense chain having one of the following: 32, 1234, 1236, 1238, 1240, 1242, 1244, 1246, 1248, 1250, 1252, 1254, 1256, 1258, 1260, 1262, 1264, 1266, 1268, 1270, 1272, 1274, 1276, 1278, 1280, 1282, 1284, 1286, 1288, 1290, 1292, 1294, 1296, 1298, or 1300. In some examples, RNAi oligonucleotides are nucleotide sequences listed in Tables A, B, C, or D, particularly SEQ ID NOs: 1179, 1181, 1183, 1185, 1187, 1189, 1191, 1193, 1195, 1197, 1199, 1202, 1203, 1205, 1207, 1209, 1211, 1213, 1215, 1217, 1219, 1221, 1223, 1225, 1227, 1229, 1231, 123 The antisense chain includes one of the following: 3, 1235, 1237, 1239, 1241, 1243, 1245, 1247, 1249, 1251, 1253, 1255, 1257, 1259, 1261, 1263, 1265, 1267, 1269, 1271, 1273, 1275, 1277, 1279, 1281, 1283, 1285, 1287, 1289, 1291, 1293, 1295, 1297, 1299, or 1301.In a specific example, RNAi oligonucleotides are sequence numbers 1178, 1180, 1182, 1184, 1186, 1188, 1190, 1192, 1194, 1196, 1198, 1200, 1202, 1204, 1206, 1208, 1210, 1212, 1214, 1216, 1218, 1220, 1222, 1224, 1226, 1228, 1230, 1232, 1234, 1236, 12 It has one of the following nucleotide sequences: 38, 1240, 1242, 1244, 1246, 1248, 1250, 1252, 1254, 1256, 1258, 1260, 1262, 1264, 1266, 1268, 1270, 1272, 1274, 1276, 1278, 1280, 1282, 1284, 1286, 1288, 1290, 1292, 1294, 1296, 1298, or 1300. The sense chain and sequence numbers 1179, 1181, 1183, 1185, 1187, 1189, 1191, 1193, 1195, 1197, 1199, 1201, 1203, 1205, 1207, 1209, 1211, 1213, 1215, 1217, 1219, 1221, 1223, 1225, 1227, 1229, 1231, 1233, 1235, 1237, 1239, 1241, 1243, 1 It comprises an antisense strand having one of the following nucleotide sequences: 245, 1247, 1249, 1251, 1253, 1255, 1257, 1259, 1261, 1263, 1265, 1267, 1269, 1271, 1273, 1275, 1277, 1279, 1281, 1283, 1285, 1287, 1289, 1291, 1293, 1295, 1297, 1299, or 1301.

[0112] In a specific example, the RNAi oligonucleotide is one of the following nucleotide sequences: 1188, 1190, 1200, 1216, 1218, 1220, 1224, 1230, 1232, 1234, 1244, 1246, 1250, 1254, 1262, 1288, 1290, 1292, 1294, 1296, 1298, or 1300. It comprises a sense strand having a column and an antisense strand having one of the following nucleotide sequences: SEQ ID NOs: 1189, 1191, 1201, 1215, 1217, 1221, 1225, 1231, 1233, 1235, 1245, 1247, 1251, 1255, 1263, 1289, 1291, 1295, 1297, 1299, or 1301.

[0113] In certain examples, an RNAi oligonucleotide comprises a sense strand having one nucleotide sequence of sequence numbers 1188, 1190, 1220, 1224, 1230, 1232, 1244, 1246, 1250, or 1254, and an antisense strand having one nucleotide sequence of sequence numbers 1189, 1191, 1221, 1225, 1231, 1233, 1245, 1247, 1251, or 1255.

[0114] In a specific example, the sense and antisense strands of the RNAi oligonucleotide are selected from the following: (a) Sequence IDs 1220 and 1221, (b) Sequence IDs 1224 and 1225, (c) Sequence IDs 1230 and 1231, (d) Sequence IDs 1232 and 1233, (e) Sequence IDs 1188 and 1189, (f) Sequence IDs 1190 and 1191, (g) Sequence IDs 1244 and 1245, (h) Sequence IDs 1250 and 1251-, (i) Sequence IDs 1254 and 1255, and (j) Sequence IDs 1246 and 1247.

[0115] Those skilled in the art will understand that in some examples, the sequences shown in sequence listings are referenced when describing the structure of oligonucleotides (e.g., ds oligonucleotides) or other nucleic acids. In such examples, the actual oligonucleotides or other nucleic acids have one or more alternative nucleotides (e.g., RNA counterparts of DNA nucleotides or DNA counterparts of RNA nucleotides) and / or one or more modified nucleotides and / or one or more modified nucleotide interbonds and / or one or more other modifications compared to the described sequence, while retaining essentially the same or similar complementary properties as the described sequence.

[0116] In some embodiments, the ds oligonucleotides herein comprise a 25-nucleotide sense strand and a 27-nucleotide antisense strand that, when acted upon by the Dicer enzyme, produces an antisense strand incorporated into the mature RISC. In certain examples, the sense strand of the ds oligonucleotide is longer than 27 nucleotides (e.g., 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, or 40 nucleotides). In certain examples, the sense strand of the ds oligonucleotide is longer than 25 nucleotides (e.g., 26, 27, 28, 29, or 30 nucleotides).

[0117] In some examples, one 5' end of the ds oligonucleotides herein is thermodynamically unstable compared to the other 5' end. In some embodiments, the ds oligonucleotides are asymmetric, with a blunt end at the 3' end of the sense strand and a 3' overhang at the 3' end of the antisense strand. In some embodiments, the 3' overhang on the antisense strand is about 1 to about 8 nucleotides long (e.g., 1, 2, 3, 4, 5, 6, 7, or 8 nucleotides). Typically, the ds oligonucleotides of RNAi have a 2-nucleotide overhang at the 3' end of the antisense (guide) strand. However, other overhangs are also possible. In some embodiments, the overhang is about 1 to about 6 nucleotides, optionally 1 to 5, 1 to 4, 1 to 3, 1 to 2, 2 to 6, 2 to 5, 2 to 4, 2 to 3, 3 to 6, 3 to 5, 3 to 4, 4 to 6, 4 to 5, 5 to 6 nucleotides, or a 3' overhang having a length of 1, 2, 3, 4, 5, or 6 nucleotides. However, in other examples, the overhang is approximately 1 to approximately 6 nucleotides, optionally 1 to 5, 1 to 4, 1 to 3, 1 to 2, 2 to 6, 2 to 5, 2 to 4, 2 to 3, 3 to 6, 3 to 5, 3 to 4, 4 to 6, 4 to 5, 5 to 6 nucleotides, or a 5' overhang containing a length of 1, 2, 3, 4, 5, or 6 nucleotides.

[0118] In some cases, the two terminal nucleotides at the 3' end of the antisense strand are modified. In some cases, the two terminal nucleotides at the 3' end of the antisense strand are complementary to the target mRNA (e.g., PNPLA3 mRNA). In other cases, the two terminal nucleotides at the 3' end of the antisense strand are not complementary to the target mRNA. In some cases, the two terminal nucleotides at each 3' end of the oligonucleotide in a nicked tetraloop structure are GG. Typically, one or both of the two terminal GG nucleotides at each 3' end of the ds oligonucleotide are not complementary to the target mRNA.

[0119] In some cases, there are one or more mismatches (e.g., one, two, three, four, or five) between the sense strand and the antisense strand. If there are two or more mismatches between the sense strand and the antisense strand, they may be contiguous (e.g., two, three, or more consecutively) or scattered throughout the complementarity region. In some cases, the 3' end of the sense strand contains one or more mismatches. In certain cases, two mismatches are incorporated into the 3' end of the sense strand. In some cases, mismatches or segmental destabilization at the 3' end of the sense strand of an oligonucleotide improve or increase the potency of the ds oligonucleotide.

[0120] A. Antisense strands: The oligonucleotides (e.g., ds oligonucleotides) herein for targeting PNPLA3 mRNA and inhibiting PNPLA3 expression include antisense strands containing sequences listed in Table 1 or Table 3, or Tables A, B, C, or D. In some examples, oligonucleotides include an antisense chain having at least about 12 (e.g., at least 12, at least 13, at least 14, at least 15, at least 16, at least 17, at least 18, at least 19, at least 20, at least 21, at least 22, or at least 23) consecutive nucleotides of the sequence described in any one of SEQ ID NOs. 788, 844, 868, 872, 938, 1004, 1008, 1018, 1162, or 1164, or an antisense chain having any one nucleotide sequence of SEQ ID NOs. 1189, 1191, 1221, 1225, 1231, 1233, 1245, 1247, 1251, or 1255.

[0121] Furthermore, the oligonucleotides herein (e.g., ds oligonucleotides) may include an antisense chain of up to approximately 40 nucleotides in length (e.g., up to 40, up to 35, up to 30, up to 27, up to 25, up to 21, up to 19, up to 17, or up to 12 nucleotides). In some examples, the oligonucleotides may have an antisense chain of at least approximately 12 nucleotides in length (e.g., at least 12, at least 15, at least 19, at least 21, at least 22, at least 25, at least 27, at least 30, at least 35, or at least 38 nucleotides). Alternatively, oligonucleotides may have antisense chains with a nucleotide length in the range of approximately 12 to approximately 40 (e.g., 12 to 40, 12 to 36, 12 to 32, 12 to 28, 15 to 40, 15 to 36, 15 to 32, 15 to 28, 17 to 22, 17 to 25, 19 to 27, 19 to 30, 20 to 40, 22 to 40, 25 to 40, or 32 to 40). In specific examples, oligonucleotides may have antisense chains with a nucleotide length of 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, or 40 nucleotides.

[0122] As described above, the antisense strands of oligonucleotides in this specification may sometimes be referred to as "guide strands." For example, an antisense strand incorporated into RISC and bound to an Argonaut protein, such as Ago2, or an antisense strand incorporated into or bound to one or more similar factors to induce silencing of a target gene, is called a guide strand (or passenger strand).

[0123] B. Sense strand: The oligonucleotides herein (e.g., ds oligonucleotides) for targeting PNPLA3 mRNA and inhibiting PNPLA3 expression include a sense strand sequence containing a sequence described in Table 1 or Table 3, or in Tables A, B, C, or D. In some examples, the oligonucleotides include a sense strand having at least about 12 (e.g., at least 13, at least 14, at least 15, at least 16, at least 17, at least 18, at least 19, at least 20, at least 21, at least 22, or at least 23) consecutive nucleotides of a sequence described in any one of SEQ ID NOs. 787, 843, 867, 871, 937, 1003, 1007, 1017, 1161, or 1163.

[0124] Furthermore, the oligonucleotides herein (e.g., ds oligonucleotides) include a sense strand (or passenger strand) of up to approximately 40 nucleotides in length (e.g., up to 40, up to 36, up to 30, up to 27, up to 25, up to 21, up to 19, up to 17, or up to 12 nucleotides). In some examples, the oligonucleotide may have a sense strand of at least approximately 12 nucleotides in length (e.g., at least 12, at least 15, at least 19, at least 21, at least 25, at least 27, at least 30, at least 36, or at least 38 nucleotides). Alternatively, oligonucleotides may have sense strands with a nucleotide length in the range of approximately 12 to approximately 40 (e.g., 12-40, 12-36, 12-32, 12-28, 15-40, 15-36, 15-32, 15-28, 17-21, 17-25, 19-27, 19-30, 20-40, 22-40, 25-40, or 32-40). In specific examples, oligonucleotides may have sense strands with a nucleotide length of 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, or 40 nucleotides.

[0125] In some embodiments, the sense strand includes a stem-loop structure at its 3' end. In other embodiments, the sense strand includes a stem-loop structure at its 5' end. In additional embodiments, the stem is a double helix approximately 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, or 14 bp long. In some embodiments, the stem-loop protects the oligonucleotide from degradation (e.g., enzymatic degradation) and facilitates or improves targeting and / or delivery to target cells, tissues, or organs (e.g., the liver), or both. For example, the loop of the stem-loop provides a nucleotide having one or more modifications that facilitate, improve, or increase targeting to target mRNA (e.g., PNPLA3 mRNA), inhibition of target gene expression (e.g., PNPLA3 expression), and / or delivery to target cells, tissues, or organs (e.g., the liver), or both. In some embodiments, the stem-loop itself or modifications to the stem-loop do not substantially affect the intrinsic gene expression inhibitory activity of the oligonucleotide, but they enhance stability (e.g., provide protection against degradation) and / or facilitate, improve, or increase the delivery of the oligonucleotide to target cells, tissues, or organs (e.g., the liver). In certain embodiments, the oligonucleotide includes a sense strand containing a stem-loop represented as S1-L-S2 (e.g., at its 3' end) (where S1 is complementary to S2, and L forms a single-stranded loop between S1 and S2 of up to approximately 10 nucleotides in length (e.g., 3, 4, 5, 6, 7, 8, 9, or 10 nucleotides in length). In certain embodiments, the loop (L) is 4 nucleotides in length. Figures 1 and 2 depict non-limiting examples of such oligonucleotides. In some embodiments, the loop (L) of the stem-loop having the above-described structure S1-L-S2 is a tetraloop (e.g., within a nicked tetraloop structure). In some embodiments, the tetraloop includes ribonucleotides, deoxyribonucleotides, modified nucleotides, delivery ligands, and combinations thereof.

[0126] V. Modification of oligonucleotides

[0127] A. Sugar Modification: Modified sugars (also called sugar analogs herein) include, for example, modified deoxyribose or ribose moieties in which one or more modifications occur at the 2', 3', 4', and / or 5' carbon positions of the sugar. Modified sugars may also include unnatural alternative carbon structures, such as those found in locked nucleic acids ("LNA"; see, e.g., Koshkin et al. (1998) Tetrahedon 54:3607-3630), unlocked nucleic acids ("UNA"; see, e.g., Snead et al. (2013) Mol.Ther-Nuc.Acids 2:e103), and cross-linked nucleic acids ("BNA"; see, e.g., Imanishi & Obika (2002) Chem.Commun. 16:1653-1659).

[0128] In some embodiments, the nucleotide modification in the sugar is a 2'-modification, such as 2'-O-propargyl, 2'-O-propylamine, 2'-amino, 2'-ethyl, 2'-fluoro(2'-F), 2'-aminoethyl(EA), 2'-O-methyl(2'-OMe), 2'-O-methoxyethyl(2'-MOE), 2'-O-[2-(methylamino)-2-oxoethyl](2'-O-NMA), or 2'-deoxy-2'-fluoro-β-d-arabinonucleotide(2'-FANA). In certain embodiments, the modification is 2'-F, 2'-OMe, or 2'-MOE. In other embodiments, the modification in the sugar is a modification of the sugar ring, and includes modification of one or more carbons of the sugar ring. For example, modifications in sugars include a 2'-oxygen atom of the sugar bonded to the 1'-carbon or 4'-carbon, or a 2'-oxygen atom bonded to the 1'-carbon or 4'-carbon via an ethylene or methylene bridge. In other embodiments, the modification is an acyclic sugar lacking a bond between the 2'-carbon and 3'-carbon. In other embodiments, the modification is a thiol group, such as a thiol group at the 4' position of the sugar.

[0129] The oligonucleotides herein contain at least one modified nucleotide (e.g., at least one, at least five, at least ten, at least fifteen, at least twenty, at least twenty-five, at least thirty, at least thirty-five, at least forty, at least forty-five, at least fifty, at least fifty, at least sixty, or more). In some embodiments, the sense strand contains at least one modified nucleotide (e.g., at least one, at least five, at least ten, at least fifteen, at least twenty, at least twenty-five, at least thirty, at least thirty-five, or more). In some embodiments, the antisense strand contains at least one modified nucleotide (e.g., at least one, at least five, at least ten, at least fifteen, at least twenty, or more).

[0130] In certain embodiments, all nucleotides of the sense strand are modified. Similarly, all nucleotides of the antisense strand are modified. In some embodiments, all nucleotides of the oligonucleotides herein (i.e., both the sense strand and the antisense strand) are modified. As described above, in some embodiments, the modified nucleotides are 2'-modified (e.g., 2'-F, 2'-OMe, 2'-MOE, and / or 2'-deoxy-2'-fluoro-β-d-arabinonucleotides). In certain embodiments, the modified nucleotides are 2'-modified, e.g., 2'-F or 2'-OMe.

[0131] Furthermore, the oligonucleotides described herein have different modification patterns. In some embodiments, the modified oligonucleotide comprises an antisense chain having a modification pattern described in any one of Tables A, B, C, or D, and a sense chain sequence having a modification pattern described in any one of Tables A, B, C, or D (and Figure 1). In some embodiments, one or more of the 8, 9, 10, or 11 positions of the sense chain are modified with 2'-F. In other embodiments, the sugar moieties of the nucleotides at positions 1-7 and 12-20 of the sense chain are modified with 2'-OMe.

[0132] In some embodiments, the antisense chain contains three nucleotides modified with 2'-F at the 2' position of the sugar moiety. In some embodiments, the sugar moieties at positions 2, 5, and 14 of the antisense chain, and optionally up to three nucleotides at positions 1, 3, 7, and 10, are modified with 2'-F. In other embodiments, the sugar moieties at positions 2, 5, and 14 of the antisense chain are modified with 2'-F. In yet another embodiment, the sugar moieties at positions 1, 2, 5, and 14 of the antisense chain are modified with 2'-F. In yet another embodiment, the sugar moieties at positions 1, 2, 3, 5, 10, and 14 of the antisense chain are modified with 2'-F. In yet another embodiment, the sugar moieties at positions 2, 3, 5, 7, 10, and 14 of the antisense chain are modified with 2'-F.

[0133] B. 5'-terminal phosphate: A 5'-terminal phosphate group can be used to facilitate the interaction between the oligonucleotides described herein and Ago2. However, oligonucleotides having a 5'-phosphate group are susceptible to degradation by phosphatases or other enzymes, which may limit their bioavailability in vivo. In some embodiments, the oligonucleotide (e.g., ds oligonucleotide) includes a 5'-phosphate analog that is resistant to such degradation. Examples of such phosphate analogs include, but are not limited to, oxymethylphosphonates, vinylphosphonates, malonylphosphonates, or combinations thereof. In certain embodiments, the 3' end of the oligonucleotide chain is bonded to a chemical moiety ("phosphate mimic") that mimics the electrostatic and steric properties of a natural 5'-phosphate group.

[0134] Alternatively, or further, the oligonucleotides herein have a phosphate analog at the 4' position of the sugar (referred to as the "4'-phosphate analog"). See, for example, International Patent Application Publication WO2018 / 045317. In some embodiments, the oligonucleotides herein include a 4'-phosphate analog at the 5' terminal nucleotide. In some embodiments, the phosphate analog is an oxymethylphosphonate or an analog in which the oxygen atom of the oxymethyl group is bonded to the sugar moiety (e.g., at its 4'-carbon). In other embodiments, the 4'-phosphate analog is a thiomethylphosphonate or aminomethylphosphonate or an analog in which the sulfur atom of the thiomethyl group or the nitrogen atom of the aminomethyl group is bonded to the 4'-carbon of the sugar moiety. In certain embodiments, the 4'-phosphate analog is an oxymethylphosphonate represented by the formula -O-CH2-PO(OH)2 or -O-CH2-PO(OR)2, where R is independently selected from H, CH3, an alkyl group, CH2CH2CN, CH2OCOC(CH3)3, CH2OCH2CH2Si(CH3)3, or a protecting group. In certain embodiments, the alkyl group is CH2CH3. In certain other embodiments, R is independently selected from H, CH3, or CH2CH3.

[0135] C. Modified Nucleoside Bonds: In addition to the modifications described above, the oligonucleotides herein include modified nucleoside bonds. In some examples, modification or substitution of the phosphate results in an oligonucleotide containing at least one (e.g., at least one, at least two, at least three, or at least five) modified nucleotide bonds. In some embodiments, the oligonucleotide contains about 1 to about 10 (e.g., 1 to 10, 2 to 8, 4 to 6, 3 to 10, 5 to 10, 1 to 5, 1 to 3, or 1 to 2) modified nucleotide bonds. In certain additional embodiments, the oligonucleotide contains 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 modified nucleotide bonds.

[0136] Examples of modified nucleotide interlinking bonds include, but are not limited to, phosphorodithioate bonds, phosphorothioate bonds, phosphotriester bonds, thionoalkylphosphonate bonds, thionalkylphosphotriester bonds, phosphoramidite bonds, phosphonate bonds, or boranophosphate bonds. In some embodiments, at least one modified nucleotide interlinking bond of any one of the oligonucleotides disclosed herein is a phosphorothioate bond.

[0137] In some embodiments, the oligonucleotides herein include a phosphorothioate bond between one or more of the following: positions 1 and 2 of the sense chain, positions 1 and 2 of the antisense chain, positions 2 and 3 of the antisense chain, positions 3 and 4 of the antisense chain, positions 20 and 21 of the antisense chain, and positions 21 and 22 of the antisense chain. In other embodiments, the oligonucleotides include a phosphorothioate bond between each of the following: positions 1 and 2 of the sense chain, positions 1 and 2 of the antisense chain, positions 2 and 3 of the antisense chain, positions 20 and 21 of the antisense chain, and positions 21 and 22 of the antisense chain.

[0138] D. Base Modification: In addition to the modifications described above, the oligonucleotides herein also contain one or more modified nucleic acid bases. In some embodiments, the modified nucleic acid base (also referred to herein as a base analog) is attached to the 1' position of the nucleotide sugar moiety. In certain embodiments, the modified nucleic acid base is a nitrogen base. In certain other embodiments, the modified nucleic acid base does not contain a nitrogen atom. See, for example, U.S. Patent Application Publication 2008 / 0274462. In some embodiments, the modified nucleotide is a universal base. However, in certain embodiments, the modified nucleotide does not contain a nucleic acid base (base-free).

[0139] Regarding universal bases, universal bases include heterocyclic moieties located at the 1' position of the nucleotide sugar moiety of a modified nucleotide, or at the equivalent position of a nucleotide sugar moiety substitution, and when present in a double helix, they are positioned to pair with multiple types of bases without substantially altering the structure of the double helix. Furthermore, compared to a reference ss nucleic acid (e.g., oligonucleotide) that is perfectly complementary to the target nucleic acid, ss nucleic acids with universal bases have a lower T than the double helix formed with the complementary nucleic acid. m It forms a double helix with the target nucleic acid which has the universal base. However, compared to the reference ss nucleic acid in which the universal base is replaced by a base, resulting in one mismatch, the ss nucleic acid with the universal base has a higher T than the double helix formed with the nucleic acid with the mismatch base. m It forms a double helix with the target nucleic acid which has the following properties.

[0140] Examples of universally bound nucleotides include, but are not limited to, inosine, 1-β-D-ribofuranosyl-5-nitroindole, and / or 1-β-D-ribofuranosyl-3-nitropyrrole (see, e.g., U.S. Patent Application Publication No. 2007 / 0254362; Van Aerschot et al. (1995) Nucleic Acids Res. 23:4363-70; Loakes et al. (1995) Nucleic Acids Res. 23:2361-66; and Loakes & Brown (1994) Nucleic Acids Res. 22:4039-43).

[0141] E. Reversible Modifications: Certain modifications can be made to protect the oligonucleotides described herein from the in vivo environment before they reach target cells, but these modifications may reduce the potency or activity of the oligonucleotides once they reach the cytosol of the target cells. Therefore, reversible modifications can be made such that the molecules retain the desired properties outside the cell and are subsequently removed once they enter the cytosolic environment of the cell. Reversible modifications can be removed, for example, by the action of intracellular enzymes or by intracellular chemical conditions (e.g., through reduction by intracellular glutathione).

[0142] In some embodiments, reversibly modified nucleotides include a glutathione-sensitive moiety. Typically, nucleic acid molecules are chemically modified with a cyclic disulfide moiety to shield the negative charge generated by internucleotide diphosphate bonds, thereby improving cellular uptake and nuclease resistance. See U.S. Patent Application Publication 2011 / 0294869, International Patent Application Publications WO2014 / 088920 and WO2015 / 188197, and Meade et al. (2014) Nat. Biotechnol. 32:1256-1263. This reversibly modified internucleotide diphosphate bond is designed to be cleaved intracellularly by the reducing environment of the cytosol (e.g., glutathione). An earlier example is neutralizing phosphotriester modification, which has been reported to be cleavable inside cells (see, for example, Dellinger et al. (2003) J.Am.Chem.Soc.125:940-50).

[0143] Such reversible modifications protect the oligonucleotide during in vivo administration (e.g., through blood and / or cellular lysosome / endosomal compartments) where it will be exposed to nucleases and other harsh environmental conditions (e.g., pH). When released into the cellular cytosol, where glutathione levels are higher than in the extracellular space, the modifications are reversed, resulting in the cleavage of the oligonucleotide. Using a reversible glutathione-sensitive moiety makes it possible to introduce sterically larger chemical groups into the oligonucleotide compared to options obtained using irreversible chemical modifications. This means that these larger chemical groups will be removed in the cytosol, and therefore the biological activity of the oligonucleotide should not be interfered with within the cellular cytosol. As a result, these larger chemical groups can be manipulated to impart various advantages to the oligonucleotide, such as nuclease resistance, lipophilicity, charge, thermal stability, specificity, and reduced immunogenicity. In some examples, the structure of the glutathione-sensitive moiety can be manipulated to alter the dynamics of its release.

[0144] In some embodiments, the glutathione-sensitive moiety is bound to the sugar of the nucleotide. In certain embodiments, the glutathione-sensitive moiety is bound to the 2'-carbon of the sugar of the modified nucleotide. Additionally or alternatively, the glutathione-sensitive moiety is bound to the 5'-carbon of the sugar, particularly when the modified nucleotide is the 5'-terminal nucleotide of an oligonucleotide. Additionally or alternatively, the glutathione-sensitive moiety is bound to the 3'-carbon of the sugar, particularly when the modified nucleotide is the 3'-terminal nucleotide of an oligonucleotide. In some embodiments, the glutathione-sensitive moiety includes a sulfonyl group (see, for example, International Patent Application Publication WO2018 / 039364).

[0145] VI. Targeted Ligands

[0146] It is desirable to target one or more cells or one or more organs with the oligonucleotides described herein. Such a strategy may help to avoid undesirable effects in other organs or to avoid excessive loss of oligonucleotides to cells, tissues, or organs that do not benefit from them. Therefore, oligonucleotides may be modified to facilitate targeting and / or delivery to tissues, cells, or organs (e.g., to facilitate delivery of oligonucleotides to the liver). In some embodiments, oligonucleotides are modified to facilitate delivery of oligonucleotides to hepatocytes in the liver. In certain embodiments, the oligonucleotide comprises at least one nucleotide (e.g., one, two, three, four, five, six, or more nucleotides) conjugated to one or more targeting ligands.

[0147] Examples of targeting ligands include, but are not limited to, carbohydrates, amino sugars, cholesterol, peptides, polypeptides, proteins or parts of proteins (e.g., antibodies or antibody fragments), or lipids. In some embodiments, the targeting ligand is an aptamer. For example, the targeting ligand is an RGD peptide for targeting tumor vascular systems or glioma cells, a CREKA peptide for targeting tumor vascular systems or tumor stroma, transferrin, lactoferrin, or an aptamer for targeting transferrin receptors expressed on CNS vascular systems, or an anti-EGFR antibody for targeting EGFR on glioma cells. In certain embodiments, the targeting ligand is one or more GalNAc moieties.

[0148] In some embodiments, one or more nucleotides (e.g., one, two, three, four, five, or six) of an oligonucleotide can be conjugated to distinct targeting ligands. In some embodiments, two to four nucleotides of an oligonucleotide are conjugated to distinct targeting ligands. In some embodiments, the targeting ligand can be conjugated to two to four nucleotides at the end of either the sense or antisense strand (e.g., the targeting ligand is conjugated to the overhang or extension of two to four nucleotides at the 5' or 3' end of the sense or antisense strand), resulting in the targeting ligand resembling the bristles of a toothbrush and the oligonucleotide resembling a toothbrush. For example, an oligonucleotide may contain a stem loop at either the 5' or 3' end of the sense strand, and one, two, three, or four nucleotides of the stem loop can be individually conjugated to targeting ligands. In some embodiments, the oligonucleotide (e.g., ds oligonucleotide) includes a stem loop at the 3' end of the sense strand, the loop of the stem loop includes a triloop or tetraloop, and the three or four nucleotides of the triloop or tetraloop are each individually conjugated to a targeting ligand.

[0149] GalNAc is a high-affinity ligand for ASGPR, which is primarily expressed on the sinusoidal surface of hepatocytes, and plays a major role in the binding, internalization, and subsequent efflux of circulating glycoproteins (asialoglycoproteins) containing terminal galactose or GalNAc residues. The conjugation (either indirectly or directly) of a GalNAc moiety to an oligonucleotide as described herein is used to target the oligonucleotide to ASGPR expressed on a cell. In some embodiments, the oligonucleotide is conjugated to at least one GalNAc moiety, which causes the oligonucleotide to target ASGPR expressed on human liver cells (e.g., human hepatocytes).

[0150] Oligonucleotides are conjugated directly or indirectly to monovalent GalNAc. In some embodiments, oligonucleotides are conjugated directly or indirectly to two or more monovalent GalNAc (i.e., conjugated to two, three, or four monovalent GalNAc moieties, typically three or four monovalent GalNAc moieties). In some embodiments, oligonucleotides are conjugated to one or more divalent, trivalent, or tetravalent GalNAc moieties.

[0151] In some embodiments, one or more nucleotides (e.g., one, two, three, four, five, or six) of the oligonucleotides herein may be conjugated to a GalNAc moiety. In some embodiments, two to four nucleotides of a tetraloop are each conjugated to a separate GalNAc. In other embodiments, one to three nucleotides of a triloop are each conjugated to a separate GalNAc. In some embodiments, the targeting ligand is conjugated to two to four nucleotides at the end of either the sense or antisense strand (e.g., the ligand is conjugated to the overhang or extension of two to four nucleotides at the 5' or 3' end of the sense or antisense strand), resulting in the GalNAc moiety resembling the bristles of a toothbrush and the oligonucleotide resembling a toothbrush. In some embodiments, the GalNAc moiety is conjugated to nucleotides of the sense strand. For example, four GalNAc moieties are conjugated to nucleotides within the tetraloop of the sense strand, with each GalNAc moiety being conjugated to one nucleotide. In a particular embodiment, three GalNAc moieties are conjugated to nucleotides within the tetraloop of the sense strand, with each GalNAc moiety being conjugated to one nucleotide.

[0152] In certain embodiments, the oligonucleotide comprises a monovalent GalNAc bonded to a guanine nucleotide, referred to as [ademG-GalNAc] or 2'-aminodiethoxymethanol-guanine-GalNAc, as shown below. [ka]

[0153] In certain embodiments, the oligonucleotides herein include monovalent GalNAc bound to an adenine nucleotide, referred to as [ademA-GalNAc] or 2'-aminodiethoxymethanol-adenine-GalNAc, as shown below. [ka]

[0154] An example of such a conjugation is shown below for a loop having the nucleotide sequence GAAA in the 5'-3' direction (L = linker, X = heteroatom). Stem junctions are indicated in the formula. Such loops are listed, for example, in Table 3, A, B, C, or D, and are located at positions 27-30 of the sense strand shown in Figure 1. In the chemical formula, [ka] This is used to represent the binding site to the oligonucleotide chain. [ka]

[0155] The targeted ligand is conjugated to the nucleotide using an appropriate method or chemical technique (e.g., click chemistry). One method for conjugating the targeted ligand to the nucleotide is by using a click linker. In some embodiments, an acetal-based linker is used to conjugate the targeted ligand to one nucleotide of any of the oligonucleotides herein. Acetal-based linkers are disclosed, for example, in International Patent Application Publication WO2016 / 100401. In some examples, the linker is an unstable linker. However, in other examples, the linker is stable. An example of a loop having nucleotide GAAA in the 5'-3' direction, where the GalNAc portion is conjugated to the nucleotide of the loop using an acetal linker, is shown below. Such a loop is located, for example, at positions 27-30 of any one of the sense strands listed in Table 3, A, B, C, or D. In the chemical formula, [ka] This is a binding site to the oligonucleotide chain. [ka]

[0156] In some embodiments, a double-chain extension (e.g., up to 3, 4, 5, or 6 bp in length) is provided between the targeting ligand (e.g., a GalNAc portion) and the oligonucleotide of this specification (e.g., a ds oligonucleotide). In other embodiments, the oligonucleotide does not have GalNAc conjugated thereto.

[0157] Pharmaceutical preparations and pharmaceutical compositions

[0158] Oligonucleotides as used herein are incorporated into formulations or pharmaceutical compositions. Various formulations have been developed to facilitate the use of oligonucleotides. For example, oligonucleotides can be delivered to a target or cellular environment using formulations that minimize degradation, promote delivery and / or uptake, or impart other beneficial properties to the oligonucleotides in the formulation. In some embodiments, oligonucleotides are formulated in buffers such as phosphate-buffered saline, liposomes, micelle structures, and capsids.

[0159] Formulations of oligonucleotides containing cationic lipids are used to promote the transfection of oligonucleotides into cells. Cationic lipids such as lipofectin, cationic glycerol derivatives, and polycationic molecules (e.g., polylysine) can be used. Suitable lipids include oligofectamine, lipofectamine (Life Technologies), NC388 (Ribozyme Pharmaceuticals, Inc.), or FuGene6 (Roche), all of which should be used according to the manufacturer's instructions.

[0160] Therefore, in some embodiments, the formulations herein may comprise liposomes, lipids, lipid complexes, microspheres, microparticles, nanospheres, or nanoparticles (e.g., lipid nanoparticles), or may be formulated in other forms for administration to cells, tissues, organs, or bodies of an individual requiring administration (e.g., Remington, “The Science and Practice of Pharmacy” (LV Allen Jr., ed., 22 nd (See Edition, Pharmaceutical Press, 2013).

[0161] In some embodiments, the formulations herein further include excipients that can impart to the composition improved stability, improved absorption, improved solubility, and / or therapeutic enhancement effects of the active ingredient. In some embodiments, the excipient is a buffer (e.g., sodium citrate, sodium phosphate, Tris base, or sodium hydroxide) or a vehicle (e.g., buffer solution, petrolatum, dimethyl sulfoxide, or mineral oil). In some embodiments, the oligonucleotides herein are lyophilized to extend their shelf life and then put into solution prior to use (e.g., administration to an individual). Therefore, the excipient in a pharmaceutical composition containing one or more oligonucleotides is a lyophilization protectant (e.g., mannitol, lactose, polyethylene glycol, or polyvinylpyrrolidone) or a decay temperature modifier (e.g., dextran, Ficoll®, or gelatin).

[0162] Pharmaceutical compositions are formulated to suit their intended route of administration. Routes of administration include, but are not limited to, parenteral (e.g., intravenous, intramuscular, intraperitoneal, intradermal, and subcutaneous), oral (e.g., inhalation), transdermal (e.g., topical), transmucosal, and rectal administration.

[0163] Pharmaceutical compositions suitable for injection include sterile aqueous solutions (if water-soluble) or dispersions and sterile powders for the immediate preparation of sterile injection solutions or dispersions. For intravenous administration, suitable carriers include, but are not limited to, physiological saline, bacteriostatic water, Cremophor EL® (BASF), or phosphate-buffered saline (PBS). The carrier is a solvent or dispersion medium containing, for example, water, ethanol, polyols (e.g., glycerol, propylene glycol, liquid polyethylene glycol, etc.), and suitable mixtures thereof. In many embodiments, it is preferable to include isotonic agents, such as sugars, polyhydric alcohols, such as mannitol, sorbitol, and / or sodium chloride in the composition. A sterile injection solution is prepared by adding the required amount of the oligonucleotides specified herein to a selected solvent, along with one or a combination of the components listed above as needed, and then sterilizing by filtration.

[0164] Furthermore, the pharmaceutical composition contains at least about 0.1% or more of the therapeutic agent (e.g., one or more of the oligonucleotides specified herein), although the percentage of the therapeutic agent may be about 1% to about 80% or more of the total weight or volume of the composition. Factors such as solubility, bioavailability, biological half-life, route of administration, shelf life of the product, and other pharmacological considerations are conceivable by those skilled in the art in the field of preparing such pharmaceutical formulations, and therefore, various dosages and treatment regimens may be desirable.

[0165] While some examples focus on the delivery of at least one liver target of the oligonucleotides described herein, targeting of other tissues is also intended.

[0166] kit

[0167] The oligonucleotides herein may be incorporated into a kit comprising one or more oligonucleotides herein and instructions for use. In some embodiments, the kit comprises one or more oligonucleotides and accompanying documentation comprising instructions for use of the kit and / or any of its components. In other embodiments, the kit comprises a suitable container, one or more oligonucleotides, one or more controls, and various buffers, reagents, enzymes, and other standard components known in the art.

[0168] In some embodiments, the container may be at least one vial, well, test tube, flask, bottle, syringe, or other container means that holds one or more oligonucleotides and, optionally, appropriately dispenses them. In other embodiments where additional components are provided, the kit includes additional containers in which these components are held. The kit also includes means for tightly sealing one or more oligonucleotides and any other reagents for commercial sale. Such containers include injection-molded or blow-molded plastic containers that hold the desired vials. The container and / or kit includes a label with instructions for use and / or warnings.

[0169] In some embodiments, the kit includes one or more oligonucleotides as herein and a pharmaceutically acceptable carrier, or a pharmaceutical composition comprising one or more oligonucleotides, and instructions for doing so in an individual requiring treatment or delay of progression of a disease, disorder, or condition related to PNPLA3 expression.

[0170] method

[0171] Manufacturing method

[0172] The oligonucleotides described herein are prepared using methods and / or techniques known to those skilled in the art, such as conventional solid-phase nucleic acid synthesis. The polynucleotides of the oligonucleotides are assembled on a suitable nucleic acid synthesizer using standard nucleotide or nucleoside precursors (e.g., phosphoramidites). Automated nucleic acid synthesizers, such as DNA / RNA synthesizers, are commercially available from companies such as Applied Biosystems (Foster City, CA), BioAutomation (Irving, TX), and GE Healthcare Life Sciences (Pittsburgh, PA).

[0173] As those skilled in the art will understand, other methods and / or techniques for synthesizing the oligonucleotides described herein are used. Furthermore, various synthetic steps are carried out in different orders or sequences to obtain the desired compounds. Other synthetic chemical transformations, protecting groups (e.g., hydroxyl, amino, etc., present on bases), and methodologies for protecting and deprotecting oligonucleotides that are useful in the synthesis of oligonucleotides are known in the art, e.g., Larock, “Comprehensive Organic Transformations,” VCH Publishers (1989); Greene & Wuts, Protective Groups in Organic Synthesis, 2 nd It is described in Ed., John Wiley & Sons (1991); Fieser & Fieser, Fieser and Fieser's Reagents for Organic Synthesis, John Wiley & Sons (1994); and Paquette, ed., Encyclopedia of Reagents for Organic Synthesis, John Wiley & Sons (1995).

[0174] How to use

[0175] I. Methods for reducing PNPLA3 expression in cells, tissues, organs, and organisms.

[0176] The oligonucleotides described herein are used to reduce PNPLA3 mRNA in cells, tissues, organs, or organisms. The methods comprise the steps described herein, which may be carried out in the order described, although this is not necessarily required. However, other orders are also possible. Furthermore, individual or multiple steps may be carried out in parallel and / or overlapping time and / or individually or multiple times. Additionally, the methods may include additional, unspecified steps.

[0177] The method involves contacting or delivering an effective amount of any of the oligonucleotides specified herein (e.g., ds oligonucleotides) to a cell, cell population, tissue, organ, or individual in order to reduce PNPLA3 expression. In some embodiments, the reduction in PNPLA3 expression is determined by measuring a decrease in the amount or level of PNPLA3 mRNA, PNPLA3 protein, or PNPLA3 activity in the cell.

[0178] Regarding appropriate cell types, cell types are any cells that express mRNA (e.g., hepatocytes, macrophages, monocyte-derived cells, prostate cancer cells, brain, endocrine tissue, bone marrow, lymph nodes, lungs, gallbladder, liver, duodenum, small intestine, pancreas, kidneys, gastrointestinal tract, bladder, adipose and soft tissue, and skin cells). In some embodiments, the cells are primary cells obtained from an organism. In some embodiments, the primary cells have undergone a limited number of passages so that the cells substantially retain their native phenotypic characteristics. In some embodiments, the cells are ex vivo, in vivo, or in vitro cells (i.e., such that one or more oligonucleotides herein can be delivered to cells in culture or to an organism in which the cells reside).

[0179] In some embodiments, the oligonucleotides herein are delivered to cells or cell populations using nucleic acid delivery methods known in the art, including but not limited to injection of a solution containing the oligonucleotide, bombarding with oligonucleotide-coated particles, exposure of cells or cell populations to a solution containing the oligonucleotide, or electroporation of cell membranes in the presence of the oligonucleotide. Other methods known in the art for delivering oligonucleotides to cells are used, for example, lipid-mediated transport, chemical-mediated transport, and cationic liposome transfection such as calcium phosphate.

[0180] The reduction in PNPLA3 expression is determined by an assay or technique that evaluates one or more molecules, properties, or characteristics of cells or cell populations associated with PNPLA3 expression (e.g., using a PNPLA3 expression biomarker), or by an assay or technique that evaluates a molecule that directly indicates PNPLA3 expression in cells or cell populations (e.g., PNPLA3 mRNA or PNPLA3 protein). In some embodiments, the extent to which an oligonucleotide reduces PNPLA3 expression is evaluated by comparing PNPLA3 expression in cells or cell populations contacted with the oligonucleotide to that of control cells or cell populations (e.g., cells or cell populations not contacted with the oligonucleotide or contacted with a control oligonucleotide). In some embodiments, a control amount or level of PNPLA3 expression in control cells or cell populations is predetermined so that it is not necessary to measure the control amount or level in all cases in which the assay or technique is performed. The predetermined level or value can take various forms, including, but not limited to, a single cutoff value such as a median or mean.

[0181] Contact with or delivery of the oligonucleotides of this specification (e.g., ds oligonucleotides) to cells or cell populations results in a decrease in PNPLA3 expression. In some embodiments, the decrease in PNPLA3 expression is compared to a control amount or level of PNPLA3 expression in cells or cell populations that have not been contacted with the oligonucleotide or that have been contacted with a control oligonucleotide. In some embodiments, the decrease in PNPLA3 expression is about ≤1%, ≤5%, ≤10%, ≤15%, ≤20%, ≤25%, ≤30%, ≤35%, ≤40%, ≤45%, ≤50%, ≤55%, ≤60%, ≤70%, ≤80%, or ≤90% compared to a control amount or level of PNPLA3 expression. In some embodiments, the control amount or level of PNPLA3 expression is the amount or level of PNPLA3 mRNA and / or PNPLA3 protein in cells or cell populations that have not been contacted with the oligonucleotides of this specification. In some embodiments, the effect of oligonucleotide delivery to cells or cell populations by the methods herein is evaluated after any finite period or a fixed time (e.g., minutes, hours, days, weeks, and / or months). For example, PNPLA3 expression is determined in cells or cell populations at least about 4 hours, about 8 hours, about 12 hours, about 18 hours, or about 24 hours. Alternatively, PNPLA3 expression is determined in cells or cell populations at least about 1 day, about 2 days, about 3 days, about 4 days, about 5 days, about 6 days, about 7 days, about 8 days, about 9 days, about 10 days, about 11 days, about 12 days, about 13 days, about 14 days, about 21 days, about 28 days, about 35 days, about 42 days, about 49 days, about 56 days, about 63 days, about 70 days, about 77 days, or about 84 days or longer after contacting or delivery of the oligonucleotide to the cells or cell population. In other embodiments, PNPLA3 expression is determined in cells or cell populations at least about 1 month, about 2 months, about 3 months, about 4 months, about 5 months, or about 6 months or longer after contacting or delivering oligonucleotides to the cells or cell populations.

[0182] In some embodiments, the oligonucleotides of this specification are delivered in the form of a transgene engineered to express one or more oligonucleotides or chains (e.g., sense and antisense strands) in a cell. For example, an oligonucleotide is delivered using a transgene engineered to express any of the oligonucleotides of this specification. The transgene may be delivered using a viral vector (e.g., adenovirus, retrovirus, vaccinia virus, poxvirus, adeno-associated virus, or herpes simplex virus) or a non-viral vector (e.g., plasmid or synthetic mRNA). In some embodiments, the transgene is injected directly into an organism.

[0183] II. Treatment method:

[0184] A method for treating an individual who has, is suspected of having, or is at risk of developing a disease, disorder, or condition related to PNPLA3 expression comprises administering at least one oligonucleotide of this specification. Furthermore, a method for treating or attenuating the onset or progression of a disease, disorder, or condition related to PNPLA3 expression in an individual comprises using one or more oligonucleotides of this specification. Furthermore, a method for achieving one or more therapeutic effects in a subject having a disease, disorder, or condition related to PNPLA3 expression comprises providing one or more oligonucleotides of this specification. In some embodiments, an individual can be treated by administering one or more therapeutically effective amounts of any one of the embodiments disclosed above. In some embodiments, the treatment comprises reducing PNPLA3 expression. In some embodiments, the individual is treated therapeutically. In some embodiments, the individual is treated prophylactically. In all of these embodiments, the oligonucleotide is selected from Tables A, B, C, or D.

[0185] In some embodiments, one or more oligonucleotides, or a pharmaceutical composition containing oligonucleotides, is administered to an individual having a disease, disorder, or condition related to PNPLA3 expression, resulting in a decrease in PNPLA3 expression in the individual, thereby treating the individual. In some embodiments, the amount or level of PNPLA3 mRNA is decreased in the individual. In other embodiments, the amount or level of PNPLA3 protein is decreased in the individual. In yet another embodiment, the amount or level of PNPLA3 activity is decreased in the individual. In yet another embodiment, the amount or level of hepatic TG (e.g., one or more TG or total TG in the liver) is decreased in the individual, particularly in the liver. In yet another example, the amount or level of hepatic inflammation may be decreased. In yet another example, the amount or level of hepatic fibrosis is decreased. In yet another embodiment, the amount or level of plasma AST, plasma ALT, or even Pro-C3 is decreased. In any of the embodiments disclosed above, the oligonucleotide comprises a sense strand having one nucleotide sequence of SEQ ID NOs: 1188, 1190, 1220, 1224, 1230, 1232, 1244, 1246, 1250, or 1254, and an antisense strand having one nucleotide sequence of SEQ ID NOs: 1189, 1191, 1221, 1225, 1231, 1233, 1245, 1247, 1251, or 1255.

[0186] In some embodiments, PNPLA3 expression in an individual is reduced by at least about 30%, about 35%, about 40%, about 45%, about 50%, about 55%, about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, about 90%, about 95%, about 99%, or more than 99% compared to PNPLA3 expression before administration of one or more oligonucleotides or their pharmaceutical composition. In other embodiments, PNPLA3 expression in an individual is reduced by at least about 30%, about 35%, about 40%, about 45%, about 50%, about 55%, about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, about 90%, about 95%, about 99%, or more than 99% compared to PNPLA3 expression in an individual that has not been administered one or more oligonucleotides or pharmaceutical compositions, or in an individual that has been treated with a control oligonucleotide, pharmaceutical composition, or a treatment (e.g., a reference or control subject).

[0187] In certain embodiments, the amount or level of PNPLA3 mRNA in an individual is reduced by at least about 30%, about 35%, about 40%, about 45%, about 50%, about 55%, about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, about 90%, about 95%, about 99%, or more than 99% compared to the amount or level of PNPLA3 mRNA before administration of one or more oligonucleotides or the pharmaceutical composition thereof. In some embodiments, the amount or level of PNPLA3 mRNA in an individual is reduced by at least about 30%, about 35%, about 40%, about 45%, about 50%, about 55%, about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, about 90%, about 95%, about 99%, or more than 99% compared to an individual that has not been administered one or more oligonucleotides or pharmaceutical compositions, or an individual that has been treated with a control oligonucleotide, pharmaceutical composition, or a reference or control subject.

[0188] In certain embodiments, the amount or level of PNPLA3 protein in an individual is reduced by at least about 30%, about 35%, about 40%, about 45%, about 50%, about 55%, about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, about 90%, about 95%, about 99%, or more than 99% compared to the amount or level of PNPLA3 protein before administration of one or more oligonucleotides or the pharmaceutical composition. In other embodiments, the amount or level of PNPLA3 protein in an individual is reduced by at least about 30%, about 35%, about 40%, about 45%, about 50%, about 55%, about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, about 90%, about 95%, about 99%, or more than 99% compared to an individual that has not been administered one or more oligonucleotides or pharmaceutical compositions, or an individual that has been treated with a control oligonucleotide, pharmaceutical composition, or a reference or control subject.

[0189] In certain embodiments, the amount or level of PNPLA3 activity in an individual is reduced by at least about 30%, about 35%, about 40%, about 45%, about 50%, about 55%, about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, about 90%, about 95%, about 99%, or more than 99% compared to the amount or level of PNPLA3 activity before administration of one or more oligonucleotides or the pharmaceutical composition. In some embodiments, the amount or level of PNPLA3 activity in an individual is reduced by at least about 30%, about 35%, about 40%, about 45%, about 50%, about 55%, about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, about 90%, about 95%, about 99%, or more than 99% compared to an individual that has not been administered one or more oligonucleotides or pharmaceutical compositions, or an individual that has been treated with a control oligonucleotide, pharmaceutical composition, or other treatment (e.g., a reference or control subject).

[0190] In certain embodiments, the amount or level of TG in an individual, particularly liver TG, may be reduced by at least about 30%, about 35%, about 40%, about 45%, about 50%, about 55%, about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, about 90%, about 95%, about 99%, or more than 99% compared to the amount or level of TG before administration of one or more oligonucleotides or their pharmaceutical composition. In some embodiments, the amount or level of TG in an individual is reduced by at least about 30%, about 35%, about 40%, about 45%, about 50%, about 55%, about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, about 90%, about 95%, about 99%, or more than 99% compared to the amount or level of TG in an individual that has not been administered one or more oligonucleotides or pharmaceutical compositions, or in an individual that has been treated with a control oligonucleotide, pharmaceutical composition, or a reference or control subject.

[0191] In this specification, the amount or level of PNPLA3 expression, PNPLA3 mRNA, PNPLA3 protein, PNPLA3 activity, liver TG, or any combination thereof is reduced in cells obtained from or isolated from an individual (e.g., hepatocytes), cell populations or groups (e.g., organoids), tissues (e.g., liver tissue), specimens (e.g., liver biopsy specimens), organs (e.g., liver), blood or fractions thereof (e.g., plasma), or any other biomaterial. In some embodiments, the amount or level of PNPLA3 expression, PNPLA3 mRNA, PNPLA3 protein, PNPLA3 activity, TG, or any combination thereof is reduced in two or more types of cells (e.g., hepatocytes and one or more other types of cells), two or more cell populations, two or more types of tissues (e.g., liver tissue and one or more other types of tissues), two or more samples (e.g., liver biopsy samples and one or more other types of biopsy samples), two or more organs (e.g., liver and one or more other organs), or two or more blood fractions (e.g., plasma and one or more other blood fractions).

[0192] Examples of diseases, disorders, or conditions associated with PNPLA3 expression include, but are not limited to, AH, ALD, HCC, CCA, PSC, NAFLD, NASH, fatty liver (steatosis), liver inflammation, liver fibrosis, cirrhosis, or combinations thereof.

[0193] Due to their high specificity, the oligonucleotides herein specifically target the mRNA of target genes in cells, tissues, or organs (e.g., the liver). In disease prevention, target genes are genes required for the onset or maintenance of the disease, or genes identified as being associated with a high risk of developing the disease. In disease treatment, one or more oligonucleotides are brought into contact with cells, tissues, or organs that exhibit or mediate the disease. For example, oligonucleotides substantially identical to all or part of a wild-type (i.e., native) or mutant gene associated with a disorder or condition related to PNPLA3 expression are brought into contact with or introduced into a target cell or tissue type, such as hepatocytes or other liver cells.

[0194] In some embodiments, the target gene is derived from any mammal, such as a human. Any gene can be silenced according to the methods herein. Furthermore, the methods herein typically involve administering to an individual a therapeutically effective dose, i.e., an amount of one or more oligonucleotides herein that can produce a desired therapeutic outcome. The therapeutically permissible dose is the amount that therapeutically treats a disease or disorder or condition. An appropriate dose for any single individual is determined depending on specific factors, including the individual's size, body surface area, age, the composition administered, the active ingredient(s) in the composition, the time and route of administration, overall health status, and other therapeutic agents being administered concurrently.

[0195] In this method, an individual is administered one of the oligonucleotides or compositions of this specification by enteral (e.g., orally, by a gastric feeding tube, by a duodenal feeding tube, via gastrostomy, or rectally), parenterally (e.g., subcutaneous injection, intravenous injection or infusion, intra-arterial injection or infusion, intramedullary infusion, intramuscular injection, intracerebral injection, intraventricular injection, or intrathecal cavity), topically (e.g., percutaneously, by inhalation, via eye drops, or through mucosa), or by direct injection into a target organ (e.g., the liver of the subject). Typically, oligonucleotides are administered intravenously or subcutaneously.

[0196] As a non-limiting set of examples, the oligonucleotides or compositions herein are typically administered quarterly (once every three months), bimonthly (once every two months), monthly, or weekly. For example, the oligonucleotides or compositions are administered weekly, or at intervals of two or three weeks. In certain embodiments, the oligonucleotides or compositions are administered daily. In some embodiments, an individual is administered one or more loading doses of the oligonucleotide or composition, followed by one or more maintenance doses of the oligonucleotide or composition.

[0197] In some embodiments, the individual is a human, a non-human primate, or other mammal. In other embodiments, the individual is a domestic animal such as a dog or cat, a livestock such as a horse, a cow, a pig, a sheep, a goat, or a chicken, and an animal such as a mouse, a rat, a guinea pig, or a hamster.

[0198] III. Medical Uses The oligonucleotides herein can be used or adapted for use to treat individuals who would benefit from reduced PNPLA3 expression (e.g., humans having diseases, disorders, or conditions related to PNPLA3 expression). In some embodiments, oligonucleotides are provided for use or adapted for use to treat individuals having diseases, disorders, or conditions related to PNPLA3 expression. Also provided are oligonucleotides for use or adaptable for use in the manufacture of pharmaceuticals or pharmaceutical compositions for treating diseases, disorders, or conditions related to PNPLA3 expression. In other embodiments, oligonucleotides are provided for use or adaptable for use to target PNPLA3 mRNA and reduce PNPLA3 expression (e.g., via the RNAi pathway). In other embodiments, oligonucleotides are provided for use or adaptable for use to target PNPLA3 mRNA and reduce the amount or level of PNPLA3 mRNA, PNPLA3 protein, and / or PNPLA3 activity.

[0199] In some embodiments, the method includes selecting individuals for treatment based on individuals having or being predisposed to a disease, disorder, or condition associated with PNPLA3 expression, such as a biomarker (e.g., PNPLA3 mRNA, PNPLA3 protein, or a combination thereof). Similarly, as detailed below, the method also includes additional steps such as measuring or obtaining baseline values ​​of a PNPLA3 expression marker (e.g., PNPLA3 protein) and then evaluating the effectiveness of the treatment by comparing the thus obtained value with one or more other baseline values ​​or values ​​obtained after administering one or more oligonucleotides to the individual. [Examples]

[0200] The following non-limiting embodiments are presented for illustrative purposes only and are not limiting.

[0201] Synthesis of oligonucleotides

[0202] Example 1: Preparation of dsRNAi oligonucleotides

[0203] Synthesis and Purification of Oligonucleotides: The dsRNAi oligonucleotides in the examples are chemically synthesized using the methods described herein. Generally, dsRNAi oligonucleotides are synthesized using solid-phase oligonucleotide synthesis methods described for 19-23mer siRNAs (see, for example, Scaringe et al. (1990) Nucleic Acids Res. 18:5433-41 and Usman et al. (1987) J.Am. Chem. Soc. 109:7845-45; also see U.S. Patents 5,804,683; 5,831,071; 5,998,203; 6,008,400; 6,111,086; 6,117,657; 6,353,098; 6,362,323; 6,437,117; and 6,469,158).

[0204] Individual RNA strands are synthesized and purified by HPLC according to the standard method (Integrated DNA Technologies). For example, RNA oligonucleotides are synthesized using a solid-phase phosphoramidite reaction, deprotected, and desalted on an NAP-5 column (Amersham Pharmacia Biotech; Piscataway, NJ) using the standard technique (Damha & Olgivie (1993) Methods Mol. Biol. 20:81-114; Wincott et al. (1995) Nucleic Acids Res. 23:2677-84). The oligomers are purified by ion-exchange high-performance liquid chromatography (IE-HPLC) on an Amersham Source 15Q column (1.0 cm × 25 cm; Amersham Pharmacia Biotech) using a stepwise linear gradient for 15 minutes. The gradient changes from 90:10 buffer A:B to 52:48 buffer A:B, where buffer A is 100 mM Tris at pH 8.5, and buffer B is 100 mM Tris and 1 M NaCl at pH 8.5. The sample is monitored at 260 nm, peaks corresponding to full-length oligonucleotide species are collected and pooled, desalted on an NAP-5 column, and lyophilized.

[0205] The purity of each oligomer is determined by capillary electrophoresis (CE) on a Beckman PACE 5000 (Beckman Coulter, Inc.). The CE capillary has an inner diameter of 100 μm and contains ssDNA 100R Gel (Beckman-Coulter). Typically, an oligonucleotide of approximately 0.6 nmole is injected into the capillary, electrophoresed in an electric field of 444 V / cm, and detected by UV absorbance at 260 nm. Denatured Tris-borate-7M-urea electrophoresis buffer is purchased from Beckman-Coulter. Oligo-ribonucleotides of at least 90% purity, as evaluated by CE, are obtained for use in the experiments described below. The identity of the compounds is verified by matrix-assisted laser desorption / ionization time-of-flight (MALDI-TOF) mass spectrometry on a Voyager DE® Biospectometry Work Station (Applied Biosystems) according to the manufacturer's recommended protocol. In most cases, the relative molecular weight of all oligomers can be obtained within 0.2% of the expected molecular weight.

[0206] Preparation of double helical molecules: Resuspend the ssRNA oligomer in a double helical buffer containing 100 mM potassium acetate, 30 mM HEPES, and pH 7.5 (e.g., at a concentration of 100 μM). Mix the complementary sense and antisense strands in equimolar amounts to obtain a final double helical solution, e.g., 50 μM. Heat the sample in RNA buffer (IDT) to 100°C for 5 minutes and cool to room temperature before use. dsRNA oligonucleotides are stored at -20°C. ssRNA oligomers are stored lyophilized or in nuclease-free water at -80°C.

[0207] Functional in vitro

[0208] Example 2: RNAi oligonucleotide inhibition of PNPLA3 expression in vitro

[0209] Identification of PNPLA3 Target Sequences: To identify RNAi oligonucleotide inhibitors of PNPLA3 expression, a computer-based algorithm was used to computer-generate PNPLA3 target sequences suitable for assaying inhibition of PNPLA3 expression via the RNAi pathway. The algorithm provides RNAi oligonucleotide guide (antisense) strand sequences complementary to suitable PNPLA3 target sequences in human PNPLA3 mRNA (e.g., SEQ ID NO: 1). Some of the guide strand sequences identified by the algorithm are also complementary to the corresponding PNPLA3 target sequences in monkey PNPLA mRNA. Some of the antisense (guide) strand sequences identified by the algorithm are also complementary to the corresponding PNPLA3 target sequences in monkey PNPLA3 mRNA (e.g., SEQ ID NO: 5). From this, 384 dsRNAi oligonucleotides (formatted as DsiRNA oligonucleotides) were generated, each possessing a unique antisense strand with a complementary region to the PNPLA3 target sequences identified by the algorithm.

[0210] [Table 1-1] [Table 1-2] [Table 1-3] [Table 1-4] [Table 1-5] [Table 1-6] [Table 1-7] [Table 1-8] [Table 1-9] [Table 1-10] [Table 1-11] [Table 1-12] [Table 1-13] [Table 1-14]

[0211] In vitro cell-based assays: The ability of each of the 384 DsiRNAs listed in Table 1 to inhibit PNPLA3 expression is determined using in vitro cell-based assays. Furthermore, the nucleotide sequences of the passenger and guide strands of the DsiRNAs have a unique pattern of modified nucleotides and phosphorothioate bonds (see, for example, Figure 2). Briefly, HuH-7 human liver cells stably expressing PNPLA3 are transfected with each DsiRNA (0.5 nM) in separate wells of a multi-well cell culture plate. The cells are maintained for 24 hours post-transfection, and then the level of residual PNPLA3 mRNA from the transfected cells is determined using a TAQMAN®-based qPCR assay. Two qPCR assays, a 3' assay and a 5' assay, are used to determine mRNA levels measured by HEX probes and FAM probes, respectively.

[0212] Table 2 shows the results of HuH-7 cell-based assays using DsiRNA. Table 2 shows the results of HuH-7 cell-based assays using 384 DsiRNAs with guide strands complementary to human and monkey PNPLA3 mRNA ("double common"). Transfection with double common DsiRNAs that resulted in less than 30% of the PNPLA3 mRNA remaining in cells compared to a negative control is considered a candidate PNPLA3 expression inhibitor (referred to herein as a "hit"). [Table 2-1] [Table 2-2] [Table 2-3] [Table 2-4] [Table 2-5] [Table 2-6] [Table 2-7] [Table 2-8] [Table 2-9]

[0213] These results demonstrate that DsiRNAs designed to target human PNPLA3 mRNA inhibit PNPLA3 expression in cells (determined by a decrease in PNPLA3 mRNA levels in DsiRNA-transfected cells), and that nucleotide sequences containing DsiRNA hits are useful for generating RNAi oligonucleotides that inhibit PNPLA3 expression. Furthermore, these results indicate that multiple PNPLA3 target sequences are suitable for RNAi-mediated inhibition of PNPLA3 expression.

[0214] Function in vivo Example 3: RNAi oligonucleotide inhibition of PNPLA3 expression in vivo Of the 384 DsiRNAs screened using the HuH-7 cell-based assay described in Example 2, nucleotide sequences of 192 DsiRNA hits (Table 3) were selected for further evaluation in vivo. Briefly, the nucleotide sequences of the selected DsiRNAs were used to generate 194 corresponding dsRNAi oligonucleotides containing nicked tetraloop GalNAc conjugate structures (referred herein to as "GalNAc-conjugate PNPLA3 oligonucleotides") having a 36-mer sense (passenger) strand and a 22-mer antisense (guide) strand. Furthermore, the nucleotide sequences of the passenger and guide strands of the GalNAc-conjugate PNPLA3 oligonucleotides have a unique pattern of modified nucleotides and phosphorothioate bonds (see, for example, Figure 1 for a schematic diagram of the general structure and chemical modification pattern of the GalNAc-conjugate PNPLA3 oligonucleotide). The three adenosine nucleotides of the tetraloop are each conjugated to the GalNAc moiety (CAS number: 14131-60-3). [Table 3-1] [Table 3-2] [Table 3-3] [Table 3-4] [Table 3-5]

[0215] Mouse Tests: Various GalNAc-conjugated PNPLA3 oligonucleotides (some of which are listed in Table 3) were evaluated in a hydrodynamic injection (HDI) mouse model. Additional HDI tests are listed in Tables 4, 5, and 6. In these HDI tests, mice were engineered to transiently express human PNPLA3 mRNA in hepatocytes. GalNAc-conjugated PNPLA3 oligonucleotide controls (SEQ ID NOs. 1165 and 1166) were used as benchmark controls. Briefly, 6-8 week old female CD-1 mice were subcutaneously treated with GalNAc-conjugated PNPLA3 oligonucleotides at a dose level of 1 mg / kg. Three days (72 hours) later, a DNA plasmid encoding the complete human PNPLA3 gene was hydrodynamically injected into the mice under the control of a ubiquitous cytomegalovirus (CMV) promoter sequence. Liver samples were collected one day after plasmid introduction. Total RNA from these mice is subjected to qRT-PCR analysis for PNPLA3 mRNA and compared to mice treated with the same amount of PBS alone. The values ​​are normalized for transfection efficiency using the NeoR gene contained in the plasmid.

[0216] As shown in Tables 4, 5, and 6, the number of tested GalNAc-conjugate PNPLA3 oligonucleotides that inhibited PNPLA3 expression, determined by the reduction in PNPLA3 mRNA levels in liver samples from oligonucleotide-treated mice, was compared to that of mice treated with PBS. The mean residual PNPLA3 mRNA percentage (%) in liver samples from mice treated with a benchmark GalNAc-conjugate PNPLA3 oligonucleotide control was compared to that of mice treated with PBS. Table 4 shows that 18 out of 21 GalNAc-conjugate PNPLA3 oligonucleotides significantly inhibited PNPLA3 expression more than the reference GalNAc-conjugate PNPLA3 oligonucleotide used as a control. The sequences of these oligonucleotides, along with their modification patterns and sequence numbers, are disclosed in Table A. [Table 4]

[0217] Tables 5 and 6 show two additional HDI test sets, each using 18 GalNAc-conjugated PNPLA3 oligonucleotides and the same reference oligonucleotide. The sequences of these oligonucleotides, along with their modification patterns and sequence numbers, are disclosed in Tables B and C, respectively.

[0218] Based on these results, ten GalNAc-conjugated PNPLA3 oligonucleotides were selected to evaluate their ability to inhibit PNPLA3 expression in non-human primates (NHPs). The GalNAc-conjugated PNPLA3 oligonucleotides have chemically modified nucleotides in the pattern shown in Figure 1. [Table 5] [Table 6]

[0219] Table 7 discloses additional HDI testing using the best GalNAc-conjugated PNPLA3 oligonucleotide (hit) based on the results obtained from the prior tests and three prior HDI screenings disclosed in Tables 4, 5, and 6. The sequences of the oligonucleotides used in this HDI screening, along with their modification patterns and sequence numbers, are disclosed in Table D. [Table 7]

[0220] NHP Study: Ten GalNAc-conjugated PNPLA3 oligonucleotides selected from Table 7 were evaluated in cynomolgus monkeys (Macaca fascicularis). In this study, NHPs were grouped so that the mean body weight (approximately 5.4 kg) was comparable between the control and experimental groups. Each cohort included two male subjects and three female subjects. GalNAc-conjugated PNPLA3 oligonucleotides were administered subcutaneously on day 0 of the study. Blood samples were collected on days 8, 5, and 0 of the study, and weekly thereafter. Ultrasound-guided core-needle liver biopsies were collected on days 7, 28, and 56 of the study. At each time point, total RNA from liver biopsy samples was subjected to qRT-PCR analysis, and PNPLA3 mRNA in oligonucleotide-treated monkeys was measured and compared with monkeys treated with an equivalent amount of PBS. To normalize the data, measurements were performed against the geometric mean of two reference genes, PPIB and 18S rRNA. As shown in Tables 8 (-7 days), 9 (28 days), and 10 (56 days), treatment of NHP with GalNAc-conjugated PNPLA3 oligonucleotides inhibits PNPLA3 expression in the liver, as determined by a decrease in PNPLA3 mRNA levels in liver samples derived from NHP treated with oligonucleotides, compared to NHP treated with PBS. At all time points evaluated, PNPLA3 oligonucleotides 34, 81, and 121 significantly inhibited PNPLA3 expression compared to benchmark PBS time-matched controls. From the same NHP studies, inhibition of PNPLA3 expression was also determined by measuring PNPLA3 protein in serum prepared from pre-administration and weekly blood samples by ELISA. In summary, these results demonstrate that treatment of NHP with GalNAc-conjugated PNPLA3 oligonucleotides reduces PNPLA3 mRNA levels in the liver and simultaneously reduces PNPLA3 protein levels in serum.

[0221] [Table 8]

[0222]

Table 9

Table 10

[0223] In summary, these results indicate that GalNAc-conjugated PNPLA3 oligonucleotides designed to target human PNPLA3 mRNA inhibit PNPLA3 expression in vivo (determined by the decrease in the amount of PNPLA3 mRNA and PNPLA3 protein in treated animals). Another aspect of the present invention may be as follows: [1] An RNAi oligonucleotide for reducing the expression of PNPLA3, wherein the oligonucleotide comprises a sense strand and an antisense strand, the sense strand and the antisense strand form a double-stranded region, the antisense strand comprises a complementary region to one of the PNPLA3 mRNA target sequences of sequence numbers 1167 to 1176, and the complementary region has a length of at least 15 consecutive nucleotides. [2] The RNAi oligonucleotide described in [1], wherein the sense strand is 15 to 50 nucleotides long. [3] The RNAi oligonucleotide according to [1] or [2], wherein the sense strand is 18 to 36 nucleotides long. [4] The RNAi oligonucleotide described in any one of the above [1] to [3], wherein the antisense strand is 15 to 30 nucleotides long. [5] The RNAi oligonucleotide according to any one of [1] to [4], wherein the antisense strand is 22 nucleotides long, and the antisense strand and the sense strand form a double-stranded region of at least 19 nucleotides long, and optionally at least 20 nucleotides long. [6] The RNAi oligonucleotide according to any one of [1] to [5] above, wherein the complementary region has a length of at least 19 consecutive nucleotides, and optionally at least 20 nucleotides. [7] The RNAi oligonucleotide according to any one of [1] to [6], wherein the 3' end of the sense strand includes a stem loop represented as S1-L-S2, where S1 is complementary to S2, and L forms a loop of 3 to 5 nucleotides between S1 and S2. [8] RNAi oligonucleotide for reducing PNPLA3 expression, wherein the oligonucleotide comprises a sense strand of 15 to 50 nucleotides in length and an antisense strand, wherein the sense strand and the antisense strand form a double-stranded region, the antisense strand includes a complementary region to any one of the PNPLA3 mRNA target sequences of sequence numbers 1167 to 1176, and the complementary region is at least 15 consecutive nucleotides in length. [9] An RNAi oligonucleotide for reducing the expression of PNPLA3, wherein the oligonucleotide comprises a sense strand of 15 to 50 nucleotides in length and an antisense strand of 15 to 30 nucleotides in length, the sense strand and the antisense strand form a double-stranded region, the antisense strand comprises a complementary region to any one of the PNPLA3 mRNA target sequences of sequence numbers 1167 to 1176, and the complementary region is at least 15 consecutive nucleotides in length.

[10] RNAi oligonucleotide for reducing PNPLA3 expression, wherein the oligonucleotide comprises a sense strand and an antisense strand of 15 to 50 nucleotides in length, the sense strand and the antisense strand form a double-stranded region, the antisense strand includes a complementary region for any one of the PNPLA3 mRNA target sequences of sequence numbers 1167 to 1176, and the complementary region is 19 consecutive nucleotides long, or optionally 20 nucleotides long.

[11] RNAi oligonucleotide for reducing PNPLA3 expression, wherein the oligonucleotide comprises a sense strand and an antisense strand of 18 to 36 nucleotides in length, the sense strand and the antisense strand form a double-stranded region, the antisense strand comprises a complementary region to one of the PNPLA3 mRNA target sequences of sequence numbers 1167 to 1176, and the complementary region is 19 consecutive nucleotides long, or optionally 20 nucleotides long.

[12] RNAi oligonucleotide for reducing PNPLA3 expression, wherein the oligonucleotide comprises a sense strand of 18 to 36 nucleotides and an antisense strand of 22 nucleotides, the sense strand and the antisense strand form a double-stranded region, the antisense strand comprises a complementary region to one of the PNPLA3 mRNA target sequences of sequence numbers 1167 to 1176, and the complementary region is 19 consecutive nucleotides long, or optionally 20 nucleotides long.

[13] RNAi oligonucleotide for reducing PNPLA3 expression, wherein the oligonucleotide comprises a sense strand of 18 to 36 nucleotides in length and an antisense strand of 22 nucleotides in length, the sense strand and the antisense strand form a double-stranded region, the 3' end of the sense strand comprises a stem-loop represented as S1-L-S2 (S1 is complementary to S2, and L forms a loop of 3 to 5 nucleotides in length between S1 and S2), the antisense strand comprises a complementary region to one of the PNPLA3 mRNA target sequences of sequence numbers 1167 to 1176, and the complementary region is 19 consecutive nucleotides in length, or optionally 20 nucleotides in length.

[14] RNAi oligonucleotide for reducing PNPLA3 expression, wherein the oligonucleotide comprises a sense strand of 36 nucleotides and an antisense strand of 22 nucleotides, the sense strand and the antisense strand form a double-stranded region, the 3' end of the sense strand comprises a stem-loop represented as S1-L-S2 (S1 is complementary to S2, and L forms a loop of 3-5 nucleotides between S1 and S2), the antisense strand comprises a complementary region to one of the PNPLA3 mRNA target sequences of sequence numbers 1167-1176, and the complementary region is 19 consecutive nucleotides long, or optionally 20 nucleotides long.

[15] RNAi oligonucleotide for reducing PNPLA3 expression, wherein the oligonucleotide comprises a sense strand of 36 nucleotides and an antisense strand of 22 nucleotides, wherein the sense strand and the antisense strand form a double-stranded region of at least 19 nucleotides, optionally 20 nucleotides, and the 3' end of the sense strand comprises a stem-loop represented as S1-L-S2 (S1 is complementary to S2, and L forms a loop of 3-5 nucleotides between S1 and S2), and the antisense strand comprises a complementary region to one of the PNPLA3 mRNA target sequences of sequence numbers 1167-1176, wherein the complementary region is 19 consecutive nucleotides, optionally 20 nucleotides long.

[16] RNAi oligonucleotides according to any one of the above items [7] and

[13] to

[15] , wherein L is a triloop or a tetraloop.

[17] The RNAi oligonucleotide described in

[16] above, wherein L is a tetraloop.

[18] The RNAi oligonucleotide according to

[17] , wherein the tetraloop comprises the sequence 5'-GAAA-3'.

[19] RNAi oligonucleotides according to any one of the above

[16] to

[18] , wherein S1 and S2 are 1 to 10 nucleotides in length and have the same length.

[20] The RNAi oligonucleotide according to

[19] , wherein S1 and S2 are 1 nucleotide, 2 nucleotides, 3 nucleotides, 4 nucleotides, 5 nucleotides, 6 nucleotides, 7 nucleotides, 8 nucleotides, 9 nucleotides, or 10 nucleotides in length.

[21] The RNAi oligonucleotide described in

[20] , wherein S1 and S2 are 6 nucleotides long.

[22] The RNAi oligonucleotide according to any one of

[16] to

[21] , wherein the stem-loop comprises the sequence 5'-GCAGCCGAAAGGCUGC-3' (sequence number 1177).

[23] The RNAi oligonucleotide according to any one of [1] to

[22] , wherein the antisense strand includes a 3' overhang sequence of 1 nucleotide length or longer.

[24] The RNAi oligonucleotide according to

[23] , wherein the 3' overhang sequence is 2 nucleotides long, and optionally the 3' overhang sequence is GG.

[25] The RNAi oligonucleotide according to any one of the prior embodiments, wherein the oligonucleotide comprises at least one modified nucleotide.

[26] The RNAi oligonucleotide according to

[25] , wherein the modified nucleotide includes a 2'-modification.

[27] The RNAi oligonucleotide according to

[26] , wherein the 2'-modification is a modification selected from the group consisting of 2'-aminoethyl, 2'-fluoro, 2'-O-methyl, 2'-O-methoxyethyl, and 2'-deoxy-2'-fluoro-β-d-arabinonucleotide.

[28] The RNAi oligonucleotide according to any one of the above

[25] to

[27] , wherein all nucleotides constituting the oligonucleotide are modified, and the modification is optionally a 2'-modification selected from the group consisting of 2'-fluoro and 2'-O-methyl.

[29] The RNAi oligonucleotide according to any one of the prior embodiments, wherein the oligonucleotide comprises at least one modified nucleotide bond.

[30] The RNAi oligonucleotide according to

[29] , wherein the at least one modified nucleotide bond is a phosphorothioate bond.

[31] The RNAi oligonucleotide according to any one of the prior embodiments, wherein the 4'-carbon of the sugar of the 5'-nucleotide of the antisense chain contains a phosphate analog.

[32] The RNAi oligonucleotide according to

[31] , wherein the phosphate analog is an oxymethylphosphonate, a vinylphosphonate, or a malonylphosphonate, and optionally the phosphate analog is a 4'-phosphate analog containing 5'-methoxyphosphonate-4'-oxy.

[33] The RNAi oligonucleotide according to any one of the prior embodiments, wherein at least one nucleotide of the oligonucleotide is conjugated to one or more targeting ligands.

[34] The RNAi oligonucleotide described in

[33] , wherein each targeting ligand comprises a carbohydrate, amino sugar, cholesterol, polypeptide, or lipid.

[35] The RNAi oligonucleotide according to

[33] , wherein each targeted ligand contains an N-acetylgalactosamine (GalNAc) moiety.

[36] The RNAi oligonucleotide according to

[35] , wherein the GalNAc portion is a monovalent GalNAc portion, a divalent GalNAc portion, a trivalent GalNAc portion, or a tetravalent GalNAc portion.

[37] The RNAi oligonucleotide according to any one of the above

[16] to

[32] , wherein up to four nucleotides of the stem-loop L are each conjugated to a monovalent GalNAc portion.

[38] The sense chain is sequence numbers 9, 11, 13, 15, 17, 19, 21, 23, 25, 27, 29, 31, 33, 35, 37, 39, 41, 43, 45, 47, 49, 51, 53, 55, 57, 59, 61, 63, 65, 67, 69, 71, 73, 75, 77, 79, 81, 83, 85, 87, 89, 91, 93, 95, 97, 99, 101, 103, 105, 107, 109, 111, 113, 115, 117, 119, 121, 123, 125, 127, 129, 131, 133, 135, 137, 139, 141, 143, 145, 147, 1 49, 151, 153, 155, 157, 159, 161, 163, 165, 167, 169, 171, 173, 175, 177, 179, 181, 183, 185, 187, 189, 191, 193, 195, 197, 199, 201, 203, 205, 207, 209, 21 1, 213, 215, 217, 219, 221, 223, 225, 227, 229, 231, 233, 235, 237, 239, 241, 243, 245, 247, 249, 251, 253, 255, 257, 259, 261, 263, 265, 267, 269, 271, 273 ,275,277,279,281,283,285,287,289,291,293,295,297,299,301,303,305,307,309,311,313,315,317,319,321,323,325,327,329,331,333,335, 337, 339, 341, 343, 345, 347, 349, 351, 353, 355, 357, 359, 361, 363, 365, 367, 369, 371, 373, 375, 377, 379, 381, 383, 385, 387, 389, 391, 393, 395, 397, 3 99, 401, 403, 405, 407, 409, 411, 413, 415, 417, 419, 421, 423, 425, 427, 429, 431, 433, 435, 437, 439, 441, 443, 445, 447, 449, 451, 453, 455, 457, 459, 46 1, 463, 465, 467, 469, 471, 473, 475, 477, 479, 481, 483, 485, 487, 489, 491, 493, 495, 497, 499, 501, 503, 505, 507, 509, 511, 513, 515, 517, 519, 521, 523,525, 527, 529, 531, 533, 535, 537, 539, 541, 543, 545, 547, 549, 551, 553, 555, 557, 559, 561, 563, 565, 567, 569, 571, 573, 575, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, 629, 631, 633, 635, 637, 639, 641, 643, 645, 647, 649, 651, 653, 655, 657, 659, 661, 663, 665, 667, 669, 671, 673, 675, 677, 679, 681, 683, 685, 687, 689, 691, 693, 695, 697, 699, 701, 703, 705, 707, 709, 711, 713, 715, 717, 719, 721, Contains any one of the nucleotide sequences 723, 725, 727, 729, 731, 733, 735, 737, 739, 741, 743, 745, 747, 749, 751, 753, 755, 757, 759, 761, 763, 765, 767, 769, 771, 773, and 775, or The aforementioned sense chains are sequence numbers 777, 779, 781, 783, 785, 787, 789, 791, 793, 795, 797, 799, 801, 803, 805, 807, 809, 811, 813, 815, 817, 819, 821, 823, 825, 827, 829, 831, 833, 835, 837, 839, 841, 843, 845, 847, 849, 851, 853, 855, 857, 859, 861, 863, 865, 867, 869, 871, 873, 875, 877, 879, 881, 883, 885, 887, 889, 891, 893, 89 5, 897, 899, 901, 903, 905, 907, 909, 911, 913, 915, 917, 919, 921, 923, 925, 927, 929, 931, 933, 935, 937, 939, 941, 943, 945, 947, 949, 951, 953, 955, 957 ,959,961,963,965,967,969,971,973,975,977,979,981,983,985,987,989,991,993,995,997,999,1001,1003,1005,1007,1009,1011,1013,1015, 1017, 1019, 1021, 1023, 1025, 1027, 1029, 1031, 1033, 1035, 1037, 1039, 1041, 1043, 1045, 1047, 1049, 1051, 1053, 1055, 1057, 1059, 1061, 1063, 1065, 1067, 1069, 1071, 1073, 1075, 1077, 1079, 1081, 1083, 1085, 1087, 1089, 1091, 1093, 1095, 1097, 1099, 1101, 1103, 1105, 1107, 1109, 1111, 1113, 1115, 1117, 1119, 1121, 1123, 1125, 1127, 1129, 1131, 1133, 1135, 1137, 1139, 1141, 1143, 1145, 1147, 1149, 1151, 1153, 1155, 1157, 1159, 1161, and 1163, or the sense strand contains any one nucleotide sequence, or sequence numbers 1178, 1180, 1182, 1184, 1186, 1188, 1190, 1192, 1194, 1196, 1198, 1200, 1202, 1204, 1206, 1208, 1210, 1212, 1214,Contains one of the following nucleotide sequences: 1216, 1218, 1220, 1222, 1224, 1226, 1228, 1230, 1232, 1234, 1236, 1238, 1240, 1242, 1244, 1246, 1248, 1250, 1252, 1254, 1256, 1258, 1260, 1262, 1264, 1266, 1268, 1270, 1272, 1274, 1276, 1278, 1280, 1282, 1284, 1286, 1288, 1290, 1292, 1294, 1296, 1298, or 1300. The RNAi oligonucleotide described in any one of the above items [1] to

[37] .

[39] The antisense chain is sequence numbers 10, 12, 14, 16, 18, 20, 22, 24, 26, 28, 30, 32, 34, 36, 38, 40, 42, 44, 46, 48, 50, 52, 54, 56, 58, 60, 62, 64, 66, 68, 70, 72, 74, 76, 78, 80, 82, 84, 86, 88, 90, 92, 94, 96, 98, 100, 102, 104, 106, 108, 110, 112, 114, 116, 118, 120, 122, 124, 126, 128, 130, 132, 134, 136, 138, 140, 142, 144, 146 ,148,150,152,154,156,158,160,162,164,166,168,170,172,174,176,178,180,182,184,186,188,190,192,194,196,198,200,202,204,206,208, 210, 212, 214, 216, 218, 220, 222, 224, 226, 228, 230, 232, 234, 236, 238, 240, 242, 244, 246, 248, 250, 252, 254, 256, 258, 260, 262, 264, 266, 268, 270, 2 72, 274, 276, 278, 280, 282, 284, 286, 288, 290, 292, 294, 296, 298, 300, 302, 304, 306, 308, 310, 312, 314, 316, 318, 320, 322, 324, 326, 328, 330, 332, 3 34, 336, 338, 340, 342, 344, 346, 348, 350, 352, 354, 356, 358, 360, 362, 364, 366, 368, 370, 372, 374, 376, 378, 380, 382, ​​384, 386, 388, 390, 392, 394, 39 6, 398, 400, 402, 404, 406, 408, 410, 412, 414, 416, 418, 420, 422, 424, 426, 428, 430, 432, 434, 436, 438, 440, 442, 444, 446, 448, 450, 452, 454, 456, 458 ,460,462,464,466,468,470,472,474,476,478,480,482,484,486,488,490,492,494,496,498,500,502,504,506,508,510,512,514,516,518,520,522, 524, 526, 528, 530, 532, 534, 536, 538, 540, 542, 544, 546, 548, 550, 552, 554, 556, 558, 560, 562, 564, 566, 568, 570, 572, 574, 576, 578, 580, 582, 584, 586, 58 8, 590, 592, 594, 596, 598, 600, 602, 604, 606, 608, 610, 612, 614, 616, 618, 620, 622, 624, 626, 628, 630, 632, 634, 636, 638, 640, 642, 644, 646, 648, 650, 652, 654, 656, 658, 660, 662, 664, 666, 668, 670, 672, 674, 676, 678, 680, 682, 684, 686, 688, 690, 692, 694, 696, 698, 700, 702, 704, 706, 708, 710, 712, 714, 716, 718, 720, 72 Contains any one of the nucleotide sequences 2, 724, 726, 728, 730, 732, 734, 736, 738, 740, 742, 744, 746, 748, 750, 752, 754, 756, 758, 760, 762, 764, 766, 768, 770, 772, 774, and 776, or The aforementioned antisense chains are sequence numbers 778, 780, 782, 784, 786, 788, 790, 792, 794, 796, 798, 800, 802, 804, 806, 808, 810, 812, 814, 816, 818, 820, 822, 824, 826, 828, 830, 832, 834, 836, 838, 840, 842, 844, 846, 848, 850, 852, 854, 856, 858, 860, 862, 864, 866, 868, 870, 872, 874, 876, 878, 880, 882, 884, 886, 888, 890, 892, 894, 896, 898, 900, 902, 904, 906, 908, 910, 912, 914, 916, 918, 920, 922, 924, 926, 928, 930, 932, 934, 936, 938, 940, 942, 944, 946, 948, 950, 952, 954, 956, 958, 960, 962, 964, 966, 968, 970, 972, 974, 976, 978, 980, 982, 984, 986, 988, 990, 992, 994, 996, 998, 1000, 1002, 1004, 1006, 1008, 1010, 1012, 1014, 1016, 1018, 1020, 1022, 1024, 1026, 1028, 1030, 1032, 1034, 1036, 1038, 1040, 1042, 1044, 1046, 1048, 1050, 1052, 1054, 1056, 1058, 1060, 1062, 1064, 1066, 1068, 1070, 1072, 1074, 1076, 1078, 1080, 1082, 10 Contains one of the following nucleotide sequences: 84, 1086, 1088, 1090, 1092, 1094, 1096, 1098, 1100, 1102, 1104, 1106, 1108, 1110, 1112, 1114, 1116, 1118, 1120, 1122, 1124, 1126, 1128, 1130, 1132, 1134, 1136, 1138, 1140, 1142, 1144, 1146, 1148, 1150, 1152, 1154, 1156, 1158, 1160, 1162, and 1164; Or the antisense chain is sequence numbers 1179, 1181, 1183, 1185, 1187, 1189, 1191, 1193, 1195, 1197, 1199, 1201, 1203, 1205, 1207, 1209, 1211, 1213, 1215, 1217, 1219, 1221, 1223, 1225, 1227, 1229, 1231, 1233, 1235, 1237, 1239, 12 Contains one of the following nucleotide sequences: 41, 1243, 1245, 1247, 1249, 1251, 1253, 1255, 1257, 1259, 1261, 1263, 1265, 1267, 1269, 1271, 1273, 1275, 1277, 1279, 1281, 1283, 1285, 1287, 1289, 1291, 1293, 1295, 1297, 1299, or 1301. The RNAi oligonucleotide described in any one of the above items [1] to

[38] .

[40] The sense chain and the antisense chain are as follows: (a) Sequence IDs 787 and 788, (b) Sequence IDs 843 and 844, (c) Sequence IDs 867 and 868, (d) Sequence IDs 871 and 872, (e) Sequence IDs 937 and 938, (f) Sequence IDs 1003 and 1004, (g) Sequence IDs 1007 and 1008, (h) Sequence IDs 1017 and 1018, (i) Sequence IDs 1161 and 1162, (j) Sequence IDs 1163 and 1164, (k) Sequence IDs 1220 and 1221, (l) Sequence IDs 1224 and 1225, (m) Sequence IDs 1230 and 1231, (n) Sequence IDs 1232 and 1233, (o) Sequence IDs 1188 and 1189, (p) Sequence IDs 1190 and 1191, (q) Sequence IDs 1244 and 1245, (r) Sequence IDs 1250 and 1251-, (s) Sequence IDs 1254 and 1255, and (t) Sequence IDs 1246 and 1247 An RNAi oligonucleotide according to any one of the above [1] to

[39] , comprising a nucleotide sequence selected from the group consisting of the above.

[41] The RNAi oligonucleotide according to any one of [1] to

[39] , wherein the sense strand comprises the nucleotide sequence described in SEQ ID NO: 1220 and the antisense strand comprises the nucleotide sequence described in SEQ ID NO: 1221.

[42] The RNAi oligonucleotide according to any one of [1] to

[39] , wherein the sense strand comprises the nucleotide sequence described in SEQ ID NO: 1224 and the antisense strand comprises the nucleotide sequence described in SEQ ID NO: 1225.

[43] The RNAi oligonucleotide according to any one of [1] to

[39] , wherein the sense strand comprises the nucleotide sequence described in SEQ ID NO: 1230 and the antisense strand comprises the nucleotide sequence described in SEQ ID NO: 1231.

[44] The RNAi oligonucleotide according to any one of the above [1] to

[39] , wherein the sense strand comprises the nucleotide sequence described in SEQ ID NO: 1232 and the antisense strand comprises the nucleotide sequence described in SEQ ID NO: 1233.

[45] The RNAi oligonucleotide according to any one of [1] to

[39] , wherein the sense strand comprises the nucleotide sequence described in SEQ ID NO: 1188 and the antisense strand comprises the nucleotide sequence described in SEQ ID NO: 1189.

[46] The RNAi oligonucleotide according to any one of [1] to

[39] , wherein the sense strand comprises the nucleotide sequence described in SEQ ID NO: 1190 and the antisense strand comprises the nucleotide sequence described in SEQ ID NO: 1191.

[47] The RNAi oligonucleotide according to any one of [1] to

[39] , wherein the sense strand comprises the nucleotide sequence described in SEQ ID NO: 1244 and the antisense strand comprises the nucleotide sequence described in SEQ ID NO: 1245.

[48] ​​The RNAi oligonucleotide according to any one of the above [1] to

[39] , wherein the sense strand comprises the nucleotide sequence described in SEQ ID NO: 1250 and the antisense strand comprises the nucleotide sequence described in SEQ ID NO: 1251.

[49] The RNAi oligonucleotide according to any one of [1] to

[39] , wherein the sense strand comprises the nucleotide sequence described in SEQ ID NO: 1254 and the antisense strand comprises the nucleotide sequence described in SEQ ID NO: 1255.

[50] The RNAi oligonucleotide according to any one of [1] to

[39] , wherein the sense strand comprises the nucleotide sequence described in SEQ ID NO: 1246 and the antisense strand comprises the nucleotide sequence described in SEQ ID NO: 1247.

[51] RNAi oligonucleotide for reducing PNPLA3 expression, wherein the oligonucleotide comprises a sense strand and an antisense strand, the sense strand and the antisense strand form a double-stranded region, all nucleotides constituting the sense strand and the antisense strand are modified, the antisense strand includes a complementary region to any one of the PNPLA3 mRNA target sequences of sequence numbers 1167 to 1176, and the complementary region has a length of at least 15 consecutive nucleotides.

[52] RNAi oligonucleotide for reducing PNPLA3 expression, wherein the oligonucleotide comprises a sense strand and an antisense strand, the sense strand and the antisense strand form a double-stranded region, all nucleotides constituting the sense strand and the antisense strand are modified, the 4'-carbon of the sugar of the 5'-nucleotide of the antisense strand comprises a phosphate analog, the antisense strand comprises a complementary region to any one of the PNPLA3 mRNA target sequences of SEQ ID NOs. 1167 to 1176, and the complementary region has a length of at least 15 consecutive nucleotides.

[53] RNAi oligonucleotide for reducing PNPLA3 expression, wherein the oligonucleotide comprises a sense strand and an antisense strand, the sense strand and the antisense strand form a double-stranded region, all nucleotides constituting the sense strand and the antisense strand are modified, the 4'-carbon of the sugar of the 5'-nucleotide of the antisense strand comprises a phosphate analog, the antisense strand comprises a complementary region to any one of the PNPLA3 mRNA target sequences of SEQ ID NOs. 1167 to 1176, and the complementary region has a length of at least 15 consecutive nucleotides.

[54] RNAi oligonucleotide for reducing PNPLA3 expression, wherein the oligonucleotide comprises a sense strand and an antisense strand, the sense strand and the antisense strand form a double-stranded region, all nucleotides constituting the sense strand and the antisense strand are modified, the antisense strand and the sense strand comprise one or more 2'-fluoro and 2'-O-methyl modified nucleotides and at least one phosphorothioate bond, the 4'-carbon of the sugar of the 5'-nucleotide of the antisense strand comprises a phosphate analog, the antisense strand comprises a complementary region to any one of the PNPLA3 mRNA target sequences of SEQ ID NOs. 1167 to 1176, and the complementary region has a length of at least 15 consecutive nucleotides.

[55] The RNAi oligonucleotide according to any one of the above

[51] to

[54] , wherein the sense strand is composed of any one of sequence numbers 787, 843, 867, 871, 937, 1003, 1007, 1017, 1161, or 1163, or the sense strand is composed of any one of sequence numbers 1188, 1190, 1220, 1224, 1230, 1232, 1244, 1246, 1250, or 1254.

[56] The RNAi oligonucleotide according to any one of the above

[51] to

[55] , wherein the antisense strand is composed of any one of sequence numbers 788, 844, 868, 872, 938, 1004, 1008, 1018, 1162, or 1164, or the antisense strand is composed of any one of sequence numbers 1189, 1191, 1221, 1225, 1231, 1233, 1245, 1247, 1251, or 1255.

[57] The sense chain and the antisense chain are as follows: (a) Sequence IDs 787 and 788, (b) Sequence IDs 843 and 844, (c) Sequence IDs 867 and 868, (d) Sequence IDs 871 and 872, (e) Sequence IDs 937 and 938, (f) Sequence IDs 1003 and 1004, (g) Sequence IDs 1007 and 1008, (h) Sequence IDs 1017 and 1018, (i) Sequence IDs 1161 and 1162, and (j) Sequence IDs 1163 and 1164, (k) Sequence IDs 1220 and 1221, (l) Sequence IDs 1224 and 1225, (m) Sequence IDs 1230 and 1231, (n) Sequence IDs 1232 and 1233, (o) Sequence IDs 1188 and 1189, (p) Sequence IDs 1190 and 1191, (q) Sequence IDs 1244 and 1245, (r) Sequence IDs 1250 and 1251-, (s) Sequence IDs 1254 and 1255, and (t) Sequence IDs 1246 and 1247 RNAi oligonucleotides selected from the group consisting of the above, according to any one of items

[51] to

[56] .

[58] The RNAi oligonucleotide according to any one of the above

[51] to

[56] , wherein the sense strand comprises SEQ ID NO: 1220 and the antisense strand comprises SEQ ID NO: 1221.

[59] The RNAi oligonucleotide according to any one of the above

[51] to

[56] , wherein the sense strand comprises SEQ ID NO: 1224 and the antisense strand comprises SEQ ID NO: 1225.

[60] The RNAi oligonucleotide according to any one of the above

[51] to

[56] , wherein the sense strand comprises SEQ ID NO: 1230 and the antisense strand comprises SEQ ID NO: 1231.

[61] The RNAi oligonucleotide according to any one of the above

[51] to

[56] , wherein the sense strand comprises SEQ ID NO: 1232 and the antisense strand comprises SEQ ID NO: 1233.

[62] The RNAi oligonucleotide according to any one of the above

[51] to

[56] , wherein the sense strand comprises sequence number 1188 and the antisense strand comprises sequence number 1189.

[63] The RNAi oligonucleotide according to any one of the above

[51] to

[56] , wherein the sense strand comprises sequence number 1190 and the antisense strand comprises sequence number 1191.

[64] The RNAi oligonucleotide according to any one of the above

[51] to

[56] , wherein the sense strand comprises SEQ ID NO: 1244 and the antisense strand comprises SEQ ID NO: 1245.

[65] The RNAi oligonucleotide according to any one of the above

[51] to

[56] , wherein the sense strand comprises SEQ ID NO: 1250 and the antisense strand comprises SEQ ID NO: 1251.

[66] The RNAi oligonucleotide according to any one of the above

[51] to

[56] , wherein the sense strand comprises SEQ ID NO: 1254 and the antisense strand comprises SEQ ID NO: 1255.

[67] The RNAi oligonucleotide according to any one of the above

[51] to

[56] , wherein the sense strand comprises SEQ ID NO: 1246 and the antisense strand comprises SEQ ID NO: 1247.

[68] A method for treating a subject having a disease, disorder, or condition related to PNPLA3 expression, comprising administering a therapeutically effective amount of an RNAi oligonucleotide or a pharmaceutical composition thereof described in any one of the preceding embodiments to the subject, thereby treating the subject.

[69] A pharmaceutical composition comprising an RNAi oligonucleotide as described in any one of the above items [1] to

[67] , and a pharmaceutically acceptable carrier, delivery agent, or excipient.

[70] A method for delivering an oligonucleotide to a target, comprising administering the pharmaceutical composition described in

[69] to the target.

[71] A method for reducing PNPLA3 expression in cells, cell populations, or subjects, the following: i. A step of contacting the cells or the cell population with an RNAi oligonucleotide described in any one of items [1] to

[67] above, or a pharmaceutical composition described in item

[69] above, ii. A step of administering to the subject an RNAi oligonucleotide described in any one of items [1] to

[67] above, or a pharmaceutical composition described in item

[69] above. The method, including the method described above.

[72] The method according to

[71] , wherein a decrease in PNPLA3 expression reduces the amount or level of PNPLA3 mRNA, the amount or level of PNPLA3 protein, or both.

[73] The method according to

[71] or

[72] , wherein the subject has a disease, disorder, or condition related to PNPLA3 expression.

[74] The method according to

[73] , wherein the disease, disorder, or condition associated with PNPLA3 expression is a cardiovascular metabolic disease, alcoholic hepatitis (AH), alcoholic liver disease (ALD), cirrhosis, hepatocellular carcinoma (HCC), non-alcoholic fatty liver disease (NAFLD), or non-alcoholic steatohepatitis (NASH).

[75] The method according to any one of the claims

[68] and

[70] to

[74] , wherein the RNAi oligonucleotide or the pharmaceutical composition is administered in combination with a second composition or therapeutic agent.

[76] A method for treating a subject having a disease, disorder, or condition related to PNPLA3 expression, comprising administering to the subject a therapeutically effective amount of an RNAi oligonucleotide comprising a sense strand and an antisense strand, wherein the sense strand and the antisense strand form a double-stranded region, the antisense strand comprises a complementary region to one of the PNPLA3 mRNA target sequences of sequence numbers 1167 to 1176, and the complementary region is at least 15 consecutive nucleotides long.

[77] A method for treating a subject having a disease, disorder, or condition related to PNPLA3 expression, comprising administering to the subject a therapeutically effective amount of an RNAi oligonucleotide or a pharmaceutical composition thereof, comprising a sense strand and an antisense strand selected from the rows listed in Table 1, Table 3, Table A, Table B, Table C, or Table D, thereby treating the subject.

[78] A method for treating a subject having a disease, disorder, or condition related to PNPLA3 expression, comprising administering to the subject a therapeutically effective amount of an RNAi oligonucleotide comprising a sense strand and an antisense strand, wherein the sense strand and the antisense strand are as follows: (a) Sequence IDs 787 and 788, (b) Sequence IDs 843 and 844, (c) Sequence IDs 867 and 868, (d) Sequence IDs 871 and 872, (e) Sequence IDs 937 and 938, (f) Sequence IDs 1003 and 1004, (g) Sequence IDs 1007 and 1008, (h) Sequence IDs 1017 and 1018, (i) Sequence IDs 1161 and 1162, and (j) Sequence IDs 1163 and 1164, (k) Sequence IDs 1220 and 1221, (l) Sequence IDs 1224 and 1225, (m) Sequence IDs 1230 and 1231, (n) Sequence IDs 1232 and 1233, (o) Sequence IDs 1188 and 1189, (p) Sequence IDs 1190 and 1191, (q) Sequence IDs 1244 and 1245, (r) Sequence IDs 1250 and 1251-, (s) Sequence IDs 1254 and 1255, and (t) Sequence IDs 1246 and 1247 The method comprising a nucleotide sequence selected from the group consisting of the following.

[79] A method for treating a subject having a disease, disorder, or condition related to PNPLA3 expression, comprising administering to the subject a therapeutically effective amount of an RNAi oligonucleotide comprising a sense strand and an antisense strand, wherein the sense strand and the antisense strand are as follows: (a) Sequence IDs 787 and 788, (b) Sequence IDs 843 and 844, (c) Sequence IDs 867 and 868, (d) Sequence IDs 871 and 872, (e) Sequence IDs 937 and 938, (f) Sequence IDs 1003 and 1004, (g) Sequence IDs 1007 and 1008, (h) Sequence IDs 1017 and 1018, (i) Sequence IDs 1161 and 1162, and (j) Sequence IDs 1163 and 1164, (k) Sequence IDs 1220 and 1221, (l) Sequence IDs 1224 and 1225, (m) Sequence IDs 1230 and 1231, (n) Sequence IDs 1232 and 1233, (o) Sequence IDs 1188 and 1189, (p) Sequence IDs 1190 and 1191, (q) Sequence IDs 1244 and 1245, (r) Sequence IDs 1250 and 1251-, (s) Sequence IDs 1254 and 1255, and (t) Sequence IDs 1246 and 1247 The method selected from the group consisting of the above.

[80] The method according to

[79] , wherein the sense strand includes sequence number 1224 and the antisense strand includes sequence number 1225, or the sense strand includes the nucleotide sequence described in sequence number 1220 and the antisense strand includes the nucleotide sequence described in sequence number 1221.

[81] The method according to

[79] , wherein the sense chain includes sequence number 1230 and the antisense chain includes sequence number 1231.

[82] The method according to

[79] , wherein the sense chain includes sequence number 1232 and the antisense chain includes sequence number 1233.

[83] The method according to

[79] , wherein the sense chain includes sequence number 1188 and the antisense chain includes sequence number 1189.

[84] The method according to

[79] , wherein the sense chain includes sequence number 1190 and the antisense chain includes sequence number 1191.

[85] The method according to

[79] , wherein the sense chain includes sequence number 1244 and the antisense chain includes sequence number 1245.

[86] The method according to

[79] , wherein the sense chain includes sequence number 1250 and the antisense chain includes sequence number 1251.

[87] The method according to

[79] , wherein the sense chain includes sequence number 1254 and the antisense chain includes sequence number 1255.

[88] The method according to

[79] , wherein the sense chain includes sequence number 1246 and the antisense chain includes sequence number 1247.

[89] The method according to any one of the above

[76] to

[88] , wherein the disease, disorder, or condition associated with PNPLA3 expression is selected from the group consisting of cardiovascular metabolic disease, alcoholic hepatitis (AH), alcoholic liver disease (ALD), cirrhosis, hepatocellular carcinoma (HCC), non-alcoholic fatty liver disease (NAFLD), and non-alcoholic steatohepatitis (NASH).

[90] Use of RNAi oligonucleotides described in any one of items [1] to

[67] above or the pharmaceutical composition described in

[69] above in the manufacture of a pharmaceutical product for the treatment of cardiovascular metabolic disease, alcoholic hepatitis (AH), alcoholic liver disease (ALD), cirrhosis, hepatocellular carcinoma (HCC), non-alcoholic fatty liver disease (NAFLD), or non-alcoholic steatohepatitis (NASH), for the treatment of a disease, disorder, or condition associated with PNPLA3 expression.

[91] RNAi oligonucleotides according to any one of items [1] to

[67] above or pharmaceutical compositions according to

[69] above, for use in the treatment of diseases, disorders, or conditions related to PNPLA3 expression, or suitable for use in the treatment of cardiovascular metabolic diseases, alcoholic hepatitis (AH), CCA, PSC, alcoholic liver disease (ALD), cirrhosis, hepatocellular carcinoma (HCC), non-alcoholic fatty liver disease (NAFLD), or non-alcoholic steatohepatitis (NASH), as selected.

[92] A kit comprising an RNAi oligonucleotide as described in any one of the preceding paragraphs [1] to

[67] , an optionally pharmaceutically acceptable carrier, and a package insert containing instructions for administration to subjects having a disease, disorder, or condition related to PNPLA3 expression.

[93] RNAi oligonucleotides or pharmaceutical compositions for or suitable for use as described in

[90] , as described in

[91] , or as suitable for use as described in

[92] , wherein the disease, disorder, or condition associated with PNPLA3 expression is a cardiovascular metabolic disease, alcoholic hepatitis (AH), alcoholic liver disease (ALD), cirrhosis, hepatocellular carcinoma (HCC), non-alcoholic fatty liver disease (NAFLD), or non-alcoholic steatohepatitis (NASH).

[0224] array The following nucleic acid sequences and / or amino acid sequences are referred to in this disclosure and are provided below for reference.

[0225] Sequence ID 1 - Wild-type human PNPLA3 (2753bp; NCBI reference sequence ID NM_025225.3)

[0226] Sequence ID 2 - Wild-type human PNPLA3 (481 aa; NCBI reference sequence ID NP_079501.2) MYDAERGWSLSFAGCGFLGFYHVGATRCLSEHAPHLLRDARMLFGASAGALHCVGVLSGIPLEQTLQVLSDLVRKARSRNIGIFHPSFNLSKFLRQGLCKCLPANVHQLISGKIGISLTR VSDGENVLVSDFRSKDEVVDALVCSCFIPFYSGLIPPSFRGVRYVDGGVSDNVPFIDAKTTITVSPFYGEYDICPKVKSTNFLHVDITKLSLRLCTGNLYLLSRAFVPPDLKVLGEICLR GYLDAFRFLEEKGICNRPQPGLKSSSEGMDPEVAMPSWANMSLDSSSPESAALAVRLEGDELLDHLRLSILPWDESILDTLSPRLATALSEEMKDKGGYMSKICNLLPIRIMSYVMLPCTL PVESAIAIVQRLVTWLPDMPDDVLWLQWVTSQVFTRVLMCLLPASRSQMPVSSQQASPCTPEQDWPCWTPCSPKGCPAETKAEATPRSILRSSLNFFLGNKVPAGAEGLSTFPSFSLEKSL

[0227] Sequence ID 3 - Human PNPLA3 I148M (Variant) mRNA

[0228] Sequence ID 4 - Human PNPLA3 I148M (Variant) MYDAERGWSLSFAGCGFLGFYHVGATRCLSEHAPHLLRDARMLFGASAGALHCVGVLSGIPLEQTLQVLSDLVRKARSRNIGIFHPSFNLSKFLRQGLCKCLPANVHQLISGKIGISLTR VSDGENVLVSDFRSKDEVVDALVCSCFMPFYSGLIPPSFRGVRYVDGGVSDNVPFIDAKTTITVSPFYGEYDICPKVKSTNFLHVDITKLSLRLCTGNLYLLSRAFVPPDLKVLGEICLR GYLDAFRFLEEKGICNRPQPGLKSSSEGMDPEVAMPSWANMSLDSSSPESAALAVRLEGDELLDHLRLSILPWDESILDTLSPRLATALSEEMKDKGGYMSKICNLLPIRIMSYVMLPCTL PVESAIAIVQRLVTWLPDMPDDVLWLQWVTSQVFTRVLMCLLPASRSQMPVSSQQASPCTPEQDWPCWTPCSPKGCPAETKAEATPRSILRSSLNFFLGNKVPAGAEGLSTFPSFSLEKSL

[0229] Sequence ID 5 - Primate PNPLA3 (2291bp; NCBI reference sequence ID XM_015457081.1)

[0230] Sequence ID 6 - Primate PNPLA3 (480 aa; NCBI reference sequence number XP_015312567.1) MYDAERGWSLSFAGCGFLGFYHVGATRCLSEHAPHLLRDARMLFGASAGALHCVGVLSGIPLEQTLQVLSDLVRKARSRNIGIFHPSFNIGKFLRQDLYKYLPANVHQLISGKICVSLTR VSDGENVLVSDFQSKDEVVDALICSCFIPFYSGLIPPSFRGVRYVDGGASDNVPFIDAKTTITVSPFYGEYDICPKVKSTNFLHVDITKLSLRLCTGNLYLLSRAFVPPDLKVLGEICLR GYLDAFRFLEEKGICNKPQRGLKSSSEGMDSEVTAPGWENTSLDSSPEPAALAMRLDGDELLDHLRLSILPWDESILDTLSPELATVSEAMKDKGGYMSKICNLLPIRIISYVMLPCTLP VESAIAIVQRLVTWLPDMPDDVQWLQWVTSQVFTRALMCLLPASRSQMPVSSEQASPCKPEQDWHCWTPCSPEDCPAEAKAEATPRSILRSSLNFFWGNKVPAGAEGLSTFPSFSLEKNL

[0231] Sequence ID 7 - Mouse PNPLA3 (4675bp; NCBI reference sequence number XM_006520346.5)

[0232] Sequence ID 8 - Mouse PNPLA3 (367 aa; NCBI reference sequence number XP_006520409.1) MEILMDLVRKARSRNIGTLHPFFNINKCIRDGLQESLPDNVHQVISGKVHISLTRVSDGENVLVSEFHSKDEVVDALVCSCFIPLFSGLIPPSFRGERYVDGGVSDNVPVLDAKTTITVSPFYGEHDICPKVKSTNFFHVNITNLSLRLCTGNLQLLTRALFPSDVKVMGELCYQGYLDAFRF LEENGICNGPQRSLSLLVAPEACLENGKLVGDKVPVSLCFTDENIWETLSPELSTALSEAIKDREGYLSKVCNLLPVRILSYIMLPCSLPVESAIAAVHRLVTWLPDIQDDIQWLQWATSQVCARMTMCLLPSTSMRFLGQSQFLGLHCIKGLKTKGKAQLSLQHASSARTTARFGAQAVQTF

[0233] Sequence ID 9-DsiRNA 1 Passenger (Sense) Strand UCUGCAGGUCCUCUCAGAUCUUGUG

[0234] Sequence ID 10-DsiRNA 1 guide (antisense) strand CACAAGAUCUGAGAGGACCUGCAGAGU

[0235] Sequence ID 11-DsiRNA 2 Passenger (Sense) Strand AGGUCCUCUCAGAUCUUGUGCGGAA

[0236] Sequence ID 12-DsiRNA 2 guide (antisense) strand UUCCGCACAAGAUCUGAGAGGACCUGC

[0237] Sequence ID 13-DsiRNA 3 Passenger (Sense) Strand AUUGGCAUCUUCCAUCCAUCCUUCA

[0238] Sequence ID 14-DsiRNA 3 guide (antisense) strand UGAAGGAUGGAUGGAAGAUGCCAAUGU

[0239] Sequence ID 15-DsiRNA 4 Passenger (Sense) Strand AUCUUCCAUCCAUCCUUCAACUUAA

[0240] Sequence ID 16-DsiRNA 4 guide (antisense) strand UUAAGUUGAAGGAUGGAUGGAUGGAAGAUGC

[0241] Sequence ID 17-DsiRNA 5 Passenger (Sense) Strand UCUUCCAUCCAUCCUUCAACUUAAG

[0242] Sequence ID 18-DsiRNA 5 guide (antisense) strand CUUAAGUUGAAGGAUGGAUGGAAGAUG

[0243] Sequence ID 19-DsiRNA 6 Passenger (Sense) Strand CCAGAGUGUCUGAUGGGGAAAACGU

[0244] Sequence ID 20-DsiRNA 6 guide (antisense) strand ACGUUUUCCCCAUCAGACACUCUGGUA

[0245] Sequence ID 21-DsiRNA 7 Passenger (Sense) Strand GAGUGUCUGAUGGGGAAAACGUUCU

[0246] Sequence ID 22-DsiRNA 7-guide (antisense) strand AGAACGUUUUCCCCAUCAGACACUCUG

[0247] Sequence ID 23-DsiRNA 8 Passenger (Sense) Strand AUGGGGAAAACGUUCUGGUGUCUGA

[0248] Sequence ID 24-DsiRNA 8 guide (antisense) strand UCAGACACCAGAACGUUUUCCCCAUCA

[0249] Sequence ID 25-DsiRNA 9 Passenger (Sense) Strand GGGGAAAACGUUCUGGUGUCUGACU

[0250] Sequence ID 26-DsiRNA 9 guide (antisense) strand AGUCAGACACCAGAACGUUUUCCCCAU

[0251] Sequence ID 27-DsiRNA 10 Passenger (Sense) Strand GGGAAAACGUUCUGGUGUCUGACUU

[0252] Sequence ID 28-DsiRNA 10 guide (antisense) strand AAGUCAGACACCAGAACGUUUUCCCCA

[0253] Sequence ID 29-DsiRNA 11 Passenger (Sense) Strand GGAAAACGUUCUGGUGUCUGACUUU

[0254] Sequence ID 30-DsiRNA 11 guide (antisense) strand AAAGUCAGACACCAGAACGUUUUCCCC

[0255] Sequence ID 31-DsiRNA 12 Passenger (Sense) Strand GAAAACGUUCUGGUGUCUGACUUUC

[0256] Sequence ID 32-DsiRNA 12-guide (antisense) strand GAAAGUCAGACACCAGAACGUUUUCCC

[0257] Sequence ID 33-DsiRNA 13 Passenger (Sense) Strand AAAACGUUCUGGUGUCUGACUUUCG

[0258] Sequence ID 34-DsiRNA 13 guide (antisense) strand CGAAAGUCAGACACCAGAACGUUUUCC

[0259] Sequence ID 35-DsiRNA 14 Passenger (Sense) Strand AAACGUUCUGGUGUCUGACUUUCGG

[0260] Sequence ID 36-DsiRNA 14 guide (antisense) strand CCGAAAGUCAGACACCAGAACGUUUUC

[0261] Sequence ID 37-DsiRNA 15 Passenger (Sense) Strand AACGUUCUGGUGUCUGACUUUCGGU

[0262] Sequence ID 38-DsiRNA 15 guide (antisense) strand ACCGAAAGUCAGACACCAGAACGUUUU

[0263] Sequence ID 39-DsiRNA 16 Passenger (Sense) Strand ACGUUCUGGUGUCUGACUUUCGGUC

[0264] Sequence ID 40-DsiRNA 16 guide (antisense) strand GACCGAAAGUCAGACACCAGAACGUUU

[0265] Sequence ID 41-DsiRNA 17 Passenger (Sense) Strand GUUCUGGUGUCUGACUUUCGGUCCA

[0266] Sequence ID 42-DsiRNA 17 guide (antisense) strand UGGACCGAAAGUCAGACACCAGAACGU

[0267] Sequence ID 43-DsiRNA 18 Passenger (Sense) Strand GACGAAGUCGUGGAUGCCUUGGUAU

[0268] Sequence ID 44-DsiRNA 18 guide (antisense) strand AUACCAAGGCAUCCACGACUUCGUCUU

[0269] Sequence ID 45-DsiRNA 19 Passenger (Sense) Strand ACGAAGUCGUGGAUGCCUUGGUAUG

[0270] Sequence ID 46-DsiRNA 19 guide (antisense) strand CAUACCAAGGCAUCCACGACUUCGUCU

[0271] Sequence ID 47-DsiRNA 20 Passenger (Sense) Strand CGAAGUCGUGGAUGCCUUGGUAUGU

[0272] Sequence ID 48-DsiRNA 20 guide (antisense) strand ACAUACCAAGGCAUCCACGACUUCGUC

[0273] Sequence ID 49-DsiRNA 21 Passenger (Sense) Strand CAGAGGCGUGCGAUAUGUGGAUGGA

[0274] Sequence ID 50-DsiRNA 21 guide (antisense) strand UCCAUCCACAUAUCGCACGCCUCUGAA

[0275] Sequence ID 51-DsiRNA 22 Passenger (Sense) Strand GCGAUAUGUGGAUGGAGGAGUGAGU

[0276] Sequence ID 52-DsiRNA 22 guide (antisense) strand ACUCACUCCUCCAUCCACAUAUCGCAC

[0277] Sequence ID 53-DsiRNA 23 Passenger (Sense) Strand GAUGGAGGAGUGAGUGACAACGUAC

[0278] Sequence ID 54-DsiRNA 23 guide (antisense) strand GUACGUUGUCACUCACUCCUCCAUCCA

[0279] Sequence ID 55-DsiRNA 24 Passenger (Sense) Strand GAGGAGUGAGUGACAACGUACCCUU

[0280] Sequence ID 56-DsiRNA 24 guide (antisense) strand AAGGGUACGUUGUCACUCACUCCUCCA

[0281] Sequence ID 57-DsiRNA 25 Passenger (Sense) Strand GAGUGAGUGACAACGUACCCUUCAU

[0282] Sequence ID 58-DsiRNA 25 guide (antisense) strand AUGAAGGGUACGUUGUCACUCUCUCCU

[0283] Sequence ID 59-DsiRNA 26 Passenger (Sense) Strand AGUGAGUGACAACGUACCCUUCAUU

[0284] Sequence ID 60-DsiRNA 26 guide (antisense) strand AAUGAAGGGUACGUUGUCACUCACUCC

[0285] Sequence ID 61-DsiRNA 27 Passenger (Sense) Strand GUGAGUGACAACGUACCCUUCAUUG

[0286] Sequence ID 62-DsiRNA 27 guide (antisense) strand CAAUGAAGGGUACGUUGUCACUCACUC

[0287] Sequence ID 63-DsiRNA 28 Passenger (Sense) Strand UGAGUGACAACGUACCCUUCAUUGA

[0288] Sequence ID 64-DsiRNA 28 guide (antisense) strand UCAAUGAAGGGUACGUUGUCACUCACU

[0289] Sequence ID 65-DsiRNA 29 Passenger (Sense) Strand GAGUGACAACGUACCCUUCAUUGAU

[0290] Sequence ID 66-DsiRNA 29 guide (antisense) strand AUCAAUGAAGGGUACGUUGUCACUCAC

[0291] Sequence ID 67-DsiRNA 30 Passenger (Sense) Strand AGUGACAACGUACCCUUCAUUGAUG

[0292] Sequence ID 68-DsiRNA 30 guide (antisense) strand CAUCAAUGAAGGGUACGUUGUCACUCA

[0293] Sequence ID 69-DsiRNA 31 Passenger (Sense) Strand GUGACAACGUACCCUUCAUUGAUGC

[0294] Sequence ID 70-DsiRNA 31 guide (antisense) strand GCAUCAAUGAAGGGUACGUUGUCACUC

[0295] Sequence ID 71-DsiRNA 32 Passenger (Sense) Strand UGACAACGUACCCUUCAUUGAUGCC

[0296] Sequence ID 72-DsiRNA 32-guide (antisense) strand GGCAUCAAUGAAGGGUACGUUGUCACU

[0297] Sequence ID 73-DsiRNA 33 Passenger (Sense) Strand GACAACGUACCCUUCAUUGAUGCCA

[0298] Sequence ID 74-DsiRNA 33 guide (antisense) strand UGGCAUCAAUGAAGGGUACGUUGUCAC

[0299] Sequence ID 75-DsiRNA 34 Passenger (Sense) Strand CAACGUACCCUUCAUUGAUGCCAAA

[0300] Sequence ID 76-DsiRNA 34 guide (antisense) strand UUUGGCAUCAAUGAAGGGUACGUUGUC

[0301] Sequence ID 77-DsiRNA 35 Passenger (Sense) Strand AACGUACCCUUCAUUGAUGCCAAAA

[0302] Sequence ID 78-DsiRNA 35 guide (antisense) strand UUUUGGCAUCAAUGAAGGGUACGUUGU

[0303] Sequence ID 79-DsiRNA 36 Passenger (Sense) Strand ACGUACCCUUCAUUGAUGCCAAAAC

[0304] Sequence ID 80-DsiRNA 36 guide (antisense) strand GUUUUGGCAUCAAUGAAGGGUACGUUG

[0305] Sequence ID 81-DsiRNA 37 Passenger (Sense) Strand CGUACCCUUCAUUGAUGCCAAAACA

[0306] Sequence ID 82-DsiRNA 37 guide (antisense) strand UGUUUUGGCAUCAAUGAAGGGUACGUU

[0307] Sequence ID 83-DsiRNA 38 Passenger (Sense) Strand UACCCUUCAUUGAUGCCAAAACAAC

[0308] Sequence ID 84-DsiRNA 38 guide (antisense) strand GUUGUUUUGGCAUCAAUGAAGGGUACG

[0309] Sequence ID 85-DsiRNA 39 Passenger (Sense) Strand UUGAUGCCAAAACAACCAUCACCGU

[0310] Sequence ID 86-DsiRNA 39 guide (antisense) strand ACGGUGAUGGUUGUUUUGGCAUCAAUG

[0311] Sequence ID 87-DsiRNA 40 Passenger (Sense) Strand GAUGCCAAAACAACCAUCACCGUGU

[0312] Sequence ID 88-DsiRNA 40 guide (antisense) strand ACACGGUGAUGGUUGUUUUGGCAUCAA

[0313] Sequence ID 89-DsiRNA 41 Passenger (Sense) Strand AUGGGGAGUACGACAUCUGCCCUAA

[0314] Sequence ID 90-DsiRNA 41 guide (antisense) strand UUAGGGCAGAUGUCGUACUCCCCAUAG

[0315] Sequence ID 91-DsiRNA 42 Passenger (Sense) Strand GUACGACAUCUGCCCUAAAGUCAAG

[0316] Sequence ID 92-DsiRNA 42 guide (antisense) strand CUUGACUUUAGGGCAGAUGUCGUACUC

[0317] Sequence ID 93-DsiRNA 43 Passenger (Sense) Strand GACAUCUGCCCUAAAGUCAAGUCCA

[0318] Sequence ID 94-DsiRNA 43 guide (antisense) strand UGGACUUGACUUUAGGGCAGAUGUCGU

[0319] Sequence ID 95-DsiRNA 44 Passenger (Sense) Strand ACAUCUGCCCUAAAGUCAAGUCCAC

[0320] Sequence ID 96-DsiRNA 44 guide (sense) strand GUGGACUUGACUUUAGGGCAGAUGUCG

[0321] Sequence ID 97-DsiRNA 45 Passenger (Sense) Strand GUCAAGUCCACGAACUUUCUUCAUG

[0322] Sequence ID 98-DsiRNA 45 guide (antisense) strand CAUGAAGAAAGUUCGUGGACUUGACUU

[0323] Sequence ID 99-DsiRNA 46 Passenger (Sense) Strand UCAAGUCCACGAACUUUCUUCAUGU

[0324] Sequence ID 100-DsiRNA 46 guide (antisense) strand ACAUGAAGAAAGUUCGUGGACUUGACU

[0325] Sequence ID 101-DsiRNA 47 Passenger (Sense) Strand CAAGUCCACGAACUUUCUUCAUGUG

[0326] Sequence ID 102-DsiRNA 47 guide (antisense) strand CACAUGAAGAAAGUUCGUGGACUUGAC

[0327] Sequence ID 103-DsiRNA 48 Passenger (Sense) Strand GUCCACGAACUUUCUUCAUGUGGAC

[0328] Sequence ID 104-DsiRNA 48 guide (antisense) strand GUCCACAUGAAGAAAGUUCGUGGACUU

[0329] Sequence ID 105-DsiRNA 49 Passenger (Sense) Strand CCACGAACUUUCUUCAUGUGGACAU

[0330] Sequence ID 106-DsiRNA 49 guide (antisense) strand AUGUCCACAUGAAGAAAGUUCGUGGAC

[0331] Sequence ID 107-DsiRNA 50 Passenger (Sense) Strand CACGAACUUUCUUCAUGUGGACAUC

[0332] Sequence ID 108-DsiRNA 50 guide (antisense) strand GAUGUCCACAUGAAGAAAGUUCGUGGA

[0333] Sequence ID 109-DsiRNA 51 Passenger (Sense) Strand ACGAACUUUCUUCAUGUGGACAUCA

[0334] Sequence ID 110-DsiRNA 51 guide (antisense) strand UGAUGUCCACAUGAAGAAAGUUCGUGG

[0335] Sequence ID 111-DsiRNA 52 Passenger (Sense) Strand CGAACUUUCUUCAUGUGGACAUCAC

[0336] Sequence ID 112-DsiRNA 52 guide (antisense) strand GUGAUGUCCACAUGAAGAAAGUUCGUG

[0337] Sequence ID 113-DsiRNA 53 Passenger (Sense) Strand GAACUUUCUUCAUGUGGACAUCACC

[0338] Sequence ID 114-DsiRNA 53 guide (antisense) strand GGUGAUGUCCACAUGAAGAAAGUUCGU

[0339] Sequence ID 115-DsiRNA 54 Passenger (Sense) Strand AACUUUCUUCAUGUGGACAUCACCA

[0340] Sequence ID 116-DsiRNA 54 guide (antisense) strand UGGUGAUGUCCACAUGAAGAAAGUUCG

[0341] Sequence ID 117-DsiRNA 55 Passenger (Sense) Strand ACUUUCUUCAUGUGGACAUCACCAA

[0342] Sequence ID 118-DsiRNA 55 guide (antisense) strand UUGGUGAUGUCCACAUGAAGAAAGUUC

[0343] Sequence ID 119-DsiRNA 56 Passenger (Sense) Strand CUUUCUUCAUGUGGACAUCACCAAG

[0344] Sequence ID 120-DsiRNA 56 guide (antisense) strand CUUGGUGAUGUCCACAUGAAGAAAGUU

[0345] Sequence ID 121-DsiRNA 57 Passenger (Sense) Strand UUUCUUCAUGUGGACAUCACCAAGC

[0346] Sequence ID 122-DsiRNA 57 guide (antisense) strand GCUUGGUGAUGUCCACAUGAAGAAAGU

[0347] Sequence ID 123-DsiRNA 58 Passenger (Sense) Strand UUCUUCAUGUGGACAUCACCAAGCU

[0348] Sequence ID 124-DsiRNA 58 guide (antisense) strand AGCUUGGUGAUGUCCACAUGAAGAAAG

[0349] Sequence ID 125-DsiRNA 59 Passenger (Sense) Strand CUUCAUGUGGACAUCACCAAGCUCA

[0350] Sequence ID 126-DsiRNA 59 guide (antisense) strand UGAGCUUGGUGAUGUCCACAUGAAGAA

[0351] Sequence ID 127-DsiRNA 60 Passenger (Sense) Strand UUCAUGUGGACAUCACCAAGCUCAG

[0352] Sequence ID 128-DsiRNA 60 guide (antisense) strand CUGAGCUUGGUGAUGUCCACAUGAAGA

[0353] Sequence ID 129-DsiRNA 61 Passenger (Sense) Strand UCAUGUGGACAUCACCAAGCUCAGU

[0354] Sequence ID 130-DsiRNA 61 guide (antisense) strand ACUGAGCUUGGUGAUGUCCACAUGAAG

[0355] Sequence ID 131-DsiRNA 62 Passenger (Sense) Strand CAUGUGGACAUCACCAAGCUCAGUC

[0356] Sequence ID 132-DsiRNA 62 guide (antisense) strand GACUGAGCUUGGUGAUGUCCACAUGAA

[0357] Sequence ID 133-DsiRNA 63 Passenger (Sense) Strand AUGUGGACAUCACCAAGCUCAGUCU

[0358] Sequence ID 134-DsiRNA 63 guide (antisense) strand AGACUGAGCUUGGUGAUGUCCACAUGA

[0359] Sequence ID 135-DsiRNA 64 Passenger (Sense) Strand UGUGGACAUCACCAAGCUCAGUCUA

[0360] Sequence ID 136-DsiRNA 64 guide (antisense) strand UAGACUGAGCUUGGUGAUGUCCACAUG

[0361] Sequence ID 137-DsiRNA 65 Passenger (Sense) Strand GUGGACAUCACCAAGCUCAGUCUAC

[0362] Sequence ID 138-DsiRNA 65 guide (antisense) strand GUAGACUGAGCUUGGUGAUGUCCACAU

[0363] Sequence ID 139-DsiRNA 66 Passenger (Sense) Strand UGGACAUCACCAAGCUCAGUCUACG

[0364] Sequence ID 140-DsiRNA 66 guide (antisense) strand CGUAGACUGAGCUUGGUGAUGUCCACA

[0365] Sequence ID 141-DsiRNA 67 Passenger (Sense) Strand AGCUUUUGUCCCCCCGGAUCUCAAG

[0366] Sequence ID 142-DsiRNA 67 guide (antisense) strand CUUGAGAUCCGGGGGGACAAAAGCUCU

[0367] Sequence ID 143-DsiRNA 68 Passenger (Sense) Strand AAGGUGCUGGGAGAGAUAUGCCUUC

[0368] Sequence ID 144-DsiRNA 68 guide (antisense) strand GAAGGCAUAUCUCUCCCAGCACCUUGA

[0369] Sequence ID 145-DsiRNA 69 Passenger (Sense) Strand GGGAGAGAUAUGCCUUCGAGGAUAU

[0370] Sequence ID 146-DsiRNA 69 guide (antisense) strand AUAUCCUCGAAGGCAUAUCUCUCCCAG

[0371] Sequence ID 147-DsiRNA 70 Passenger (Sense) Strand GGAGAGAUAUGCCUUCGAGGAUAUU

[0372] Sequence ID 148-DsiRNA 70 guide (antisense) strand AAUAUCCUCGAAGGCAUAUCUCUCCCA

[0373] Sequence ID 149-DsiRNA 71 Passenger (Sense) Strand AGAGAUAUGCCUUCGAGGAUAUUUG

[0374] Sequence ID 150-DsiRNA 71 guide (antisense) strand CAAAUAUCCUCGAAGGCAUAUCUCUCC

[0375] Sequence ID 151-DsiRNA 72 Passenger (Sense) Strand GAGAUAUGCCUUCGAGGAUAUUUGG

[0376] Sequence ID 152-DsiRNA 72-guide (antisense) strand CCAAAUAUCCUCGAAGGCAUAUCUCUC

[0377] Sequence ID 153-DsiRNA 73 Passenger (Sense) Strand AGAUAUGCCUUCGAGGAUAUUUGGA

[0378] Sequence ID 154-DsiRNA 73 guide (antisense) strand UCCAAAUAUCCUCGAAGGCAUAUCUCU

[0379] Sequence ID 155-DsiRNA 74 Passenger (Sense) Strand GAUAUGCCUUCGAGGAUAUUUGGAU

[0380] Sequence ID 156-DsiRNA 74 guide (antisense) strand AUCCAAAUAUCCUCGAAGGCAUAUCUC

[0381] Sequence ID 157-DsiRNA 75 Passenger (Sense) Strand AUAUGCCUUCGAGGAUAUUUGGAUG

[0382] Sequence ID 158-DsiRNA 75 guide (antisense) strand CAUCCAAAUAUCCUCGAAGGCAUAUCU

[0383] Sequence ID 159-DsiRNA 76 passenger (sense) strand AUGCCUUCGAGGAUAUUUGGAUGCA

[0384] Sequence ID 160-DsiRNA 76 guide (antisense) strand UGCAUCCAAAUAUCCUCGAAGGCAUAU

[0385] Sequence ID 161-DsiRNA 77 Passenger (Sense) Strand UGCCUUCGAGGAUAUUUGGAUGCAU

[0386] Sequence ID 162-DsiRNA 77 guide (antisense) strand AUGCAUCCAAAUAUCCUCGAAGGCAUA

[0387] Sequence ID 163-DsiRNA 78 Passenger (Sense) Strand GCCUUCGAGGAUAUUUGGAUGCAUU

[0388] Sequence ID 164-DsiRNA 78 guide (antisense) strand AAUGCAUCCAAAUAUCCUCGAAGGCAU

[0389] Sequence ID 165-DsiRNA 79 Passenger (Sense) Strand UCAGGUUCUUGGAAGAGAAGGGCAU

[0390] Sequence ID 166-DsiRNA 79 guide (antisense) strand AUGCCCUUCUCUUCCAAGAACCUGAAU

[0391] Sequence ID 167-DsiRNA 80 Passenger (Sense) Strand CAGGUUCUUGGAAGAGAAGGGCAUC

[0392] Sequence ID 168-DsiRNA 80 guide (antisense) strand GAUGCCCUUCUCUUCCAAGAACCUGAA

[0393] Sequence ID 169-DsiRNA 81 Passenger (Sense) Strand AGGUUCUUGGAAGAGAAGGGCAUCU

[0394] Sequence ID 170-DsiRNA 81 guide (antisense) strand AGAUGCCCUUCUCUUCCAAGAACCUGA

[0395] Sequence ID 171-DsiRNA 82 Passenger (Sense) Strand GGUUCUUGGAAGAGAAGGGCAUCUG

[0396] Sequence ID 172-DsiRNA 82 guide (antisense) strand CAGAUGCCCUUCUCUUCCAAGAACCUG

[0397] Sequence ID 173-DsiRNA 83 Passenger (Sense) Strand UGGAAGAGAAGGGCAUCUGCAACAG

[0398] Sequence ID 174-DsiRNA 83 guide (antisense) strand CUGUUGCAGAUGCCCUUCUCUUCCAAG

[0399] Sequence ID 175-DsiRNA 84 Passenger (Sense) Strand UGAAGUCAUCCUCAGAAGGGAUGGA

[0400] Sequence ID 176-DsiRNA 84 guide (antisense) strand UCCAUCCCUUCUGAGGAUGACUUCAGG

[0401] Sequence ID 177-DsiRNA 85 Passenger (Sense) Strand GAAGUCAUCCUCAGAAGGGAUGGAU

[0402] Sequence ID 178-DsiRNA 85 guide (antisense) strand AUCCAUCCCUUCUGAGGAUGACUUCAG

[0403] Sequence ID 179-DsiRNA 86 Passenger (Sense) Strand AAGUCAUCCUCAGAAGGGAUGGAUC

[0404] Sequence ID 180-DsiRNA 86 guide (antisense) strand GAUCCAUCCCUUCUGAGGAUGACUUCA

[0405] Sequence ID 181-DsiRNA 87 Passenger (Sense) Strand GUCAUCCUCAGAAGGGAUGGAUCCU

[0406] Sequence ID 182-DsiRNA 87 guide (antisense) strand AGGAUCCAUCCCUUCUGAGGAUGACUU

[0407] Sequence ID 183-DsiRNA 88 Passenger (Sense) Strand CAUCCUCAGAAGGGAUGGAUCCUGA

[0408] Sequence ID 184-DsiRNA 88 guide (antisense) strand UCAGGAUCCAUCCCUUCUGAGGAUGAC

[0409] Sequence ID 185-DsiRNA 89 Passenger (Sense) Strand AUCCUCAGAAGGGAUGGAUCCUGAG

[0410] Sequence ID 186 (DsiRNA 89 guide (antisense) strand) CUCAGGAUCCAUCCCUUCUGAGGAUGA

[0411] Sequence ID 187-DsiRNA 90 Passenger (Sense) Strand CUAGACCACCUGCGUCUCAGCAUCC

[0412] Sequence ID 188-DsiRNA 90 guide (antisense) strand GGAUGCUGAGACGCAGGUGGUCUAGCA

[0413] Sequence ID 189-DsiRNA 91 Passenger (Sense) Strand AGUGAAGAAAUGAAAGACAAAGGUG

[0414] Sequence ID 190-DsiRNA 91 guide (antisense) strand CACCUUUGUCUUUCAUUUCUUCACUCA

[0415] Sequence ID 191-DsiRNA 92 Passenger (Sense) Strand GUGAAGAAAUGAAAGACAAAGGUGG

[0416] Sequence ID 192-DsiRNA 92 guide (antisense) strand CCACCUUUGUCUUUCAUUUCUUCACUC

[0417] Sequence ID 193-DsiRNA 93 Passenger (Sense) Strand UGAAGAAAUGAAAGACAAAGGUGGA

[0418] Sequence ID 194-DsiRNA 93 guide (antisense) strand UCCACCUUUGUCUUUCAUUUCUUCACU

[0419] Sequence ID 195-DsiRNA 94 Passenger (Sense) Strand GAAGAAAUGAAAGACAAAGGUGGAU

[0420] Sequence ID 196-DsiRNA 94 guide (antisense) strand AUCCACCUUUGUCUUUCAUUUCUUCAC

[0421] Sequence ID 197-DsiRNA 95 Passenger (Sense) Strand AAGAAAUGAAAGACAAAGGUGGAUA

[0422] Sequence ID 198-DsiRNA 95 guide (antisense) strand UAUCCACCUUUGUCUUUCAUUUCUUCA

[0423] Sequence ID 199-DsiRNA 96 Passenger (Sense) Strand AGAAAUGAAAGACAAAGGUGGAUAC

[0424] Sequence ID 200-DsiRNA 96 guide (antisense) strand GUAUCCACCUUUGUCUUUCAUUUCUUC

[0425] Sequence ID 201-DsiRNA 97 Passenger (Sense) Strand GAAAUGAAAGACAAAGGUGGAUACA

[0426] Sequence ID 202-DsiRNA 97 guide (antisense) strand UGUAUCCACCUUUGUCUUUCAUUUCUU

[0427] Sequence ID 203-DsiRNA 98 Passenger (Sense) Strand AAAUGAAAGACAAAGGUGGAUACAU

[0428] Sequence ID 204-DsiRNA 98 guide (antisense) strand AUGUAUCCACCUUUGUCUUUCAUUUCU

[0429] Sequence ID 205-DsiRNA 99 Passenger (Sense) Strand AAUGAAAGACAAAGGUGGAUACAUG

[0430] Sequence ID 206-DsiRNA 99 guide (antisense) strand CAUGUAUCCACCUUUGUCUUUCAUUUC

[0431] Sequence ID 207-DsiRNA 100 Passenger (Sense) Strand AUGAAAGACAAAGGUGGAUACAUGA

[0432] Sequence ID 208-DsiRNA 100 guide (antisense) strand UCAUGUAUCCACCUUUGUCUUUCAUUU

[0433] Sequence ID 209-DsiRNA 101 Passenger (Sense) Strand UGAAAGACAAAGGUGGAUACAUGAG

[0434] Sequence ID 210-DsiRNA 101 guide (antisense) strand CUCAUGUAUCCACCUUUGUCUUUCAUU

[0435] Sequence ID 211-DsiRNA 102 Passenger (Sense) Strand GAAAGACAAAGGUGGAUACAUGAGC

[0436] Sequence ID 212-DsiRNA 102 guide (antisense) strand GCUCAUGUAUCCACCUUUGUCUUUCAU

[0437] Sequence ID 213-DsiRNA 103 Passenger (Sense) Strand AAGACAAAGGUGGAUACAUGAGCAA

[0438] Sequence ID 214-DsiRNA 103 guide (antisense) strand UUGCUCAUGUAUCCACCUUUGUCUUUC

[0439] Sequence ID 215-DsiRNA 104 Passenger (Sense) Strand AGACAAAGGUGGAUACAUGAGCAAG

[0440] Sequence ID 216-DsiRNA 104 guide (antisense) strand CUUGCUCAUGUAUCCACCUUUGUCUUU

[0441] Sequence ID 217-DsiRNA 105 Passenger (Sense) Strand GACAAAGGUGGAUACAUGAGCAAGA

[0442] Sequence ID 218-DsiRNA 105 guide (antisense) strand UCUUGCUCAUGUAUCCACCUUUGUCUU

[0443] Sequence ID 219-DsiRNA 106 Passenger (Sense) Strand ACAAAGGUGGAUACAUGAGCAAGAU

[0444] Sequence ID 220-DsiRNA 106 guide (antisense) strand AUCUUGCUCAUGUAUCCACCUUUGUCU

[0445] Sequence ID 221-DsiRNA 107 Passenger (Sense) Strand AAGGUGGAUACAUGAGCAAGAUUUG

[0446] Sequence ID 222-DsiRNA 107 guide (antisense) strand CAAAUCUUGCUCAUGUAUCCACCUUUG

[0447] Sequence ID 223-DsiRNA 108 Passenger (Sense) Strand AGGUGGAUACAUGAGCAAGAUUUGC

[0448] Sequence ID 224-DsiRNA 108 guide (antisense) strand GCAAAUCUUGCUCAUGUAUCCACCUUU

[0449] Sequence ID 225-DsiRNA 109 Passenger (Sense) Strand UGGAUACAUGAGCAAGAUUUGCAAC

[0450] Sequence ID 226-DsiRNA 109 guide (antisense) strand GUUGCAAAUCUUGCUCAUGUAUCCACC

[0451] Sequence ID 227-DsiRNA 110 Passenger (Sense) Strand GGAUACAUGAGCAAGAUUUGCAACU

[0452] Sequence ID 228-DsiRNA 110 guide (antisense) strand AGUUGCAAAUCUUGCUCAUGUAUCCAC

[0453] Sequence ID 229-DsiRNA 111 Passenger (Sense) Strand GAUACAUGAGCAAGAUUUGCAACUU

[0454] Sequence ID 230 - DsiRNA 111 guide (antisense) strand AAGUUGCAAAUCUUGCUCAUGUAUCCA

[0455] Sequence ID 231-DsiRNA 112 Passenger (Sense) Strand AUACAUGAGCAAGAUUUGCAACUUG

[0456] Sequence ID 232-DsiRNA 112 guide (antisense) strand CAAGUUGCAAAUCUUGCUCAUGUAUCC

[0457] Sequence ID 233-DsiRNA 113 Passenger (Sense) Strand UACAUGAGCAAGAUUUGCAACUUGC

[0458] Sequence ID 234-DsiRNA 113 guide (antisense) strand GCAAGUUGCAAAUCUUGCUCAUGUAUC

[0459] Sequence ID 235-DsiRNA 114 Passenger (Sense) Strand ACAUGAGCAAGAUUUGCAACUUGCU

[0460] Sequence ID 236-DsiRNA 114 guide (antisense) strand AGCAAGUUGCAAAUCUUGCUCAUGUAU

[0461] Sequence ID 237-DsiRNA 115 Passenger (Sense) Strand CAUGAGCAAGAUUUGCAACUUGCUA

[0462] Sequence ID 238-DsiRNA 115 guide (antisense) strand UAGCAAGUUGCAAAUCUUGCUCAUGUA

[0463] Sequence ID 239: DsiRNA 116 Passenger (Sense) Strand AUGAGCAAGAUUUGCAACUUGCUAC

[0464] Sequence ID 240-DsiRNA 116 guide (antisense) strand GUAGCAAGUUGCAAAUCUUGCUCAUGU

[0465] Sequence ID 241-DsiRNA 117 Passenger (Sense) Strand UGAGCAAGAUUUGCAACUUGCUACC

[0466] Sequence ID 242-DsiRNA 117 guide (antisense) strand GGUAGCAAGUUGCAAAUCUUGCUCAUG

[0467] Sequence ID 243-DsiRNA 118 Passenger (Sense) Strand GAGCAAGAUUUGCAACUUGCUACCC

[0468] Sequence ID 244-DsiRNA 118 guide (antisense) strand GGGUAGCAAGUUGCAAAUCUUGCUCAU

[0469] Sequence ID 245-DsiRNA 119 Passenger (Sense) Strand AGCAAGAUUUGCAACUUGCUACCCA

[0470] Sequence ID 246-DsiRNA 119 guide (antisense) strand UGGGUAGCAAGUUGCAAAUCUUGCUCA

[0471] Sequence ID 247-DsiRNA 120 Passenger (Sense) Strand GCAAGAUUUGCAACUUGCUACCCAU

[0472] Sequence ID 248-DsiRNA 120 guide (antisense) strand AUGGGUAGCAAGUUGCAAAUCUUGCUC

[0473] Sequence ID 249-DsiRNA 121 Passenger (Sense) Strand CAAGAUUUGCAACUUGCUACCCAUU

[0474] Sequence ID 250-DsiRNA 121 guide (antisense) strand AAUGGGUAGCAAGUUGCAAAUCUUGCU

[0475] Sequence ID 251-DsiRNA 122 Passenger (Sense) Strand AAGAUUUGCAACUUGCUACCCAUUA

[0476] Sequence ID 252-DsiRNA 122 guide (antisense) strand UAAUGGGUAGCAAGUUGCAAAUCUUGC

[0477] Sequence ID 253-DsiRNA 123 Passenger (Sense) Strand AGAUUUGCAACUUGCUACCCAUUAG

[0478] Sequence ID 254-DsiRNA 123 guide (antisense) strand CUAAUGGGUAGCAAGUUGCAAAUCUUG

[0479] Sequence ID 255-DsiRNA 124 Passenger (Sense) Strand GAUUUGCAACUUGCUACCCAUUAGG

[0480] Sequence ID 256-DsiRNA 124 guide (antisense) strand CCUAAUGGGUAGCAAGUUGCAAAUCUU

[0481] Sequence ID 257-DsiRNA 125 Passenger (Sense) Strand AUUUGCAACUUGCUACCCAUUAGGA

[0482] Sequence ID 258-DsiRNA 125 guide (antisense) strand UCCUAAUGGGUAGCAAGUUGCAAAUCU

[0483] Sequence ID 259-DsiRNA 126 Passenger (Sense) Strand UUUGCAACUUGCUACCCAUUAGGAU

[0484] Sequence ID 260-DsiRNA 126 guide (antisense) strand AUCCUAAUGGGUAGCAAGUUGCAAAUC

[0485] Sequence ID 261-DsiRNA 127 Passenger (Sense) Strand UUGCAACUUGCUACCCAUUAGGAUA

[0486] Sequence ID 262-DsiRNA 127 guide (antisense) strand UAUCCUAAUGGGUAGCAAGUUGCAAAU

[0487] Sequence ID 263-DsiRNA 128 Passenger (Sense) Strand UGCAACUUGCUACCCAUUAGGAUAA

[0488] Sequence ID 264-DsiRNA 128 guide (antisense) strand UUAUCCUAAUGGGUAGCAAGUUGCAAA

[0489] Sequence ID 265-DsiRNA 129-Passenger (Sense) Strand GCAACUUGCUACCCAUUAGGAUAAU

[0490] Sequence ID 266-DsiRNA 129 guide (antisense) strand AUUAUCCUAAUGGGUAGCAAGUUGCAA

[0491] Sequence ID 267-DsiRNA 130 Passenger (Sense) Strand CAACUUGCUACCCAUUAGGAUAAUG

[0492] Sequence ID 268-DsiRNA 130 guide (antisense) strand CAUUAUCCUAAUGGGUAGCAAGUUGCA

[0493] Sequence ID 269-DsiRNA 131 Passenger (Sense) Strand CUUGCUACCCAUUAGGAUAAUGUCU

[0494] Sequence ID 270 - DsiRNA 131 guide (antisense) strand AGACAUUAUCCUAAUGGGUAGCAAGUU

[0495] Sequence ID 271-DsiRNA 132 Passenger (Sense) Strand UUGCUACCCAUUAGGAUAAUGUCUU

[0496] Sequence ID 272-DsiRNA 132 guide (antisense) strand AAGACAUUAUCCUAAUGGGUAGCAAGU

[0497] Sequence ID 273-DsiRNA 133 Passenger (Sense) Strand UGCUACCCAUUAGGAUAAUGUCUUA

[0498] Sequence ID 274-DsiRNA 133 guide (antisense) strand UAAGACAUUAUCCUAAUGGGUAGCAAG

[0499] Sequence ID 275-DsiRNA 134 Passenger (Sense) Strand AUUAGGAUAAUGUCUUAUGUAAUGC

[0500] Sequence ID 276-DsiRNA 134 guide (antisense) strand GCAUUACAUAAGACAUUAUCCUAAUGG

[0501] Sequence ID 277-DsiRNA 135 Passenger (Sense) Strand UUAGGAUAAUGUCUUAUGUAAUGCU

[0502] Sequence ID 278-DsiRNA 135 guide (antisense) strand AGCAUUACAUAAGACAUUAUCCUAAUG

[0503] Sequence ID 279-DsiRNA 136 Passenger (Sense) Strand CUGUGGAAUCUGCCAUUGCGAUUGU

[0504] Sequence ID 280-DsiRNA 136 guide (antisense) strand ACAAUCGCAAUGGCAGAUUCCACAGGC

[0505] Sequence ID 281-DsiRNA 137 Passenger (Sense) Strand GGAAUCUGCCAUUGCGAUUGUCCAG

[0506] Sequence ID 282-DsiRNA 137 guide (antisense) strand CUGGACAAUCGCAAUGGCAGAUUCCAC

[0507] Sequence ID 283-DsiRNA 138 Passenger (Sense) Strand GAAUCUGCCAUUGCGAUUGUCCAGA

[0508] Sequence ID 284-DsiRNA 138 guide (antisense) strand UCUGGACAAUCGCAAUGGCAGAUUCCA

[0509] Sequence ID 285-DsiRNA 139 Passenger (Sense) Strand AAUCUGCCAUUGCGAUUGUCCAGAG

[0510] Sequence ID 286-DsiRNA 139 guide (antisense) strand CUCUGGACAAUCGCAAUGGCAGAUUCC

[0511] Sequence ID 287-DsiRNA 140 Passenger (Sense) Strand AUCUGCCAUUGCGAUUGUCCAGAGA

[0512] Sequence ID 288-DsiRNA 140 guide (antisense) strand UCUCUGGACAAUCGCAAUGGCAGAUUC

[0513] Sequence ID 289-DsiRNA 141 Passenger (Sense) Strand CCAUUGCGAUUGUCCAGAGACUGGU

[0514] Sequence ID 290-DsiRNA 141 guide (antisense) strand ACCAGUCUCUGGACAAUCGCAAUGGCA

[0515] Sequence ID 291-DsiRNA 142 Passenger (Sense) Strand CAUUGCGAUUGUCCAGAGACUGGUG

[0516] Sequence ID 292-DsiRNA 142 guide (antisense) strand CACCAGUCUCUGGACAAUCGCAAUGGC

[0517] Sequence ID 293-DsiRNA 143 Passenger (Sense) Strand AUUGCGAUUGUCCAGAGACUGGUGA

[0518] Sequence ID 294-DsiRNA 143 guide (antisense) strand UCACCAGUCUCUGGACAAUCGCAAUGG

[0519] Sequence ID 295-DsiRNA 144 Passenger (Sense) Strand UUGCGAUUGUCCAGAGACUGGUGAC

[0520] Sequence ID 296-DsiRNA 144 guide (antisense) strand GUCACCAGUCUCUGGACAAUCGCAAUG

[0521] Sequence ID 297-DsiRNA 145 Passenger (Sense) Strand UGCGAUUGUCCAGAGACUGGUGACA

[0522] Sequence ID 298-DsiRNA 145 guide (antisense) strand UGUCACCAGUCUCUGGACAAUCGCAAU

[0523] Sequence ID 299-DsiRNA 146 Passenger (Sense) Strand GCGAUUGUCCAGAGACUGGUGACAU

[0524] Sequence ID 300-DsiRNA 146 guide (antisense) strand AUGUCACCAGUCUCUGGACAAUCGCAA

[0525] Sequence ID 301-DsiRNA 147 Passenger (Sense) Strand CGAUUGUCCAGAGACUGGUGACAUG

[0526] Sequence ID 302-DsiRNA 147 guide (antisense) strand CAUGUCACCAGUCUCUGGACAAUCGCA

[0527] Sequence ID 303-DsiRNA 148 Passenger (Sense) AUUGUCCAGAGACUGGUGACAUGGC

[0528] Sequence ID 304-DsiRNA 148 guide (antisense) strand GCCAUGUCACCAGUCUCUGGACAAUCG

[0529] Sequence ID 305-DsiRNA 149 Passenger (Sense) Strand GUCCAGAGACUGGUGACAUGGCUUC

[0530] Sequence ID 306-DsiRNA 149 guide (antisense) strand GAAGCCAUGUCACCAGUCUCUGGACAA

[0531] Sequence ID 307-DsiRNA 150 Passenger (Sense) Strand CCAGAGACUGGUGACAUGGCUUCCA

[0532] Sequence ID 308-DsiRNA 150 guide (antisense) strand UGGAAGCCAUGUCACCAGUCUCUGGAC

[0533] Sequence ID 309-DsiRNA 151 Passenger (Sense) CAGAGACUGGUGACAUGGCUUCCAG

[0534] Sequence ID 310-DsiRNA 151 guide (antisense) strand CUGGAAGCCAUGUCACCAGUCUCUGGA

[0535] Sequence ID 311-DsiRNA 152 Passenger (Sense) Strand AGAGACUGGUGACAUGGCUUCCAGA

[0536] Sequence ID 312-DsiRNA 152 guide (antisense) strand UCUGGAAGCCAUGUCACCAGUCUCUGG

[0537] Sequence ID 313-DsiRNA 153 Passenger (Sense) Strand GAGACUGGUGACAUGGCUUCCAGAU

[0538] Sequence ID 314-DsiRNA 153 guide (antisense) strand AUCUGGAAGCCAUGUCACCAGUCUCUG

[0539] Sequence ID 315-DsiRNA 154 Passenger (Sense) Strand AGACUGGUGACAUGGCUUCCAGAUA

[0540] Sequence ID 316-DsiRNA 154 guide (antisense) strand UAUCUGGAAGCCAUGUCACCAGUCUCU

[0541] Sequence ID 317-DsiRNA 155 Passenger (Sense) Strand GACUGGUGACAUGGCUUCCAGAUAU

[0542] Sequence ID 318-DsiRNA 155 guide (antisense) strand AUAUCUGGAAGCCAUGUCACCAGUCUC

[0543] Sequence ID 319-DsiRNA 156 Passenger (Sense) Strand ACUGGUGACAUGGCUUCCAGAUAUG

[0544] Sequence ID 320-DsiRNA 156 guide (antisense) strand CAUAUCUGGAAGCCAUGUCACCAGUCU

[0545] Sequence ID 321-DsiRNA 157 Passenger (Sense) Strand CUGGUGACAUGGCUUCCAGAUAUGC

[0546] Sequence ID 322-DsiRNA 157 guide (antisense) strand GCAUAUCUGGAAGCCAUGUCACCAGUC

[0547] Sequence ID 323-DsiRNA 158 Passenger (Sense) Strand GACAUGGCUUCCAGAUAUGCCCGAC

[0548] Sequence ID 324-DsiRNA 158 guide (antisense) strand GUCGGGCAUAUCUGGAAGCCAUGUCAC

[0549] Sequence ID 325-DsiRNA 159 Passenger (Sense) Strand CAUGGCUUCCAGAUAUGCCCGACGA

[0550] Sequence ID 326-DsiRNA 159 guide (antisense) strand UCGUCGGGCAUAUCUGGAAGCCAUGUC

[0551] Sequence ID 327-DsiRNA 160 Passenger (Sense) Strand CAGAUAUGCCCGACGAUGUCCUGUG

[0552] Sequence ID 328-DsiRNA 160 guide (antisense) strand CACAGGACAUCGUCGGGCAUAUCUGGA

[0553] Sequence ID 329-DsiRNA 161 Passenger (Sense) Strand CUCACAGGUGUUCACUCGAGUGCUG

[0554] Sequence ID 330-DsiRNA 161 guide (antisense) strand CAGCACUCGAGUGAACACCUGUGAGGU

[0555] Sequence ID 331-DsiRNA 162 Passenger (Sense) Strand CACUCGAGUGCUGAUGUGUCUGCUC

[0556] Sequence ID 332-DsiRNA 162 guide (antisense) strand GAGCAGACACAUCAGCACUCGAGUGAA

[0557] Sequence ID 333-DsiRNA 163 Passenger (Sense) Strand CUCGAGUGCUGAUGUGUCUGCUCCC

[0558] Sequence ID 334-DsiRNA 163 guide (antisense) strand GGGAGCAGACACAUCAGCACUCGAGUG

[0559] Sequence ID 335-DsiRNA 164 Passenger (Sense) Strand CCCAAAUGCCAGUGAGCAGCCAACA

[0560] Sequence ID 336-DsiRNA 164 guide (antisense) strand UGUUGGCUGCUCACUGGCAUUUGGGAC

[0561] Sequence ID 337-DsiRNA 165 Passenger (Sense) Strand UCAGGUCCAGCCUGAACUUCUUCUU

[0562] Sequence ID 338-DsiRNA 165 guide (antisense) strand AAGAAGAAGUUCAGGCUGGACCUGAGG

[0563] Sequence ID 339-DsiRNA 166 Passenger (Sense) Strand CAGGUCCAGCCUGAACUUCUUCUUG

[0564] Sequence ID 340-DsiRNA 166 guide (antisense) strand CAAGAAGAAGUUCAGGCUGGACCUGAG

[0565] Sequence ID 341-DsiRNA 167 Passenger (Sense) Strand AGGUCCAGCCUGAACUUCUUCUUGG

[0566] Sequence ID 342-DsiRNA 167 guide (antisense) strand CCAAGAAGAAGUUCAGGCUGGACCUGA

[0567] Sequence ID 343-DsiRNA 168 Passenger (Sense) Strand CAAUAAAGUACCUGCUGGUGCUGAG

[0568] Sequence ID 344-DsiRNA 168 guide (antisense) strand CUCAGCACCAGCAGGUACUUUAUUGCC

[0569] Sequence ID 345-DsiRNA 169 Passenger (Sense) Strand AAUAAAGUACCUGCUGGUGCUGAGG

[0570] Sequence ID 346-DsiRNA 169 guide (antisense) strand CCUCAGCACCAGCAGGUACUUUAUUGC

[0571] Sequence ID 347-DsiRNA 170 Passenger (Sense) Strand AUAAAGUACCUGCUGGUGCUGAGGG

[0572] Sequence ID 348-DsiRNA 170 guide (antisense) strand CCCUCAGCACCAGCAGGUACUUUAUUG

[0573] Sequence ID 349-DsiRNA 171 Passenger (Sense) Strand CUCUCCACCUUUCCCAGUUUUUCAC

[0574] Sequence ID 350-DsiRNA 171 guide (antisense) strand GUGAAAAACUGGGAAAGGUGGAGAGCC

[0575] Sequence ID 351-DsiRNA 172 Passenger (Sense) Strand CUCCACCUUUCCCAGUUUUUCACUA

[0576] Sequence ID 352-DsiRNA 172 guide (antisense) strand UAGUGAAAAACUGGGAAAGGUGGAGAG

[0577] Sequence ID 353-DsiRNA 173 Passenger (Sense) Strand UCCACCUUUCCCAGUUUUUCACUAG

[0578] Sequence ID 354-DsiRNA 173 guide (antisense) strand CUAGUGAAAAACUGGGAAAGGUGGAGA

[0579] Sequence ID 355-DsiRNA 174 Passenger (Sense) Strand CCACCUUUCCCAGUUUUUCACUAGA

[0580] Sequence ID 356-DsiRNA 174 guide (antisense) strand UCUAGUGAAAAACUGGGAAAGGUGGAG

[0581] Sequence ID 357-DsiRNA 175 Passenger (Sense) Strand CACCUUUCCCAGUUUUUCACUAGAG

[0582] Sequence ID 358-DsiRNA 175 guide (antisense) strand CUCUAGUGAAAAACUGGGAAAGGUGGA

[0583] Sequence ID 359-DsiRNA 176 Passenger (Sense) Strand ACCUUUCCCAGUUUUUCACUAGAGA

[0584] Sequence ID 360-DsiRNA 176 guide (antisense) strand UCUCUAGUGAAAAACUGGGAAAGGUGG

[0585] Sequence ID 361-DsiRNA 177 Passenger (Sense) Strand CCUUUCCCAGUUUUUCACUAGAGAA

[0586] Sequence ID 362-DsiRNA 177 guide (antisense) strand UUCUCUAGUGAAAAACUGGGAAAGGUG

[0587] Sequence ID 363-DsiRNA 178 Passenger (Sense) Strand AGUUUUUCACUAGAGAAGAGUCUGU

[0588] Sequence ID 364-DsiRNA 178 guide (antisense) strand ACAGACUCUUCUCUAGUGAAAAACUGG

[0589] Sequence ID 365-DsiRNA 179 Passenger (Sense) Strand UCUAGCAGAUUCUUUCAGAGGUGCU

[0590] Sequence ID 366-DsiRNA 179 guide (antisense) strand AGCACCUCUGAAAGAAUCUGCUAGACU

[0591] Sequence ID 367-DsiRNA 180 Passenger (Sense) Strand AGCAGAUUCUUUCAGAGGUGCUAAA

[0592] Sequence ID 368-DsiRNA 180 guide (antisense) strand UUUAGCACCUCUGAAAGAAUCUGCUAG

[0593] Sequence ID 369-DsiRNA 181 Passenger (Sense) Strand GCAGAUUCUUUCAGAGGUGCUAAAG

[0594] Sequence ID 370-DsiRNA 181 guide (antisense) strand CUUUAGCACCUCUGAAAGAAUCUGCUA

[0595] Sequence ID 371-DsiRNA 182 Passenger (Sense) Strand CAGAUUCUUUCAGAGGUGCUAAAGU

[0596] Sequence ID 372-DsiRNA 182 guide (antisense) strand ACUUUAGCACCUCUGAAAGAAUCUGCU

[0597] Sequence ID 373-DsiRNA 183 Passenger (Sense) Strand AGAUUCUUUCAGAGGUGCUAAAGUU

[0598] Sequence ID 374-DsiRNA 183 guide (antisense) strand AACUUUAGCACCUCUGAAAGAAUCUGC

[0599] Sequence ID 375-DsiRNA 184 Passenger (Sense) Strand GAUUCUUUCAGAGGUGCUAAAGUUU

[0600] Sequence ID 376-DsiRNA 184 guide (antisense) strand AAACUUUAGCACCUCUGAAAGAAUCUG

[0601] Sequence ID 377-DsiRNA 185 Passenger (Sense) Strand AUUCUUUCAGAGGUGCUAAAGUUUC

[0602] Sequence ID 378-DsiRNA 185 guide (antisense) strand GAAACUUUAGCACCUCUGAAAGAAUCU

[0603] Sequence ID 379-DsiRNA 186 Passenger (Sense) Strand AGGUGCUAAAGUUUCCCAUCUUUGU

[0604] Sequence ID 380-DsiRNA 186 guide (antisense) strand ACAAAGAUGGGAAACUUUAGCACCUCU

[0605] Sequence ID 381-DsiRNA 187 Passenger (Sense) Strand GGUGCUAAAGUUUCCCAUCUUUGUG

[0606] Sequence ID 382-DsiRNA 187 guide (antisense) strand CACAAAGAUGGGAAACUUUAGCACCUC

[0607] Sequence ID 383-DsiRNA 188 Passenger (Sense) Strand GUGCUAAAGUUUCCCAUCUUUGUGC

[0608] Sequence ID 384-DsiRNA 188 guide (antisense) strand GCACAAAGAUGGGAAACUUUAGCACCU

[0609] Sequence ID 385-DsiRNA 189 Passenger (Sense) Strand GCUAAAGUUUCCCAUCUUUGUGCAG

[0610] Sequence ID 386-DsiRNA 189 guide (antisense) strand CUGCACAAAGAUGGGAAACUUUAGCAC

[0611] Sequence ID 387-DsiRNA 190 Passenger (Sense) Strand CUAAAGUUUCCCAUCUUUGUGCAGC

[0612] Sequence ID 388-DsiRNA 190 guide (antisense) strand GCUGCACAAAGAUGGGAAACUUUAGCA

[0613] Sequence ID 389-DsiRNA 191 Passenger (Sense) Strand UAAAGUUUCCCAUCUUUGUGCAGCU

[0614] Sequence ID 390-DsiRNA 191 guide (antisense) strand AGCUGCACAAAGAUGGGAAACUUUAGC

[0615] Sequence ID 391-DsiRNA 192 Passenger (Sense) Strand AAAGUUUCCCAUCUUUGUGCAGCUA

[0616] Sequence ID 392-DsiRNA 192 guide (antisense) strand UAGCUGCACAAAGAUGGGAAACUUUAG

[0617] Sequence ID 393-DsiRNA 193 Passenger (Sense) Strand AAGUUUCCCAUCUUUGUGCAGCUAC

[0618] Sequence ID 394-DsiRNA 193 guide (antisense) strand GUAGCUGCACAAAGAUGGGAAACUUUA

[0619] Sequence ID 395-DsiRNA 194 Passenger (Sense) Strand AGUUUCCCAUCUUUGUGCAGCUACC

[0620] Sequence ID 396-DsiRNA 194 guide (antisense) strand GGUAGCUGCACAAAGAUGGGAAACUUU

[0621] Sequence ID 397-DsiRNA 195 Passenger (Sense) Strand GUUUCCCAUCUUUGUGCAGCUACCU

[0622] Sequence ID 398-DsiRNA 195 guide (antisense) strand AGGUAGCUGCACAAAGAUGGGAAACUU

[0623] Sequence ID 399-DsiRNA 196 Passenger (Sense) Strand AUCUUUGUGCAGCUACCUCCGCAUU

[0624] Sequence ID 400-DsiRNA 196 guide (antisense) strand AAUGCGGAGGUAGCUGCACAAAGAUGG

[0625] Sequence ID 401-DsiRNA 197 Passenger (Sense) Strand GUGCAGCUACCUCCGCAUUGCUGUG

[0626] Sequence ID 402-DsiRNA 197 guide (antisense) strand CACAGCAAUGCGGAGGUAGCUGCACAA

[0627] Sequence ID 403-DsiRNA 198 Passenger (Sense) Strand CCAGCCUCUGAGCUGAGUUGGUUUU

[0628] Sequence ID 404-DsiRNA 198 guide (antisense) strand AAAACCAACUCAGCUCAGAGGCUGGGA

[0629] Sequence ID 405-DsiRNA 199 Passenger (Sense) Strand CAGCCUCUGAGCUGAGUUGGUUUUA

[0630] Sequence ID 406-DsiRNA 199 guide (antisense) strand UAAAACCAACUCAGCUCAGAGGCUGGG

[0631] Sequence ID 407-DsiRNA 200 Passenger (Sense) Strand AGCCUCUGAGCUGAGUUGGUUUUAU

[0632] Sequence ID 408-DsiRNA 200 guide (antisense) strand AUAAAACCAACUCAGCUCAGAGGCUGG

[0633] Sequence ID 409-DsiRNA 201 Passenger (Sense) Strand GCCUCUGAGCUGAGUUGGUUUUAUG

[0634] Sequence ID 410-DsiRNA 201 guide (antisense) strand CAUAAAACCAACUCAGCUCAGAGGCUG

[0635] Sequence ID 411-DsiRNA 202 Passenger (Sense) Strand CCUCUGAGCUGAGUUGGUUUUAUGA

[0636] Sequence ID 412-DsiRNA 202 guide (antisense) strand UCAUAAAACCAACUCAGCUCAGAGGCU

[0637] Sequence ID 413-DsiRNA 203 Passenger (Sense) Strand CUCUGAGCUGAGUUGGUUUUAUGAA

[0638] Sequence ID 414-DsiRNA 203 guide (antisense) strand UUCAUAAAACCAACUCAGCUCAGAGGC

[0639] Sequence ID 415-DsiRNA 204 Passenger (Sense) Strand UCUGAGCUGAGUUGGUUUUAUGAAA

[0640] Sequence ID 416-DsiRNA 204 guide (antisense) strand UUUCAUAAAACCAACUCAGCUCAGAGG

[0641] Sequence ID 417-DsiRNA 205 Passenger (Sense) Strand UGAGCUGAGUUGGUUUUAUGAAAAG

[0642] Sequence ID 418-DsiRNA 205 guide (antisense) strand CUUUUCAUAAAACCAACUCAGCUCAGA

[0643] Sequence ID 419-DsiRNA 206 Passenger (Sense) Strand UGAGUUGGUUUUAUGAAAAGCUAGG

[0644] Sequence ID 420-DsiRNA 206 guide (antisense) strand CCUAGCUUUUCAUAAAACCAACUCAGC

[0645] Sequence ID 421-DsiRNA 207 Passenger (Sense) Strand GAGUUGGUUUUAUGAAAAGCUAGGA

[0646] Sequence ID 422-DsiRNA 207 guide (antisense) strand UCCUAGCUUUUCAUAAAACCAACUCAG

[0647] Sequence ID 423-DsiRNA 208 Passenger (Sense) Strand UUGGUUUUAUGAAAAGCUAGGAAGC

[0648] Sequence ID 424-DsiRNA 208 guide (antisense) strand GCUUCCUAGCUUUUCAUAAAACCAACU

[0649] Sequence ID 425-DsiRNA 209 Passenger (Sense) Strand UGGUUUUAUGAAAAGCUAGGAAGCA

[0650] Sequence ID 426-DsiRNA 209 guide (antisense) strand UGCUUCCUAGCUUUUCAUAAAACCAAC

[0651] Sequence ID 427-DsiRNA 210 Passenger (Sense) Strand GGUUUUAUGAAAAGCUAGGAAGCAA

[0652] Sequence ID 428-DsiRNA 210 guide (antisense) strand UUGCUUCCUAGCUUUUCAUAAAACCAA

[0653] Sequence ID 429-DsiRNA 211 Passenger (Sense) Strand UUUUAUGAAAAGCUAGGAAGCAACC

[0654] Sequence ID 430-DsiRNA 211 guide (antisense) strand GGUUGCUUCCUAGCUUUUCAUAAAACC

[0655] Sequence ID 431-DsiRNA 212 Passenger (Sense) Strand UUAUGAAAAGCUAGGAAGCAACCUU

[0656] Sequence ID 432-DsiRNA 212 guide (antisense) strand AAGGUUGCUUCCUAGCUUUUCAUAAAA

[0657] Sequence ID 433-DsiRNA 213 Passenger (Sense) Strand UAUGAAAAGCUAGGAAGCAACCUUU

[0658] Sequence ID 434-DsiRNA 213 guide (antisense) strand AAAGGUUGCUUCCUAGCUUUUCAUAAA

[0659] Sequence ID 435-DsiRNA 214 Passenger (Sense) Strand AUGAAAAGCUAGGAAGCAACCUUUC

[0660] Sequence ID 436-DsiRNA 214 guide (antisense) strand GAAAGGUUGCUUCCUAGCUUUUCAUAA

[0661] Sequence ID 437-DsiRNA 215 Passenger (Sense) Strand CCAGCACUUAACUCUAAUACAUCAG

[0662] Sequence ID 438-DsiRNA 215 guide (antisense) strand CUGAUGUAUUAGAGUUAAGUGCUGGAC

[0663] Sequence ID 439-DsiRNA 216 Passenger (Sense) Strand CAGCACUUAACUCUAAUACAUCAGC

[0664] Sequence ID 440-DsiRNA 216 guide (antisense) strand GCUGAUGUAUUAGAGUUAAGUGCUGGA

[0665] Sequence ID 441-DsiRNA 217 Passenger (Sense) Strand UAAUACAUCAGCAUGCGUUAAUUCA

[0666] Sequence ID 442-DsiRNA 217 guide (antisense) strand UGAAUUAACGCAUGCUGAUGUAUUAGA

[0667] Sequence ID 443-DsiRNA 218 Passenger (Sense) Strand AAUACAUCAGCAUGCGUUAAUUCAG

[0668] Sequence ID 444-DsiRNA 218 guide (antisense) strand CUGAAUUAACGCAUGCUGAUGUAUUAG

[0669] Sequence ID 445-DsiRNA 219 Passenger (Sense) Strand AGCAUGCGUUAAUUCAGCUGGUUGG

[0670] Sequence ID 446-DsiRNA 219 guide (antisense) strand CCAACCAGCUGAAUUAACGCAUGCUGA

[0671] Sequence ID 447-DsiRNA 220 Passenger (Sense) Strand GCAUGCGUUAAUUCAGCUGGUUGGG

[0672] Sequence ID 448-DsiRNA 220 guide (antisense) strand CCCAACCAGCUGA AUUAACGCAUGCUG

[0673] Sequence ID 449-DsiRNA 221 Passenger (Sense) Strand UGCGUUAAUUCAGCUGGUUGGGAAA

[0674] Sequence ID 450-DsiRNA 221 guide (antisense) strand UUUCCCAACCAGCUGAAUUAACGCAUG

[0675] Sequence ID 451-DsiRNA 222 Passenger (Sense) Strand GCGUUAAUUCAGCUGGUUGGGAAAU

[0676] Sequence ID 452-DsiRNA 222 guide (antisense) strand AUUUCCCAACCAGCUGAAUUAACGCAU

[0677] Sequence ID 453-DsiRNA 223 Passenger (Sense) Strand CGUUAAUUCAGCUGGUUGGGAAAUG

[0678] Sequence ID 454-DsiRNA 223 guide (antisense) strand CAUUUCCCAACCAGCUGAAUUAACGCA

[0679] Sequence ID 455-DsiRNA 224 Passenger (Sense) Strand GUUAAUUCAGCUGGUUGGGAAAUGA

[0680] Sequence ID 456-DsiRNA 224 guide (antisense) strand UCAUUUCCCAACCAGCUGAAUUAACGC

[0681] Sequence ID 457-DsiRNA 225 Passenger (Sense) Strand UUAAUUCAGCUGGUUGGGAAAUGAC

[0682] Sequence ID 458-DsiRNA 225 guide (antisense) strand GUCAUUUCCCAACCAGCUGAAUUAACG

[0683] Sequence ID 459-DsiRNA 226 Passenger (Sense) Strand UAAUUCAGCUGGUUGGGAAAUGACA

[0684] Sequence ID 460-DsiRNA 226 guide (antisense) strand UGUCAUUUCCCAACCAGCUGAAUUAAC

[0685] Sequence ID 461-DsiRNA 227 Passenger (Sense) Strand AUUCAGCUGGUUGGGAAAUGACACC

[0686] Sequence ID 462-DsiRNA 227 guide (antisense) strand GGUGUCAUUUCCCAACCAGCUGAAUUA

[0687] Sequence ID 463-DsiRNA 228 Passenger (Sense) Strand UUCAGCUGGUUGGGAAAUGACACCA

[0688] Sequence ID 464-DsiRNA 228 guide (antisense) strand UGGUGUCAUUUCCCAACCAGCUGAAUU

[0689] Sequence ID 465-DsiRNA 229 Passenger (Sense) Strand UCAGCUGGUUGGGAAAUGACACCAG

[0690] Sequence ID 466-DsiRNA 229 guide (antisense) strand CUGGUGUCAUUUCCCAACCAGCUGAAU

[0691] Sequence ID 467-DsiRNA 230 Passenger (Sense) Strand CAGCUGGUUGGGAAAUGACACCAGG

[0692] Sequence ID 468-DsiRNA 230 guide (antisense) strand CCUGGUGUCAUUUCCCAACCAGCUGAA

[0693] Sequence ID 469-DsiRNA 231 Passenger (Sense) Strand AGCUGGUUGGGAAAUGACACCAGGA

[0694] Sequence ID 470-DsiRNA 231 guide (antisense) strand UCCUGGUGUCAUUUCCCAACCAGCUGA

[0695] Sequence ID 471-DsiRNA 232 Passenger (Sense) Strand GCUGGUUGGGAAAUGACACCAGGAA

[0696] Sequence ID 472-DsiRNA 232 guide (antisense) strand UUCCUGGUGUCAUUUCCCAACCAGCUG

[0697] Sequence ID 473-DsiRNA 233 Passenger (Sense) Strand GCAGAGGGUCCCUUACUGACUGUUU

[0698] Sequence ID 474-DsiRNA 233 guide (antisense) strand AAACAGUCAGUAAGGGACCCUCUGCAC

[0699] Sequence ID 475-DsiRNA 234 Passenger (Sense) Strand CAGAGGGUCCCUUACUGACUGUUUC

[0700] Sequence ID 476-DsiRNA 234 guide (antisense) strand GAAACAGUCAGUAAGGGACCCUCUGCA

[0701] Sequence ID 477-DsiRNA 235 Passenger (Sense) Strand AGAGGGUCCCUUACUGACUGUUUCG

[0702] Sequence ID 478-DsiRNA 235 guide (antisense) strand CGAAACAGUCAGUAAGGGACCCUCUGC

[0703] Sequence ID 479-DsiRNA 236 Passenger (Sense) Strand CCUAUUAAUGGUCAGACUGUUCCAG

[0704] Sequence ID 480-DsiRNA 236 guide (antisense) strand CUGGAACAGUCUGACCAUUAAUAGGGC

[0705] Sequence ID 481-DsiRNA 237 Passenger (Sense) Strand CUAUUAAUGGUCAGACUGUUCCAGC

[0706] Sequence ID 482-DsiRNA 237 guide (antisense) strand GCUGGAACAGUCUGACCAUUAAUAGGG

[0707] Sequence ID 483-DsiRNA 238 Passenger (Sense) Strand UAUUAAUGGUCAGACUGUUCCAGCA

[0708] Sequence ID 484-DsiRNA 238 guide (antisense) strand UGCUGGAACAGUCUGACCAUUAAUAGG

[0709] Sequence ID 485-DsiRNA 239 Passenger (Sense) Strand AUUAAUGGUCAGACUGUUCCAGCAU

[0710] Sequence ID 486-DsiRNA 239 guide (antisense) strand AUGCUGGAACAGUCUGACCAUUAAUAG

[0711] Sequence ID 487-DsiRNA 240 Passenger (Sense) Strand UUAAUGGUCAGACUGUUCCAGCAUG

[0712] Sequence ID 488-DsiRNA 240 guide (antisense) strand CAUGCUGGAACAGUCUGACCAUUAAUA

[0713] Sequence ID 489-DsiRNA 241 Passenger (Sense) Strand UAAUGGUCAGACUGUUCCAGCAUGA

[0714] Sequence ID 490-DsiRNA 241 guide (antisense) strand UCAUGCUGGAACAGUCUGACCAUUAAU

[0715] Sequence ID 491-DsiRNA 242 Passenger (Sense) Strand AAUGGUCAGACUGUUCCAGCAUGAG

[0716] Sequence ID 492-DsiRNA 242 guide (antisense) strand CUCAUGCUGGAACAGUCUGACCAUUAA

[0717] Sequence ID 493-DsiRNA 243 Passenger (Sense) Strand AGAACGACACUGCCUGUCAGGUGGU

[0718] Sequence ID 494-D iSCSI 243 guide (antisense) strand ACCACCUGACAGGCAGUGUCGUUCUUG

[0719] Sequence ID 495-DsiRNA 244 Passenger (Sense) Strand CGACACUGCCUGUCAGGUGGUCUGC

[0720] Sequence ID 496-DsiRNA 244 guide (antisense) strand GCAGACCACCUGACAGGCAGUGUCGUU

[0721] Sequence ID 497-DsiRNA 245 Passenger (Sense) Strand AACCUUGACUACUAAAAACGUCUCC

[0722] Sequence ID 498-DsiRNA 245 guide (antisense) strand GGAGACGUUUUUAGUAGUCAAGGUUAU

[0723] Sequence ID 499-DsiRNA 246 Passenger (Sense) Strand UUUAGAACACCUUUUUCACCUAACU

[0724] Sequence ID 500-DsiRNA 246 guide (antisense) strand AGUUAGGUGAAAAAGGUGUUCUAAAAU

[0725] Sequence ID 501-DsiRNA 247 Passenger (Sense) Strand UUAGAACACCUUUUUCACCUAACUA

[0726] Sequence ID 502-DsiRNA 247 guide (antisense) strand UAGUUAGGUGAAAAAGGUGUUCUAAAA

[0727] Sequence ID 503-DsiRNA 248 Passenger (Sense) Strand UAGAACACCUUUUUCACCUAACUAA

[0728] Sequence ID 504-DsiRNA 248 guide (antisense) strand UUAGUUAGGUGAAAAAGGUGUUCUAAA

[0729] Sequence ID 505-DsiRNA 249 Passenger (Sense) Strand AGAACACCUUUUUCACCUAACUAAA

[0730] Sequence ID 506-DsiRNA 249 guide (antisense) strand UUUAGUUAGGUGAAAAAGGUGUUCUAA

[0731] Sequence ID 507-DsiRNA 250 Passenger (Sense) Strand GAACACCUUUUUCACCUAACUAAAA

[0732] Sequence ID 508-DsiRNA 250 guide (antisense) strand UUUUAGUUAGGUGAAAAAGGUGUUCUA

[0733] Sequence ID 509-DsiRNA 251 Passenger (Sense) Strand AACACCUUUUUCACCUAACUAAAAU

[0734] Sequence ID 510-DsiRNA 251 guide (antisense) strand AUUUUAGUUAGGUGAAAAAGGUGUUCU

[0735] Sequence ID 511-DsiRNA 252 Passenger (Sense) Strand ACACCUUUUUCACCUAACUAAAAUA

[0736] Sequence ID 512-DsiRNA 252 guide (antisense) strand UAUUUUAGUUAGGUGAAAAAGGUGUUC

[0737] Sequence ID 513-DsiRNA 253 Passenger (Sense) Strand CACCUUUUUCACCUAACUAAAAUAA

[0738] Sequence ID 514-DsiRNA 253 guide (antisense) strand UUAUUUUAGUUAGGUGAAAAAGGUGUU

[0739] Sequence ID 515-DsiRNA 254 Passenger (Sense) Strand ACCUUUUUCACCUAACUAAAAUAAU

[0740] Sequence ID 516-DsiRNA 254 guide (antisense) strand AUUAUUUUAGUUAGGUGAAAAAGGUGU

[0741] Sequence ID 517-DsiRNA 255 Passenger (Sense) Strand CUUUUUCACCUAACUAAAAUAAUGU

[0742] Sequence ID 518-DsiRNA 255 guide (antisense) strand ACAUUAUUUUAGUUAGGUGAAAAAGGU

[0743] Sequence ID 519-DsiRNA 256 Passenger (Sense) Strand UUUUCACCUAACUAAAAUAAUGUUU

[0744] Sequence ID 520-DsiRNA 256 guide (antisense) strand AAACAUUAUUUUAGUUAGGUGAAAAAG

[0745] Sequence ID 521-DsiRNA 257 Passenger (Sense) Strand UUCACCUAACUAAAAUAAUGUUUAA

[0746] Sequence ID 522-DsiRNA 257 guide (antisense) strand UUAAACAUUAUUUUAGUUAGGUGAAAA

[0747] Sequence ID 523-DsiRNA 258 Passenger (Sense) Strand UCACCUAACUAAAAUAAUGUUUAAA

[0748] Sequence ID 524-DsiRNA 258 guide (antisense) strand UUUAAACAUUAUUUUAGUUADGUGAAA

[0749] Sequence ID 525-DsiRNA 259 Passenger (Sense) Strand CACCUAACUAAAAUAAUGUUUAAAG

[0750] Sequence ID 526-DsiRNA 259 guide (antisense) strand CUUUAAACAUUAUUUUAGUUAGGUGAA

[0751] Sequence ID 527-DsiRNA 260 Passenger (Sense) Strand ACCUAACUAAAAUAAUGUUUAAAGA

[0752] Sequence ID 528-DsiRNA 260 guide (antisense) strand UCUUUAAACAUUAUUUUAGUUAGGUGA

[0753] Sequence ID 529-DsiRNA 261 Passenger (Sense) Strand CCUAACUAAAAUAAUGUUUAAAGAG

[0754] Sequence ID 530-DsiRNA 261 guide (antisense) strand CUCUUUAAACAUUAUUUUAGUUAGGUG

[0755] Sequence ID 531-DsiRNA 262 Passenger (Sense) Strand CUAACUAAAAUAAUGUUUAAAGAGU

[0756] Sequence ID 532-DsiRNA 262 guide (antisense) strand ACUCUUUAAACAUUAUUUUAGUUAGGU

[0757] Sequence ID 533-DsiRNA 263 Passenger (Sense) Strand UAACUAAAAUAAUGUUUAAAGAGUU

[0758] Sequence ID 534-DsiRNA 263 guide (antisense) strand AACUCUUUAAACAUUAUUUUAGUUAGG

[0759] Sequence ID 535-DsiRNA 264 Passenger (Sense) Strand AACUAAAAUAAUGUUUAAAGAGUUU

[0760] Sequence ID 536-DsiRNA 264 guide (antisense) strand AAACUCUUUAAACAUUAUUUUAGUU Organ

[0761] Sequence ID 537-DsiRNA 265 Passenger (Sense) Strand ACUAAAAUAAUGUUUAAAGAGUUUU

[0762] Sequence ID 538-DsiRNA 265 guide (antisense) strand AAAACUCUUUAAACAUUAUUUUAGUUA

[0763] Sequence ID 539-DsiRNA 266 passenger (sense) strand CUAAAAUAAUGUUUAAAGAGUUUUG

[0764] Sequence ID 540-DsiRNA 266 guide (antisense) strand CAAAACUCUUUAAACAUUAUUUUAGUU

[0765] Sequence ID 541-DsiRNA 267 Passenger (Sense) Strand UAAAAUAAUGUUUAAAGAGUUUUGU

[0766] Sequence ID 542-DsiRNA 267 guide (antisense) strand ACAAAACUCUUUAAACAUUAUUUUAGU

[0767] Sequence ID 543-DsiRNA 268 Passenger (Sense) Strand AAGAGUUUUGUAUAAAAAUGUAAGG

[0768] Sequence ID 544-DsiRNA 268 guide (antisense) strand CCUUACAUUUUUAUACAAAACUCUUUA

[0769] Sequence ID 545-DsiRNA 269 Passenger (Sense) Strand GUUUUGUAUAAAAAUGUAAGGAAGC

[0770] Sequence ID 546-DsiRNA 269 guide (antisense) strand GCUUCCUUACAUUUUUAUACAAAACUC

[0771] Sequence ID 547-DsiRNA 270 Passenger (Sense) Strand UUUUGUAUAAAAAUGUAAGGAAGCG

[0772] Sequence ID 548-DsiRNA 270 guide (antisense) strand CGCUUCCUUACAUUUUUAUACAAAACU

[0773] Sequence ID 549-DsiRNA 271 Passenger (Sense) Strand UUUGUAUAAAAAUGUAAGGAAGCGU

[0774] Sequence ID 550-DsiRNA 271 guide (antisense) strand ACGCUUCCUUACAUUUUUAUACAAAAC

[0775] Sequence ID 551-DsiRNA 272 Passenger (Sense) Strand UUGUAUAAAAAUGUAAGGAAGCGUU

[0776] Sequence ID 552-DsiRNA 272 guide (antisense) strand AACGCUUCCUUACAUUUUUAUACAAAA

[0777] Sequence ID 553-DsiRNA 273 Passenger (Sense) Strand UGUAUAAAAAUGUAAGGAAGCGUUG

[0778] Sequence ID 554-DsiRNA 273 guide (antisense) strand CAACGCUUCCUUACAUUUUUAUACAAA

[0779] Sequence ID 555-DsiRNA 274 Passenger (Sense) Strand GUAUAAAAAUGUAAGGAAGCGUUGU

[0780] Sequence ID 556-DsiRNA 274 guide (antisense) strand ACAACGCUUCCUUACAUUUUUAUACAA

[0781] Sequence ID 557-DsiRNA 275 Passenger (Sense) Strand AUGUAAGGAAGCGUUGUUACCUGUU

[0782] Sequence ID 558-DsiRNA 275 guide (antisense) strand AACAGGUAACAACGCUUCCUUACAUUU

[0783] Sequence ID 559-DsiRNA 276 Passenger (Sense) Strand UUUUGUAUUAUGUGAAUCAGUGAGA

[0784] Sequence ID 560-DsiRNA 276 guide (antisense) strand UCUCACUGAUUCACAUAAUACAAAAUU

[0785] Sequence ID 561-DsiRNA 277 Passenger (Sense) Strand UUUGUAUUAUGUGAAUCAGUGAGAU

[0786] Sequence ID 562-DsiRNA 277 guide (antisense) strand AUCUCACUGAUUCACAUAAUACAAAAU

[0787] Sequence ID 563-DsiRNA 278 Passenger (Sense) Strand UUGUAUUAUGUGAAUCAGUGAGAUG

[0788] Sequence ID 564-DsiRNA 278 guide (antisense) strand CAUCUCACUGAUUCACAUAAUACAAAA

[0789] Sequence ID 565-DsiRNA 279 Passenger (Sense) Strand UGUAUUAUGUGAAUCAGUGAGAUGU

[0790] Sequence ID 566-DsiRNA 279 guide (antisense) strand ACAUCUCACUGAUUCACAUAAUACAAA

[0791] Sequence ID 567-DsiRNA 280 Passenger (Sense) Strand GUAUUAUGUGAAUCAGUGAGAUGUU

[0792] Sequence ID 568-DsiRNA 280 guide (antisense) strand AACAUCUCACUGAUUCACAUAAUACAA

[0793] Sequence ID 569-DsiRNA 281 Passenger (Sense) Strand UAUUAUGUGAAUCAGUGAGAUGUUA

[0794] Sequence ID 570-DsiRNA 281 guide (antisense) strand UAACAUCUCACUGAUUCACAUAAUACA

[0795] Sequence ID 571-DsiRNA 282 Passenger (Sense) Strand AUUAUGUGAAUCAGUGAGAUGUUAG

[0796] Sequence ID 572-DsiRNA 282 guide (antisense) strand CUAACAUCUCACUGAUUCACAUAAUAC

[0797] Sequence ID 573-DsiRNA 283 Passenger (Sense) Strand UUAUGUGAAUCAGUGAGAUGUUAGU

[0798] Sequence ID 574-DsiRNA 283 guide (antisense) strand ACUAACAUCUCACUGAUUCACAUAAUA

[0799] Sequence ID 575-DsiRNA 284 Passenger (Sense) Strand UAUGUGAAUCAGUGAGAUGUUAGUA

[0800] Sequence ID 576-DsiRNA 284 guide (antisense) strand UACUAACAUCUCACUGAUUCACAUAAU

[0801] Sequence ID 577-DsiRNA 285 Passenger (Sense) Strand AUGUGAAUCAGUGAGAUGUUAGUAG

[0802] Sequence ID 578-DsiRNA 285 guide (antisense) strand CUACUAACAUCUCACUGAUUCACAUAA

[0803] Sequence ID 579-DsiRNA 286 Passenger (Sense) Strand UGUGAAUCAGUGAGAUGUUAGUAGA

[0804] Sequence ID 580-DsiRNA 286 guide (antisense) strand UCUACUAACAUCUCACUGAUUCACAUA

[0805] Sequence ID 581-DsiRNA 287 Passenger (Sense) Strand GUGAAUCAGUGAGAUGUUAGUAGAA

[0806] Sequence ID 582-DsiRNA 287 guide (antisense) strand UUCUACUAACAUCUCACUGAUUCACAU

[0807] Sequence ID 583-DsiRNA 288 Passenger (Sense) Strand GUGAGAUGUUAGUAGAAUAAGCCUU

[0808] Sequence ID 584-DsiRNA 288 guide (antisense) strand AAGGCUUAUUCUACUAACAUCUCACUG

[0809] Sequence ID 585-DsiRNA 289 Passenger (Sense) Strand UUUUCUAUUUAUGCAUUUGAGUACA

[0810] Sequence ID 586-DsiRNA 289 guide (antisense) strand UGUACUCAAAUGCAUAAAUAGAAAAAA

[0811] Sequence ID 587-DsiRNA 290 Passenger (Sense) Strand UUUCUAUUUAUGCAUUUGAGUACAG

[0812] Sequence ID 588-DsiRNA 290 guide (antisense) strand CUGUACUCAAAUGCAUAAAUAGAAAAA

[0813] Sequence ID 589-DsiRNA 291 Passenger (Sense) Strand UUCUAUUUAUGCAUUUGAGUACAGT

[0814] Sequence ID 590-DsiRNA 291 guide (antisense) strand ACUGUACUCAAAUGCAUAAAUAGAAAA

[0815] Sequence ID 591-DsiRNA 292 Passenger (Sense) Strand CUAUUUAUGCAUUUGAGUACAGUAC

[0816] Sequence ID 592-DsiRNA 292 guide (antisense) strand GUACUGUACUCAAAUGCAUAAAUAGAA

[0817] Sequence ID 593-DsiRNA 293 Passenger (Sense) Strand UGCUCAAACUGUUAAAUGUUGGAAA

[0818] Sequence ID 594-DsiRNA 293 guide (antisense) strand UUUCCAACAUUUAACAGUUUGAGCACA

[0819] Sequence ID-DsiRNA 294 Passenger (Sense) Strand GCUCAAACUGUUAAAUGUUGGAAAA595

[0820] Sequence ID 596-DsiRNA 294 guide (antisense) strand UUUUCCAACAUUUAACAGUUUGAGCAC

[0821] Sequence ID 597-DsiRNA 295 Passenger (Sense) Strand CUCAAACUGUUAAAUGUUGGAAAAG

[0822] Sequence ID 598-DsiRNA 295 guide (antisense) strand CUUUUCCAACAUUUAACAGUUUGAGCA

[0823] Sequence ID 599-DsiRNA 296 Passenger (Sense) Strand UCAAACUGUUAAAUGUUGGAAAAGA

[0824] Sequence ID 600-DsiRNA 296 guide (antisense) strand UCUUUUCCAACAUUUAACAGUUUGAGC

[0825] Sequence ID 601-DsiRNA 297 Passenger (Sense) Strand CAAACUGUUAAAUGUUGGAAAAGAA

[0826] Sequence ID 602-DsiRNA 297 guide (antisense) strand UUCUUUUCCAACAUUUAACAGUUUGAG

[0827] Sequence ID 603-DsiRNA 298 Passenger (Sense) Strand AAACUGUUAAAUGUUGGAAAAGAAA

[0828] Sequence ID 604-DsiRNA 298 guide (antisense) strand UUUCUUUUCCAACAUUUAACAGUUUGA

[0829] Sequence ID 605-DsiRNA 299 Passenger (Sense) Strand AACUGUUAAAUGUUGGAAAAGAAAG

[0830] Sequence ID 606-DsiRNA 299 guide (antisense) strand CUUUCUUUUCCAACAUUUAACAGUUUG

[0831] Sequence ID 607-DsiRNA 300 Passenger (Sense) Strand ACUGUUAAAUGUUGGAAAAGAAAGA

[0832] Sequence ID 608-DsiRNA 300 guide (antisense) strand UCUUUCUUUUCCAACAUUUAACAGUUU

[0833] Sequence ID 609-DsiRNA 301 Passenger (Sense) Strand CUGUUAAAUGUUGGAAAAGAAAGAT

[0834] Sequence ID 610-DsiRNA 301 guide (antisense) strand AUCUUUCUUUUCCAACAUUUAACAGUU

[0835] Sequence ID 611-DsiRNA 302 Passenger (Sense) Strand UGUUAAAUGUUGGAAAAGAAAGATA

[0836] Sequence ID 612-DsiRNA 302 guide (antisense) strand UAUCUUUCUUUUCCAACAUUUAACAGU

[0837] Sequence ID 613-DsiRNA 303 Passenger (Sense) Strand GUUAAAUGUUGGAAAAGAAAGAUAC

[0838] Sequence ID 614-DsiRNA 303 guide (antisense) strand GUAUCUUUCUUUUCCAACAUUUAACAG

[0839] Sequence ID 615-DsiRNA 304 Passenger (Sense) Strand UUAAAUGUUGGAAAAGAAAGAUACA

[0840] Sequence ID 616-DsiRNA 304 guide (antisense) strand UGUAUCUUUCUUUUCCAACAUUUAACA

[0841] Sequence ID 617-DsiRNA 305 Passenger (Sense) Strand UAAAUGUUGGAAAAGAAAGAUACAA

[0842] Sequence ID 618-DsiRNA 305 guide (antisense) strand UUGUAUCUUUCUUUUCCAACAUUUAAC

[0843] Sequence ID 619-DsiRNA 306 Passenger (Sense) Strand GCACUUGACUGAGAAGACAGACCCT

[0844] Sequence ID 620-DsiRNA 306 guide (antisense) strand AGGGUCUGUCUUCUCUCAGUCAAGUGCUU

[0845] Sequence ID 621-DsiRNA 307 Passenger (Sense) Strand GAGAAAAGAGGCUACUUGUGAAAAT

[0846] Sequence ID 622-DsiRNA 307 guide (antisense) strand AUUUUCACAAGUAGCCUCUUUUCUCAA

[0847] Sequence ID 623-DsiRNA 308 Passenger (Sense) Strand AGAAAAGAGGCUACUUGUGAAAATA

[0848] Sequence ID 624-DsiRNA 308 guide (antisense) strand UAUUUUCACAAGUAGCCUCUUUUCUCA

[0849] Sequence ID 625-DsiRNA 309 Passenger (Sense) Strand GAAAAGAGGCUACUUGUGAAAAUAA

[0850] Sequence ID 626-DsiRNA 309 guide (antisense) strand UUAUUUUCACAAGUAGCCUCUUUUCUC

[0851] Sequence ID 627-DsiRNA 310 Passenger (Sense) Strand AAAAGAGGCUACUUGUGAAAAUAAT

[0852] Sequence ID 628-DsiRNA 310 guide (antisense) strand AUUAUUUUCACAAGUAGCCUCUUUUCU

[0853] Sequence ID 629-DsiRNA 311 Passenger (Sense) Strand AAAGAGGCUACUUGUGAAAAUAATG

[0854] Sequence ID 630-DsiRNA 311 guide (antisense) strand CAUUAUUUUCACAAGUAGCCUCUUUUC

[0855] Sequence ID 631-DsiRNA 312 Passenger (Sense) Strand AAGAGGCUACUUGUGAAAAUAAUGA

[0856] Sequence ID 632-DsiRNA 312 guide (antisense) strand UCAUUAUUUUCACAAGUAGCCUCUUUU

[0857] Sequence ID 633-DsiRNA 313 Passenger (Sense) Strand AGAGGCUACUUGUGAAAAUAAUGAG

[0858] Sequence ID 634-DsiRNA 313 guide (antisense) strand CUCAUUAUUUUCACAAGUAGCCUCUUU

[0859] Sequence ID 635-DsiRNA 314 Passenger (Sense) Strand GAGGCUACUUGUGAAAAUAAUGAGC

[0860] Sequence ID 636-DsiRNA 314 guide (antisense) strand GCUCAUUAUUUUCACAAGUAGCCUCUU

[0861] Sequence ID 637-DsiRNA 315 Passenger (Sense) Strand GGCUACUUGUGAAAAUAAUGAGCCC

[0862] Sequence ID 638-DsiRNA 315 guide (antisense) strand GGGCUCAUUAUUUUCACAAGUAGCCUC

[0863] Sequence ID 639-DsiRNA 316 Passenger (Sense) Strand CUACUUGUGAAAAUAAUGAGCCCCC

[0864] Sequence ID 640-DsiRNA 316 guide (antisense) strand GGGGGCUCAUUAUUUUCACAAGUAGCC

[0865] Sequence ID 641-DsiRNA 317 Passenger (Sense) Strand UGAACCUGCCUUCUUACAUCUUGAG

[0866] Sequence ID 642-DsiRNA 317 guide (antisense) strand CUCAAGAUGUAAGAAGGCAGGUUCAAA

[0867] Sequence ID 643-DsiRNA 318 Passenger (Sense) Strand AAAGUUACAAGUUUCUUUUCCCAAG

[0868] Sequence ID 644-DsiRNA 318 guide (antisense) strand CUUGGGAAAAGAAACUUGUAACUUUCC

[0869] Sequence ID 645-DsiRNA 319 Passenger (Sense) Strand AAGUUACAAGUUUCUUUUCCCAAGT

[0870] Sequence ID 646-DsiRNA 319 guide (antisense) strand ACUUGGGAAAAGAAACUUGUAACUUUC

[0871] Sequence ID 647-DsiRNA 320 Passenger (Sense) Strand AGUUACAAGUUUCUUUUCCCAAGTT

[0872] Sequence ID 648-DsiRNA 320 guide (antisense) strand AACUUGGGAAAAGAAACUUGUAACUUU

[0873] Sequence ID 649-DsiRNA 321 Passenger (Sense) Strand AGUUUCUUUUCCCAAGUUUCCCAGT

[0874] Sequence ID 650-DsiRNA 321 guide (antisense) strand ACUGGGAAACUUGGGAAAAGAAACUUG

[0875] Sequence ID 651-DsiRNA 322 Passenger (Sense) Strand CAACAGUAUUUUCUAAUAACCAGTA

[0876] Sequence ID 652-DsiRNA 322 guide (antisense) strand UACUGGUUAUUAGAAAAUACUGUUGGC

[0877] Sequence ID 653-DsiRNA 323 Passenger (Sense) Strand AACAGUAUUUUCUAAUAACCAGUAT

[0878] Sequence ID 654-DsiRNA 323 guide (antisense) strand AUACUGGUUAUUAGAAAAUACUGUUGG

[0879] Sequence ID 655-DsiRNA 324 Passenger (Sense) Strand ACAGUAUUUUCUAAUAACCAGUATA

[0880] Sequence ID 656-DsiRNA 324 guide (antisense) strand UAUACUGGUUAUUAGAAAAUACUGUUG

[0881] Sequence ID 657-DsiRNA 325 Passenger (Sense) Strand CAGUAUUUUCUAAUAACCAGUAUAT

[0882] Sequence ID 658-DsiRNA 325 guide (antisense) strand AUAUACUGGUUAUUAGAAAAUACUGUU

[0883] Sequence ID 659-DsiRNA 326 Passenger (Sense) Strand UUGUGAUUGUUAUCAGGAAAAAATA

[0884] Sequence ID 660-DsiRNA 326 guide (antisense) strand UAUUUUUUCCUGAUAACAAUCACAAUA

[0885] Sequence ID 661-DsiRNA 327 Passenger (Sense) Strand UGUGAUUGUUAUCAGGAAAAAAUAT

[0886] Sequence ID 662-DsiRNA 327 guide (antisense) strand AUAUUUUUUCCUGAUAACAAUCACAAU

[0887] Sequence ID 663-DsiRNA 328 Passenger (Sense) Strand GUGAUUGUUAUCAGGAAAAAAUATA

[0888] Sequence ID 664-DsiRNA 328 guide (antisense) strand UAUAUUUUUUCCUGAUAACAAUCACAA

[0889] Sequence ID 665-DsiRNA 329 Passenger (Sense) Strand UGAUUGUUAUCAGGAAAAAAUAUAT

[0890] Sequence ID 666-DsiRNA 329 guide (antisense) strand AUAUAUUUUUUCCUGAUAACAAUCACA

[0891] Sequence ID 667-DsiRNA 330 Passenger (Sense) Strand GAUUGUUAUCAGGAAAAAAUAUATT

[0892] Sequence ID 668-DsiRNA 330 guide (antisense) strand AAUAUAUUUUUUCCUGAUAACAAUCAC

[0893] Sequence ID 669-DsiRNA 331 Passenger (Sense) Strand AUUGUUAUCAGGAAAAAAUAUAUTA

[0894] Sequence ID 670-DsiRNA 331 guide (antisense) strand UAAUAUAUUUUUUCCUGAUAACAAUCA

[0895] Sequence ID 671-DsiRNA 332 Passenger (Sense) Strand UUGUUAUCAGGAAAAAAUAUAUUAA

[0896] Sequence ID 672-DsiRNA 332 guide (antisense) strand UUAAUAUAUUUUUUCCUGAUAACAAUC

[0897] Sequence ID 673-DsiRNA 333 Passenger (Sense) Strand UNOUAUCAGGAAAAAAUAUAUUAAA

[0898] Sequence ID 674-DsiRNA 333 guide (antisense) strand UUUAAUAUAUUUUUUCCUGAUAACAAU

[0899] Sequence ID 675-DsiRNA 334 Passenger (Sense) Strand GUUAUCAGGAAAAAAUAUUAAAT

[0900] Sequence ID 676-DsiRNA 334 guide (antisense) strand AUUUAAUAUAUUUUUUCCUGAUAACAA

[0901] Sequence ID 677-DsiRNA 335 Passenger (Sense) Strand UUAUCAGGAAAAAAUAUAUUAAATG

[0902] Sequence ID 678-DsiRNA 335 guide (antisense) strand CAUUUAAUAUAUUUUUUCCUGAUAACA

[0903] Sequence ID 679-DsiRNA 336 Passenger (Sense) Strand UAUCAGGAAAAAAUAUAUUAAAUGG

[0904] Sequence ID 680-DsiRNA 336 guide (antisense) strand CCAUUUAAUAUAUUUUUUCCUGAUAAC

[0905] Sequence ID 681-DsiRNA 337 Passenger (Sense) Strand AUCAGGAAAAAAUAUAUUAAAUGGC

[0906] Sequence ID 682-DsiRNA 337 guide (antisense) strand GCCAUUUAAUAUAUUUUUUCCUGAUAA

[0907] Sequence ID 683-DsiRNA 338 Passenger (Sense) Strand AGGAAAAAAUAUAUUAAAUGGCUGA

[0908] Sequence ID 684-DsiRNA 338 guide (antisense) strand UCAGCCAUUUAAUAUAUUUUUUCCUGA

[0909] Sequence ID 685-DsiRNA 339 Passenger (Sense) Strand GGAAAAAAUAUAUUAAAUGGCUGAT

[0910] Sequence ID 686-DsiRNA 339 guide (antisense) strand AUCAGCCAUUUAAUAUAUUUUUUCCUG

[0911] Sequence ID 687-DsiRNA 340 Passenger (Sense) Strand GAAAAAAUAUAUUAAAUGGCUGATA

[0912] Sequence ID 688-DsiRNA 340 guide (antisense) strand UAUCAGCCAUUUAAUAUAUUUUUUCCU

[0913] Sequence ID 689-DsiRNA 341 Passenger (Sense) Strand AAAAAAUAUAUUAAAUGGCUGAUAG

[0914] Sequence ID 690-DsiRNA 341 guide (antisense) strand CUAUCAGCCAUUUAAUAUAUUUUUUCC

[0915] Sequence ID 691-DsiRNA 342 Passenger (Sense) Strand UAUUUUCUUUCUGCUUUUAAAAATT

[0916] Sequence ID 692-DsiRNA 342 guide (antisense) strand AAUUUUUAAAAGCAGAAAGAAAAUACG

[0917] Sequence ID 693-DsiRNA 343 Passenger (Sense) Strand AUUUUCUUUCUGCUUUUAAAAAUTA

[0918] Sequence ID 694-DsiRNA 343 guide (antisense) strand UAAUUUUUAAAAGCAGAAAGAAAAUAC

[0919] Sequence ID 695-DsiRNA 344 Passenger (Sense) Strand UCUUUCUGCUUUUAAAAAUUAUUCA

[0920] Sequence ID 696-DsiRNA 344 guide (antisense) strand UGAAUAAUUUUUAAAAGCAGAAAGAAA

[0921] Sequence ID 697-DsiRNA 345 Passenger (Sense) Strand CUUUCUGCUUUUAAAAAUUAUUCAG

[0922] Sequence ID 698-DsiRNA 345 guide (antisense) strand CUGAAUAAUUUUUAAAAGCAGAAAGAA

[0923] Sequence ID 699-DsiRNA 346 Passenger (Sense) Strand UUUCUGCUUUUAAAAAUUAUUCAGG

[0924] Sequence ID 700-DsiRNA 346 guide (antisense) strand CCUGAAUAAUUUUUAAAAGCAGAAAGA

[0925] Sequence ID 701-DsiRNA 347 Passenger (Sense) Strand CUACUAAAAACACAAAAAUUAGCCA

[0926] Sequence ID 702-DsiRNA 347 guide (antisense) strand UGGCUAAUUUUUGUGUUUUUAGUAGAG

[0927] Sequence ID 703-DsiRNA 348 Passenger (Sense) Strand CAAGAUAAGGAAAUCAGGAAGUGTA

[0928] Sequence ID 704-DsiRNA 348 guide (antisense) strand UACACUUCCUGAUUUCCUUAUCUUGAU

[0929] Sequence ID 705-DsiRNA 349 Passenger (Sense) Strand AAGAUAAGGAAAUCAGGAAGUGUAA

[0930] Sequence ID 706-DsiRNA 349 guide (antisense) strand UUACACUUCCUGAUUUCCUUAUCUUGA

[0931] Sequence ID 707-DsiRNA 350 Passenger (Sense) Strand AGAUAAGGAAAUCAGGAAGUGUAAT

[0932] Sequence ID 708-DsiRNA 350 guide (antisense) strand AUUACACUUCCUGAUUUCCUUAUCUUG

[0933] Sequence ID 709-DsiRNA 351 Passenger (Sense) Strand GAUAAGGAAAUCAGGAAGUGUAATA

[0934] Sequence ID 710-DsiRNA 351 guide (antisense) strand UAUUACACUUCCUGAUUUCCUUAUCUU

[0935] Sequence ID 711-DsiRNA 352 Passenger (Sense) Strand AUAAGGAAAUCAGGAAGUGUAAUAT

[0936] Sequence ID 712-DsiRNA 352 guide (antisense) strand AUAUUACACUUCCUGAUUUCCUUAUCU

[0937] Sequence ID 713-DsiRNA 353 Passenger (Sense) Strand UAAGGAAAUCAGGAAGUGUAAUATT

[0938] Sequence ID 714-DsiRNA 353 guide (antisense) strand AAUAUUACACUUCCUGAUUUCCUUAUC

[0939] Sequence ID 715-DsiRNA 354 Passenger (Sense) Strand AAGGAAAUCAGGAAGUGUAAUAUTC

[0940] Sequence ID 716-DsiRNA 354 guide (antisense) strand GAAUAUUACACUUCCUGAUUUCCUUAU

[0941] Sequence ID 717-DsiRNA 355 Passenger (Sense) Strand AGGAAAUCAGGAAGUGUAAUAUUCT

[0942] Sequence ID 718-DsiRNA 355 guide (antisense) strand AGAAUAUUACACUUCCUGAUUUCCUUA

[0943] Sequence ID 719-DsiRNA 356 Passenger (Sense) Strand GGAAAUCAGGAAGUGUAAUAUUCTT

[0944] Sequence ID 720-DsiRNA 356 guide (antisense) strand AAGAAUAUUACACUUCCUGAUUUCCUU

[0945] Sequence ID 721-DsiRNA 357 Passenger (Sense) Strand GAAAUCAGGAAGUGUAAUAUUCUTA

[0946] Sequence ID 722-DsiRNA 357 guide (antisense) strand UAAGAAUAUUACACUUCCUGAUUUCCU

[0947] Sequence ID 723-DsiRNA 358 Passenger (Sense) Strand CUAUGAAUGCAUUCUUAUUUCUUCT

[0948] Sequence ID 724-DsiRNA 358 guide (antisense) strand AGAAGAAAUAAGAAUGCAUUCAUAGGC

[0949] Sequence ID 725-DsiRNA 359 Passenger (Sense) Strand UAUGAAUGCAUUCUUAUUUCUUCTT

[0950] Sequence ID 726-DsiRNA 359 guide (antisense) strand AAGAAGAAAUAAGAAUGCAUUCAUAGG

[0951] Sequence ID 727-DsiRNA 360 Passenger (Sense) Strand CUACCACACCCAGCUAGUUUUUUTT

[0952] Sequence ID 728-DsiRNA 360 guide (antisense) strand AAAAAAAACUAGCUGGGUGUGGUAGUG

[0953] Sequence ID 729-DsiRNA 361 Passenger (Sense) Strand CCACACCCAGCUAGUUUUUUUUUGT

[0954] Sequence ID 730-DsiRNA 361 guide (antisense) strand ACAAAAAAAAACUAGCUGGGUGUGGUA

[0955] Sequence ID 731-DsiRNA 362 Passenger (Sense) Strand CACACCCAGCUAGUUUUUUUUUGTA

[0956] Sequence ID 732-DsiRNA 362 guide (antisense) strand UACAAAAAAAAACUAGCUGGGUGUGGU

[0957] Sequence ID 733-DsiRNA 363 Passenger (Sense) Strand GCUAGGAUUACAGGUGUGAGCUACC

[0958] Sequence ID 734-DsiRNA 363 guide (antisense) strand GGUAGCUCACACCUGUAAUCCUAGCAC

[0959] Sequence ID 735-DsiRNA 364 Passenger (Sense) Strand CUAGGAUUACAGGUGUGAGCUACCA

[0960] Sequence ID 736-DsiRNA 364 guide (antisense) strand UGGUAGCUCACACCUGUAAUCCUAGCA

[0961] Sequence ID 737-DsiRNA 365 Passenger (Sense) Strand CCAUGCCUGGUCCAACAUUCUUCAT

[0962] Sequence ID 738-DsiRNA 365 guide (antisense) strand AUGAAGAAUGUUGGACCAGGCAUGGUA

[0963] Sequence ID 739-DsiRNA 366 passenger (sense) strand UGCAGAGUAUGAGCCUGAUUUUGTT

[0964] Sequence ID 740-DsiRNA 366 guide (antisense) strand AACAAAAUCAGGCUCAUACUCUGCACU

[0965] Sequence ID 741-DsiRNA 367 Passenger (Sense) Strand GCAGAGUAUGAGCCUGAUUUUGUTT

[0966] Sequence ID 742-DsiRNA 367 guide (antisense) strand AAACAAAAUCAGGCUCAUACUCUGCAC

[0967] Sequence ID 743-DsiRNA 368 Passenger (Sense) Strand CAGAGUAUGAGCCUGAUUUUGUUTA

[0968] Sequence ID 744-DsiRNA 368 guide (antisense) strand UAAACAAAAUCAGGCUCAUACUCUGCA

[0969] Sequence ID 745-DsiRNA 369 Passenger (Sense) Strand AGAGUAUGAGCCUGAUUUUGUUUAA

[0970] Sequence ID 746-DsiRNA 369 guide (antisense) strand UUAAACAAAAUCAGGCUCAUACUCUGC

[0971] Sequence ID 747-DsiRNA 370 Passenger (Sense) Strand GAGUAUGAGCCUGAUUUUGUUUAAA

[0972] Sequence ID 748-DsiRNA 370 guide (antisense) strand UUUAAACAAAAUCAGGCUCAUACUCUG

[0973] Sequence ID 749-DsiRNA 371 Passenger (Sense) GGGUGAAACCCCAUCUCUACUAAAA

[0974] Sequence ID 750-DsiRNA 371 guide (antisense) strand UUUUAGUAGAGAUGGGGUUUCACCCAG

[0975] Sequence ID 751-DsiRNA 372 Passenger (Sense) Strand GUGAAACCCCAUCUCUACUAAAAAA

[0976] Sequence ID 752-DsiRNA 372 guide (antisense) strand UUUUUUAGUAGAGAUGGGGUUUCACCC

[0977] Sequence ID 753-DsiRNA 373 Passenger (Sense) Strand UGAAACCCCAUCUCUACUAAAAAAT

[0978] Sequence ID 754-DsiRNA 373 guide (antisense) strand AUUUUUUAGUAGAGAUGGGGUUUCACC

[0979] Sequence ID 755-DsiRNA 374 Passenger (Sense) Strand GAAACCCCAUCUCUACUAAAAAATG

[0980] Sequence ID 756-DsiRNA 374 guide (antisense) strand CAUUUUUUAGUAGAGAUGGGGUUUCAC

[0981] Sequence ID 757-DsiRNA 375 guide (antisense) strand AAACCCCAUCUCUACUAAAAAAUGC

[0982] Sequence ID 758-DsiRNA 375 guide (antisense) strand GCAUUUUUUAGUAGAGAUGGGGUUUCA

[0983] Sequence ID 759-DsiRNA 376 Passenger (Sense) Strand AACCCCAUCUCUACUAAAAAAUGCA

[0984] Sequence ID 760-DsiRNA 376 guide (antisense) strand UGCAUUUUUUAGUAGAGAUGGGGUUUC

[0985] Sequence ID 761-DsiRNA 377 Passenger (Sense) Strand CCCAUCUCUACUAAAAAAUGCAAAA

[0986] Sequence ID 762-DsiRNA 377 guide (antisense) strand UUUUGCAUUUUUUAGUAGAGAUGGGGU

[0987] Sequence ID 763-DsiRNA 378 Passenger (Sense) Strand AUCAAAACCCUUAUGGCAGACUGTT

[0988] Sequence ID 764-DsiRNA 378 guide (antisense) strand AACAGUCUGCCAUAAGGGUUUUGAUAU

[0989] Sequence ID 765-DsiRNA 379 Passenger (Sense) Strand UAUUUUAUUUGUCGUGCUUAUAUGT

[0990] Sequence ID 766-DsiRNA 379 guide (antisense) strand ACAUAUAAGCACGACAAAUAAAAUACA

[0991] Sequence ID 767-DsiRNA 380 Passenger (Sense) Strand GUGUUGCCCAAGUUUCUAUGGUGAA

[0992] Sequence ID 768-DsiRNA 380 guide (antisense) strand UUCACCAUAGAAACUUGGGCAACACAU

[0993] Sequence ID 769-DsiRNA 381 Passenger (Sense) Strand GCCCAAGUUUCUAUGGUGAACGGTA

[0994] Sequence ID 770-DsiRNA 381 guide (antisense) strand UACCGUUCACCAUAGAAACUUGGGCAA

[0995] Sequence ID 771-DsiRNA 382 Passenger (Sense) Strand CCCAAGUUUCUAUGGUGAACGGUAT

[0996] Sequence ID 772-DsiRNA 382 guide (antisense) strand AUACCGUUCACCAUAGAAACUUGGGCA

[0997] Sequence ID 773-DsiRNA 383 Passenger (Sense) Strand ACUUUCAGCAUGAGAAAAUAACUCC

[0998] Sequence ID 774-DsiRNA 383 guide (antisense) strand GGAGUUAUUUUCUCAUGCUGAAAGUGA

[0999] Sequence ID 775-DsiRNA 384 Passenger (Sense) Strand CUUUCAGCAUGAGAAAAUAACUCCT

[1000] Sequence ID 776-DsiRNA 384 guide (antisense) strand AGGAGUUAUUUUCUCAUGCUGAAAGUG

[1001] Sequence ID 777-PNPLA3 oligonucleotide 1 passenger (sense) chain UGAGUGACAACGUACCCUUAGCAGCCCGAAAGGCUGC

[1002] Sequence ID 778-PNPLA3 oligonucleotide-1 guide (antisense) chain UAAGGGUACGUUGUCACUCAGG

[1003] Sequence ID 779-PNPLA3 oligonucleotide 2 passenger (sense) chain GAGUGACAACGUACCCUUCAGCAGCCGAAAGGCUGC

[1004] Sequence ID 780-PNPLA3 oligonucleotide 2 guide (antisense) chain UGAAGGGUACGUUGUCACUCGG

[1005] Sequence ID 781-PNPLA3 oligonucleotide 3 passenger (sense) chain AGUGACAACGUACCCUUCAAGCAGCCGAAAGGCUGC

[1006] Sequence ID 782-PNPLA3 oligonucleotide 3 guide (antisense) chain UUGAAGGGUACGUUGUCACUGG

[1007] Sequence ID 783-PNPLA3 oligonucleotide 4 passenger (sense) chain UGACAACGUACCCUUCAUUAGCAGCCCGAAAGGCUGC

[1008] Sequence ID 784-PNPLA3 oligonucleotide 4 guide (antisense) chain UAAUGAAGGGUACGUUGUCAGG

[1009] Sequence ID 785-PNPLA3 oligonucleotide 5 passenger (sense) chain GACAACGUACCCUUCAUUGAGCAGCCGAAAGGCUGC

[1010] Sequence ID 786-PNPLA3 oligonucleotide 5 guide (antisense) chain UCAAUGAAGGGUACGUUGUCGG

[1011] Sequence ID 787-PNPLA3 oligonucleotide 6 passenger (sense) chain CAACGUACCCUUCAUUGAUAGCAGCCGAAAGGCUGC

[1012] Sequence ID 788-PNPLA3 oligonucleotide 6 guide (antisense) chain UAUCAAUGAAGGGUACGUUGGG

[1013] Sequence ID 789-PNPLA3 oligonucleotide 7 passenger (sense) chain AACGUACCCUUCAUUGAUGAGCAGCCGAAAGGCUGC

[1014] Sequence ID 790-PNPLA3 oligonucleotide 7 guide (antisense) chain UCAUCAAUGAAGGGUACGUUGG

[1015] Sequence ID 791-PNPLA3 oligonucleotide 8 passenger (sense) chain ACGUACCCUUCAUUGAUGCAGCAGCCGAAAGGCUGC

[1016] Sequence ID 792-PNPLA3 oligonucleotide 8 guide (antisense) chain UGCAUCAAUGAAGGGUACGUGG

[1017] Sequence ID 793-PNPLA3 oligonucleotide 9 passenger (sense) chain CGUACCCUUCAUUGAUGCCAGCAGCCCGAAAGGCUGC

[1018] Sequence ID 794-PNPLA3 oligonucleotide 9 guide (antisense) chain UGGCAUCAAUGAAGGGUACGGG

[1019] Sequence ID 795-PNPLA3 oligonucleotide 10 passenger (sense) chain UACCCUUCAUUGAUGCCAAAGCAGCCGAAAGGCUGC

[1020] Sequence ID 796-PNPLA3 oligonucleotide 10 guide (antisense) chain UUUGGCAUCAAUGAAGGGUAGG

[1021] Sequence ID 797-PNPLA3 oligonucleotide-11 passenger (sense) chain CACGAACUUUCUUCAUGUGAGCAGCCGAAAGGCUGC

[1022] Sequence ID 798-PNPLA3 oligonucleotide-11 guide (antisense) chain UCACAUGAAGAAAGUUCGUGGG

[1023] Sequence ID 799-PNPLA3 oligonucleotide-12 passenger (sense) chain ACGAACUUUCUUCAUGUGGAGCAGCCGAAAGGCUGC

[1024] Sequence ID 800-PNPLA3 oligonucleotide-12 guide (antisense) chain UCCACAUGAAGAAAGUUCGUGG

[1025] Sequence ID 801-PNPLA3 oligonucleotide 13 passenger (sense) chain CGAACUUUCUUCAUUGUGGAAGCAGCCGAAAGGCUGC

[1026] Sequence ID 802-PNPLA3 oligonucleotide-13 guide (antisense) chain UUCCACAUGAAGAAAGUUCGGG

[1027] Sequence ID 803-PNPLA3 oligonucleotide 14 passenger (sense) chain ACUUUCUUCAUGUGGACAUAAGCAGCCGAAAGGCUGC

[1028] Sequence ID 804-PNPLA3 oligonucleotide 14 guide (antisense) chain UAUGUCCACAUGAAGAAAGUGG

[1029] Sequence ID 805-PNPLA3 oligonucleotide 15 passenger (sense) chain UUUCUUCAUGUGGACAUCAAGCAGCCGAAAGGCUGC

[1030] Sequence ID 806-PNPLA3 oligonucleotide 15 guide (antisense) chain UUGAUGUCCACAUGAAGAAAGG

[1031] Sequence ID 807-PNPLA3 oligonucleotide 16 passenger (sense) chain CUUCAUGUGGACAUCACCAAGCAGCCGAAAGGCUGC

[1032] Sequence ID 808-PNPLA3 oligonucleotide 16 guide (antisense) chain UUGGUGAUGUCCACAUGAAGGG

[1033] Sequence ID 809-PNPLA3 oligonucleotide 17 passenger (sense) chain AUGUGGACAUCACCAAGCUAGCAGCCGAAAGGCUGC

[1034] Sequence ID 810-PNPLA3 oligonucleotide 17 guide (antisense) chain UAGCUUGGUGAUGUCCACAUGG

[1035] Sequence ID 811-PNPLA3 oligonucleotide 18 passenger (sense) chain UGUGGACAUCACCAAGCUCAGCAGCCGAAAGGCUGC

[1036] Sequence ID 812-PNPLA3 oligonucleotide-18 guide (antisense) chain UGAGCUUGGUGAUGUCCACAGG

[1037] Sequence ID 813-PNPLA3 oligonucleotide-19 passenger (sense) chain GUGGACAUCACCAAGCUCAAGCAGCCGAAAGGCUGC

[1038] Sequence ID 814-PNPLA3 oligonucleotide-19 guide (antisense) chain UUGAGCUUGGUGAUGUCCACGG

[1039] Sequence ID 815-PNPLA3 oligonucleotide 20 passenger (sense) chain UGGACAUCACCAAGCUCAGAGCAGCCGAAAGGCUGC

[1040] Sequence ID 816-PNPLA3 oligonucleotide 20 guide (antisense) chain UCUGAGCUUGGUGAUGUCCAGG

[1041] Sequence ID 817-PNPLA3 oligonucleotide-21 passenger (sense) chain AGAUAUGCCUUCGAGGAUAAGCAGCCCGAAAGGCUGC

[1042] Sequence ID 818-PNPLA3 oligonucleotide-21 guide (antisense) chain UUAUCCUCGAAGGCAUAUCUGG

[1043] Sequence ID 819-PNPLA3 oligonucleotide-22 passenger (sense) chain AUGCCUUCGAGGAUAUUUGAGCAGCCGAAAGGCUGC

[1044] Sequence ID 820-PNPLA3 oligonucleotide-22 guide (antisense) chain UCAAAUAUCCUCGAAGGCAUGG

[1045] Sequence ID 821-PNPLA3 oligonucleotide 23 passenger (sense) chain GCCUUCGAGGAUAUUUGGAAGCAGCCGAAAGGCUGC

[1046] Sequence ID 822-PNPLA3 oligonucleotide-23 guide (antisense) chain UUCCAAAUAUCCUCGAAGGCGG

[1047] Sequence ID 823-PNPLA3 oligonucleotide 24 passenger (sense) chain UGAAGUCAUCCUCAGAAGGAGCAGCCGAAAGGCUGC

[1048] Sequence ID 824-PNPLA3 oligonucleotide 24 guide (antisense) chain UCCUUCUGAGGAUGACUUCAGG

[1049] Sequence ID 825-PNPLA3 oligonucleotide 25 passenger (sense) chain GAAGUCAUCCUCAGAAGGGAGCAGCCGAAAGGCUGC

[1050] Sequence ID 826-PNPLA3 oligonucleotide-25 guide (antisense) chain UCCCUUCUGAGGAUGACUUCGG

[1051] Sequence ID 827-PNPLA3 oligonucleotide 26 passenger (sense) chain AUCCUCAGAAGGGAUGGAUAGCAGCCGAAAGGCUGC

[1052] Sequence ID 828-PNPLA3 oligonucleotide 26 guide (antisense) chain UAUCCAUCCCUUCUGAGGAUGG

[1053] Sequence ID 829-PNPLA3 oligonucleotide 27 passenger (sense) chain AGUGAAGAAAUGAAAGACAAGCAGCCGAAAGGCUGC

[1054] Sequence ID 830-PNPLA3 oligonucleotide 27 guide (antisense) chain UUGUCUUUCAUUUCUUCACUGG

[1055] Sequence ID 831-PNPLA3 oligonucleotide 28 passenger (sense) chain AAGAAAUGAAAGACAAAGGAGCAGCCGAAAGGCUGC

[1056] Sequence ID 832-PNPLA3 oligonucleotide 28 guide (antisense) chain UCCUUUGUCUUUCAUUUCUUGG

[1057] Sequence ID 833-PNPLA3 oligonucleotide 29 passenger (sense) chain AGAAAUGAAAGACAAAGGUAGCAGCCGAAAGGCUGC

[1058] Sequence ID 834-PNPLA3 oligonucleotide 29 guide (antisense) chain UACCUUUGUCUUUCAUUUCUGG

[1059] Sequence ID 835-PNPLA3 oligonucleotide 30 passenger (sense) chain GAAAUGAAAGACAAAGGUGAGCAGCCGAAAGGCUGC

[1060] Sequence ID 836-PNPLA3 oligonucleotide 30 guide (antisense) chain UCACCUUUGUCUUUCAUUUCGG

[1061] Sequence ID 837-PNPLA3 oligonucleotide 31 passenger (sense) chain AAAUGAAAGACAAAGGUGGAGCAGCCGAAAGGCUGC

[1062] Sequence ID 838-PNPLA3 oligonucleotide 31 guide (antisense) chain UCCACCUUUGUCUUUCAUUUGG

[1063] Sequence ID 839-PNPLA3 oligonucleotide 32 passenger (sense) chain AAUGAAAGACAAAGGUGGAAGCAGCCGAAAGGCUGC

[1064] Sequence ID 840-PNPLA3 oligonucleotide 32 guide (antisense) chain UUCCACCUUUGUCUUUCAUUGG

[1065] Sequence ID 841-PNPLA3 oligonucleotide 33 passenger (sense) chain AUGAAAGACAAAGGUGGAUAGCAGCCGAAAGGCUGC

[1066] Sequence ID 842-PNPLA3 oligonucleotide 33 guide (antisense) chain UAUCCACCUUUGUCUUUCAUGG

[1067] Sequence ID 843-PNPLA3 oligonucleotide 34 passenger (sense) chain UGAAAGACAAAGGUGGAUAAGCAGCCGAAAGGCUGC

[1068] Sequence ID 844-PNPLA3 oligonucleotide 34 guide (antisense) chain UUAUCCACCUUUGUCUUUCAGG

[1069] Sequence ID 845-PNPLA3 oligonucleotide 35 passenger (sense) chain GAAAGACAAAGGUGGAUACAGCAGCCGAAAGGCUGC

[1070] Sequence ID 846-PNPLA3 oligonucleotide 35 guide (antisense) chain UGUAUCCACCUUUGUCUUUCGG

[1071] Sequence ID 847-PNPLA3 oligonucleotide 36 passenger (sense) chain AAGACAAAGGUGGAUACAUAGCAGCCCGAAAGGCUGC

[1072] Sequence ID 848-PNPLA3 oligonucleotide 36 guide (antisense) chain UAUGUAUCCACCUUUGUCUUGG

[1073] Sequence ID 849-PNPLA3 oligonucleotide 37 passenger (sense) chain AGACAAAGGUGGAUACAUGAGCAGCCGAAAGGCUGC

[1074] Sequence ID 850-PNPLA3 oligonucleotide 37 guide (antisense) chain UCAUGUAUCCACCUUUGUCUGG

[1075] Sequence ID 851-PNPLA3 oligonucleotide 38 passenger (sense) chain GACAAAGGUGGAUACAUGAAGCAGCCCGAAAGGCUGC

[1076] Sequence ID 852-PNPLA3 oligonucleotide 38 guide (antisense) chain UUCAUGUAUCCACCUUUGUCGG

[1077] Sequence ID 853-PNPLA3 oligonucleotide 39 passenger guide (sense) chain ACAAAGGUGGAUACAUGAGAGCAGCCGAAAGGCUGC

[1078] Sequence ID 854-PNPLA3 oligonucleotide 39 guide (antisense) chain UCUCAUGUAUCCACCUUUGUGG

[1079] Sequence ID 855-PNPLA3 oligonucleotide 40 passenger (sense) chain AAGGUGGAUACAUGAGCAAAGCAGCCGAAAGGCUGC

[1080] Sequence ID 856-PNPLA3 oligonucleotide 40 guide (antisense) chain UUUGCUCAUGUAUCCACCUUGG

[1081] Sequence ID 857-PNPLA3 oligonucleotide 41 passenger (sense) chain AGGUGGAUACAUGAGCAAGAGCAGCCGAAAGGCUGC

[1082] Sequence ID 858-PNPLA3 oligonucleotide 41 guide (antisense) chain UCUUGCUCAUGUAUCCACCUGG

[1083] Sequence ID 859-PNPLA3 oligonucleotide 42 passenger (sense) chain UGGAUACAUGAGCAAGAUUAGCAGCCCGAAAGGCUGC

[1084] Sequence ID 860-PNPLA3 oligonucleotide 42 guide (antisense) chain UAAUCUUGCUCAUGUAUCCAGG

[1085] Sequence ID 861-PNPLA3 oligonucleotide 43 passenger (sense) chain GGAUACAUGAGCAAGAUUUAGCAGCCCGAAAGGCUGC

[1086] Sequence ID 862-PNPLA3 oligonucleotide 43 guide (antisense) chain

[1087] UAAAUCUUGCUCAUGUAUCCGG

[1088] Sequence ID 863-PNPLA3 oligonucleotide 44 passenger (sense) chain GAUACAUGAGCAAGAUUUGAGCAGCCGAAAGGCUGC

[1089] Sequence ID 864-PNPLA3 oligonucleotide 44 guide (antisense) chain UCAAAUCUUGCUCAUGUAUCGG

[1090] Sequence ID 865-PNPLA3 oligonucleotide 45 passenger (sense) chain AUACAUGAGCAAGAUUUGCAGCAGCCGAAAGGCUGC

[1091] Sequence ID 866-PNPLA3 oligonucleotide 45 guide (antisense) chain UGCAAAUCUUGCUCAUGUAUGG

[1092] Sequence ID 867-PNPLA3 oligonucleotide 46 passenger (sense) chain UACAUGAGCAAGAUUUGCAAGCAGCCGAAAGGCUGC

[1093] Sequence ID 868-PNPLA3 oligonucleotide 46 guide (antisense) chain UUGCAAAUCUUGCUCAUGUAGG

[1094] Sequence ID 869-PNPLA3 oligonucleotide 47 passenger (sense) chain ACAUGAGCAAGAUUUGCAAAGCAGCCGAAAGGCUGC

[1095] Sequence ID 870-PNPLA3 oligonucleotide 47 guide (antisense) chain UUUGCAAAUCUUGCUCAUGUGG

[1096] Sequence ID 871-PNPLA3 oligonucleotide 48 passenger (sense) chain UGAGCAAGAUUUGCAACUUAGCAGCCCGAAAGGCUGC

[1097] Sequence ID 872-PNPLA3 oligonucleotide 48 guide (antisense) chain UAAGUUGCAAAUCUUGCUCAGG

[1098] Sequence ID 873-PNPLA3 oligonucleotide 49 passenger (sense) chain GAGCAAGAUUUGCAACUUGAGCAGCCGAAAGGCUGC

[1099] Sequence ID 874-PNPLA3 oligonucleotide 49 guide (antisense) chain UCAAGUUGCAAAUCUUGCUCGG

[1100] Sequence ID 875-PNPLA3 oligonucleotide 50 passenger (sense) chain AGCAAGAUUUGCAACUUGCAGCAGCCGAAAGGCUGC

[1101] Sequence ID 876-PNPLA3 oligonucleotide 50 guide (antisense) chain UGCAAGUUGCAAAUCUUGCUGG

[1102] Sequence ID 877-PNPLA3 oligonucleotide 51 passenger (sense) chain GCAAGAUUUGCAACUUGCUAGCAGCCGAAAGGCUGC

[1103] Sequence ID 878-PNPLA3 oligonucleotide 51 guide (antisense) chain UAGCAAGUUGCAAAUCUUGCGG

[1104] Sequence ID 879-PNPLA3 oligonucleotide 52 passenger (sense) chain CAAGAUUUGCAACUUGCUAAGCAGCCCGAAAGGCUGC

[1105] Sequence ID 880-PNPLA3 oligonucleotide 52 guide (antisense) chain UUAGCAAGUUGCAAAUCUUGGG

[1106] Sequence ID 881-PNPLA3 oligonucleotide 53 passenger (sense) chain AAGAUUUGCAACUUGCUACAGCAGCCGAAAGGCUGC

[1107] Sequence ID 882-PNPLA3 oligonucleotide 53 guide (antisense) chain UGUAGCAAGUUGCAAAUCUUGG

[1108] Sequence ID 883-PNPLA3 oligonucleotide 54 passenger (sense) chain AGAUUUGCAACUUGCUACCAGCAGCCGAAAGGCUGC

[1109] Sequence ID 884-PNPLA3 oligonucleotide 54 guide (antisense) chain UGGUAGCAAGUUGCAAAUCUGG

[1110] Sequence ID 885-PNPLA3 oligonucleotide 55 passenger (sense) chain AUUUGCAACUUGCUACCCAAGCAGCCGAAAGGCUGC

[1111] Sequence ID 886-PNPLA3 oligonucleotide 55 guide (antisense) chain UUGGGUAGCAAGUUGCAAAUGG

[1112] Sequence ID 887-PNPLA3 oligonucleotide 56 passenger (sense) chain UUGCAACUUGCUACCCAUUAGCAGCCCGAAAGGCUGC

[1113] Sequence ID 888-PNPLA3 oligonucleotide 56 guide (antisense) chain UAAUGGGUAGCAAGUUGCAAGG

[1114] Sequence ID 889-PNPLA3 oligonucleotide 57 passenger (sense) chain UGCAACUUGCUACCCAUUAAGCAGCCGAAAGGCUGC

[1115] Sequence ID 890-PNPLA3 oligonucleotide 57 guide (antisense) chain UUAAUGGGUAGCAAGUUGCAGG

[1116] Sequence ID 891-PNPLA3 oligonucleotide 58 passenger (sense) chain GCAACUUGCUACCCAUUAGAGCAGCCCGAAAGGCUGC

[1117] Sequence ID 892-PNPLA3 oligonucleotide 58 guide (antisense) chain UCUAAUGGGUAGCAAGUUGCGG

[1118] Sequence ID 893-PNPLA3 oligonucleotide 59 passenger (sense) chain CAACUUGCUACCCAUUAGGAGCAGCCGAAAGGCUGC

[1119] Sequence ID 894-PNPLA3 oligonucleotide 59 guide (antisense) chain UCCUAAUGGGUAGCAAGUUGGG

[1120] Sequence ID 895-PNPLA3 oligonucleotide 60 passenger (sense) chain CUUGCUACCCAUUAGGAUAAGCAGCCCGAAAGGCUGC

[1121] Sequence ID 896-PNPLA3 oligonucleotide 60 guide (antisense) chain UUAUCCUAAUGGGUAGCAAGGG

[1122] Sequence ID 897-PNPLA3 oligonucleotide 61 passenger (sense) chain UGCUACCCAUUAGGAUAAUAGCAGCCGAAAGGCUGC

[1123] Sequence ID 898-PNPLA3 oligonucleotide 61 guide (antisense) chain UAUUAUCCUAAUGGGUAGCAGG

[1124] Sequence ID 899-PNPLA3 oligonucleotide 62 passenger (sense) chain AUUAGGAUAAUGUCUUAUGAGCAGCCGAAAGGCUGC

[1125] Sequence ID 900-PNPLA3 oligonucleotide 62 guide (antisense) chain UCAUAAGACAUUAUCCUAAUGG

[1126] Sequence ID 901-PNPLA3 oligonucleotide 63 passenger (sense) chain UUAGGAUAAUGUCUUAUGUAGCAGCCGAAAGGCUGC

[1127] Sequence ID 902-PNPLA3 oligonucleotide 63 guide (antisense) chain UACAUAAGACAUUAUCCUAAGG

[1128] Sequence ID 903-PNPLA3 oligonucleotide 64 passenger (sense) chain UGCGAUUGUCCAGAGACUGAGCAGCCGAAAGGCUGC

[1129] Sequence ID 904-PNPLA3 oligonucleotide 64 guide (antisense) chain UCAGUCUCUGGACAAUCGCAGG

[1130] Sequence ID 905-PNPLA3 oligonucleotide 65 passenger (sense) chain CGAUUGUCCAGAGACUGGUAGCAGCCGAAAGGCUGC

[1131] Sequence ID 906-PNPLA3 oligonucleotide 65 guide (antisense) chain UACCAGUCUCUGGACAAUCGGG

[1132] Sequence ID 907-PNPLA3 oligonucleotide 66 passenger (sense) chain GUCCAGAGACUGGUGACAUAAGCAGCCGAAAGGCUGC

[1133] Sequence ID 908-PNPLA3 oligonucleotide 66 guide (antisense) chain UAUGUCACCAGUCUCUGGACGG

[1134] Sequence ID 909-PNPLA3 oligonucleotide 67 passenger (sense) chain AGAGACUGGUGACAUGGCUAGCAGCCGAAAGGCUGC

[1135] Sequence ID 910-PNPLA3 oligonucleotide 67 guide (antisense) chain UAGCCAUGUCACCAGUCUCUGG

[1136] Sequence ID 911-PNPLA3 oligonucleotide 68 passenger (sense) chain ACUGGUGACAUGGCUUCCAAGCAGCCGAAAGGCUGC

[1137] Sequence ID 912-PNPLA3 oligonucleotide 68 guide (antisense) chain UUGGAAGCCAUGUCACCAGUGG

[1138] Sequence ID 913-PNPLA3 oligonucleotide 69 passenger (sense) chain CUCCACCUUUCCCAGUUUUAGCAGCCGAAAGGCUGC

[1139] Sequence ID 914-PNPLA3 oligonucleotide 69 guide (antisense) chain UAAAACUGGGAAAGGUGGAGGG

[1140] Sequence ID 915-PNPLA3 oligonucleotide 70 passenger (sense) chain CACCUUUCCCAGUUUUUCAAGCAGCCGAAAGGCUGC

[1141] Sequence ID 916-PNPLA3 oligonucleotide 70 guide (antisense) chain UUGAAAAACUGGGAAAGGUGGG

[1142] Sequence ID 917-PNPLA3 oligonucleotide 71 passenger (sense) chain AGUUUUUCACUAGAGAAGAAGCAGCCGAAAGGCUGC

[1143] Sequence ID 918-PNPLA3 oligonucleotide 71 guide (antisense) chain UUCUUCUCUAGUGAAAAACUGG

[1144] Sequence ID 919-PNPLA3 oligonucleotide 72 passenger (sense) chain AGCAGAUUCUUUCAGAGGUAGCAGCCGAAAGGCUGC

[1145] Sequence ID 920-PNPLA3 oligonucleotide 72 guide (antisense) chain UACCUCUGAAAGAAUCUGCUGG

[1146] Sequence ID 921-PNPLA3 oligonucleotide 73 passenger (sense) chain GCAGAUUCUUUCAGAGGUGAGCAGCCGAAAGGCUGC

[1147] Sequence ID 922-PNPLA3 oligonucleotide 73 guide (antisense) chain UCACCUCUGAAAGAAUCUGCGG

[1148] Sequence ID 923-PNPLA3 oligonucleotide 74 passenger (sense) chain AGAUUCUUUCAGAGGUGCUAGCAGCCGAAAGGCUGC

[1149] Sequence ID 924-PNPLA3 oligonucleotide 74 guide (antisense) chain UAGCACCUCUGAAAGAAUCUGG

[1150] Sequence ID 925-PNPLA3 oligonucleotide 75 passenger (sense) chain AGGUGCUAAAGUUUCCCAUAGCAGCCGAAAGGCUGC

[1151] Sequence ID 926-PNPLA3 oligonucleotide 75 guide (antisense) chain UAUGGGAAACUUUAGCACCUGG

[1152] Sequence ID 927-PNPLA3 oligonucleotide 76 passenger (sense) chain AAAGUUUCCCAUCUUUGUGAGCAGCCGAAAGGCUGC

[1153] Sequence ID 928-PNPLA3 oligonucleotide 76 guide (antisense) chain UCACAAAGAUGGGAAACUUUGG

[1154] Sequence ID 929-PNPLA3 oligonucleotide 77 passenger (sense) chain AGUUUCCCAUCUUUGUGCAAGCAGCCGAAAGGCUGC

[1155] Sequence ID 930-PNPLA3 oligonucleotide 77 guide (antisense) chain UUGCACAAAGAUGGGAAACUGG

[1156] Sequence ID 931-PNPLA3 oligonucleotide 78 passenger (sense) chain AGCCUCUGAGCUGAGUUGGAGCAGCCGAAAGGCUGC

[1157] Sequence ID 932-PNPLA3 oligonucleotide 78 guide (antisense) chain UCCAACUCAGCUCAGAGGCUGG

[1158] Sequence ID 933-PNPLA3 oligonucleotide 79 passenger (sense) chain GCCUCUGAGCUGAGUUGGUAGCAGCCGAAAGGCUGC

[1159] Sequence ID 934-PNPLA3 oligonucleotide 79 guide (antisense) chain UACCAACUCAGCUCUCAGAGGCGG

[1160] Sequence ID 935-PNPLA3 oligonucleotide 80 passenger (sense) chain CCUCUGAGCUGAGUUGGUUAGCAGCCCGAAAGGCUGC

[1161] Sequence ID 936-PNPLA3 oligonucleotide 80 guide (antisense) chain UAACCAACUCAGCUCAGAGGGG

[1162] Sequence ID 937-PNPLA3 oligonucleotide 81 passenger (sense) chain CUCUGAGCUGAGUUGGUUUAGCAGCCCGAAAGGCUGC

[1163] Sequence ID 938-PNPLA3 oligonucleotide 81 guide (antisense) chain UAAACCAACUCAGCUCAGAGGG

[1164] Sequence ID 939-PNPLA3 oligonucleotide 82 passenger (sense) chain UCUGAGCUGAGUUGGUUUUAGCAGCCCGAAAGGCUGC

[1165] Sequence ID 940-PNPLA3 oligonucleotide-82 guide (antisense) chain UAAAACCAACUCAGCUCAGAGG

[1166] Sequence ID 941-PNPLA3 oligonucleotide 83 passenger (sense) chain UGAGCUGAGUUGGUUUUAUAGCAGCCGAAAGGCUGC

[1167] Sequence ID 942-PNPLA3 oligonucleotide-83 guide (antisense) chain UAUAAAACCAACUCAGCUCAGG

[1168] Sequence ID 943-PNPLA3 oligonucleotide 84 passenger (sense) chain UGAGUUGGUUUUAUGAAAAAGCAGCCGAAAGGCUGC

[1169] Sequence ID 944-PNPLA3 oligonucleotide 84 guide (antisense) chain UUUUUCAUAAAACCAACUCAGG

[1170] Sequence ID 945-PNPLA3 oligonucleotide 85 passenger (sense) chain GAGUUGGUUUUAUGAAAAGAGCAGCCGAAAGGCUGC

[1171] Sequence ID 946-PNPLA3 oligonucleotide 85 guide (antisense) chain UCUUUUCAUAAAACCAACUCGG

[1172] Sequence ID 947-PNPLA3 oligonucleotide 86 passenger (sense) chain UUGGUUUUAUGAAAAGCUAAGCAGCCCGAAAGGCUGC

[1173] Sequence ID 948-PNPLA3 oligonucleotide 86 guide (antisense) chain UUAGCUUUUCAUAAAACCAAGG

[1174] Sequence ID 949-PNPLA3 oligonucleotide 87 passenger (sense) chain UGGUUUUAUGAAAAGCUAGAGCAGCCGAAAGGCUGC

[1175] Sequence ID 950-PNPLA3 oligonucleotide 87 guide (antisense) chain UCUAGCUUUUCAUAAAACCAGG

[1176] Sequence ID 951-PNPLA3 oligonucleotide 88 passenger (sense) chain GGUUUUAUGAAAAGCUAGGAGCAGCCGAAAGGCUGC

[1177] Sequence ID 952-PNPLA3 oligonucleotide 88 guide (antisense) chain UCCUAGCUUUUCAUAAAACCGG

[1178] Sequence ID 953-PNPLA3 oligonucleotide 89 passenger (sense) chain UUUUAUGAAAAGCUAGGAAAGCAGCCGAAAGGCUGC

[1179] Sequence ID 954-PNPLA3 oligonucleotide 89 guide (antisense) chain UUUCCUAGCUUUUCAUAAAAGG

[1180] Sequence ID 955-PNPLA3 oligonucleotide 90 passenger (sense) chain UAUGAAAAGCUAGGAAGCAAGCAGCCGAAAGGCUGC

[1181] Sequence ID 956-PNPLA3 oligonucleotide 90 guide (antisense) chain UUGCUUCCUAGCUUUUCAUAGG

[1182] Sequence ID 957-PNPLA3 oligonucleotide 91 passenger (sense) chain AUGAAAAGCUAGGAAGCAAAGCAGCCGAAAGGCUGC

[1183] Sequence ID 958-PNPLA3 oligonucleotide 91 guide (antisense) chain UUUGCUUCCUAGCUUUUCAUGG

[1184] Sequence ID 959-PNPLA3 oligonucleotide 92 passenger (sense) chain CCAGCACUUAACUCUAAUAAGCAGCCGAAAGGCUGC

[1185] Sequence ID 960-PNPLA3 oligonucleotide 92 guide (antisense) chain UUAUUAGAGUUAAGUGCUGGGG

[1186] Sequence ID 961-PNPLA3 oligonucleotide 93 passenger (sense) chain CAGCACUUAACUCUAAUACAGCAGCCGAAAGGCUGC

[1187] Sequence ID 962-PNPLA3 oligonucleotide 93 guide (antisense) chain UGUAUUAGAGUUAAGUGCUGGG

[1188] Sequence ID 963-PNPLA3 oligonucleotide 94 passenger (sense) chain UAAUACAUCAGCAUGCGUUAGCAGCCCGAAAGGCUGC

[1189] Sequence ID 964-PNPLA3 oligonucleotide 94 guide (antisense) chain UAACGCAUGCUGAUGUAUUAGG

[1190] Sequence ID 965-PNPLA3 oligonucleotide 95 passenger (sense) chain AAUACAUCAGCAUGCGUUAAGCAGCCGAAAGGCUGC

[1191] Sequence ID 966-PNPLA3 oligonucleotide 95 guide (antisense) chain UUAACGCAUGCUGAUGUAUUGG

[1192] Sequence ID 967-PNPLA3 oligonucleotide 96 passenger (sense) chain UGCGUUAAUUCAGCUGGUUAGCAGCCCGAAAGGCUGC

[1193] Sequence ID 968-PNPLA3 oligonucleotide 96 guide (antisense) chain UAACCAGCUGAAUUAACGCAGG

[1194] Sequence ID 969-PNPLA3 oligonucleotide 97 passenger (sense) chain GCGUUAAUUCAGCUGGUUGAGCAGCCGAAAGGCUGC

[1195] Sequence ID 970-PNPLA3 oligonucleotide 97 guide (antisense) chain UCAACCAGCUGAAUUAACGCGG

[1196] Sequence ID 971-PNPLA3 oligonucleotide 98 passenger (sense) chain CGUUAAUUCAGCUGGUUGGAGCAGCCGAAAGGCUGC

[1197] Sequence ID 972-PNPLA3 oligonucleotide 98 guide (antisense) chain UCCAACCAGCUGAAUUAACGGG

[1198] Sequence ID 973-PNPLA3 oligonucleotide 99 passenger (sense) chain UUAAUUCAGCUGGUUGGGAAGCAGCCGAAAGGCUGC

[1199] Sequence ID 974-PNPLA3 oligonucleotide 99 guide (antisense) chain UUCCCAACCAGCUGAAUUAAGG

[1200] Sequence ID 975-PNPLA3 oligonucleotide 100 passenger (sense) chain UAAUUCAGCUGGUUGGGAAAGCAGCCGAAAGGCUGC

[1201] Sequence ID 976-PNPLA3 oligonucleotide 100 guide (antisense) chain UUUCCCAACCAGCUGAAUUAGG

[1202] Sequence ID 977-PNPLA3 oligonucleotide 101 passenger (sense) chain GCAGAGGGUCCCUUACUGAAGCAGCCGAAAGGCUGC

[1203] Sequence ID 978-PNPLA3 oligonucleotide 101 guide (antisense) chain UUCAGUAAGGGACCCUCUGCGG

[1204] Sequence ID 979-PNPLA3 oligonucleotide 102 passenger (sense) chain CUAUUAAUGGUCAGACUGUAGCAGCCGAAAGGCUGC

[1205] Sequence ID 980-PNPLA3 oligonucleotide 102 guide (antisense) chain UACAGUCUGACCAUUAAUAGGG

[1206] Sequence ID 981-PNPLA3 oligonucleotide 103 passenger (sense) chain UAUUAAUGGUCAGACUGUUAGCAGCCCGAAAGGCUGC

[1207] Sequence ID 982-PNPLA3 oligonucleotide 103 guide (antisense) chain UAACAGUCUGACCAUUAAUAGG

[1208] Sequence ID 983-PNPLA3 oligonucleotide 104 passenger (sense) chain AUUAAUGGUCAGACUGUUCAGCAGCCGAAAGGCUGC

[1209] Sequence ID 984-PNPLA3 oligonucleotide 104 guide (antisense) chain UGAACAGUCUGACCAUUAAUGG

[1210] Sequence ID 985-PNPLA3 oligonucleotide 105 passenger (sense) chain UUAAUGGUCAGACUGUUCCAGCAGCCCGAAAGGCUGC

[1211] Sequence ID 986-PNPLA3 oligonucleotide 105 guide (antisense) chain UGGAACAGUCUGACCAUUAAGG

[1212] Sequence ID 987-PNPLA3 oligonucleotide 106 passenger (sense) chain UAAUGGUCAGACUGUUCCAAGCAGCCGAAAGGCUGC

[1213] Sequence ID 988-PNPLA3 oligonucleotide 106 guide (antisense) chain UUGGAACAGUCUGACCAUUAGG

[1214] Sequence ID 989-PNPLA3 oligonucleotide 107 passenger (sense) chain AAUGGUCAGACUGUUCCAGAGCAGCCGAAAGGCUGC

[1215] Sequence ID 990-PNPLA3 oligonucleotide 107 guide (antisense) chain UCUGGAACAGUCUGACCAUUGG

[1216] Sequence ID 991-PNPLA3 oligonucleotide 108 passenger (sense) chain AGAACACCUUUUUCACCUAAGCAGCCGAAAGGCUGC

[1217] Sequence ID 992-PNPLA3 oligonucleotide 108 guide (antisense) chain UUAGGUGAAAAAGGUGUUCUGG

[1218] Sequence ID 993-PNPLA3 oligonucleotide 109 passenger (sense) chain GAACACCUUUUUCACCUAAAGCAGCCGAAAGGCUGC

[1219] Sequence ID 994-PNPLA3 oligonucleotide 109 guide (antisense) chain UUUAGGUGAAAAAGGUGUUCGG

[1220] Sequence ID 995-PNPLA3 oligonucleotide 110 passenger (sense) chain AACACCUUUUUCACCUAACAGCAGCCCGAAAGGCUGC

[1221] Sequence ID 996-PNPLA3 oligonucleotide 110 guide (antisense) chain UGUUAGGUGAAAAAGGUGUUGG

[1222] Sequence ID 997-PNPLA3 oligonucleotide 111 passenger (sense) chain ACACCUUUUUCACCUAACUAGCAGCCGAAAGGCUGC

[1223] Sequence ID 998-PNPLA3 oligonucleotide 111 guide (antisense) chain UAGUUAGGUGAAAAAGGUGUGG

[1224] Sequence ID 999-PNPLA3 oligonucleotide 112 passenger (sense) chain CACCUUUUUCACCUAACUAAGCAGCCGAAAGGCUGC

[1225] Sequence ID 1000-PNPLA3 oligonucleotide 112 guide (antisense) chain UUAGUUAGGUGAAAAAGGUGGG

[1226] Sequence ID 1001-PNPLA3 oligonucleotide 113 passenger (sense) chain ACCUUUUUCACCUAACUAAAGCAGCCCGAAAGGCUGC

[1227] Sequence ID 1002-PNPLA3 oligonucleotide 113 guide (antisense) chain UUUAGUUAGGUGAAAAAGGUGG

[1228] Sequence ID 1003-PNPLA3 oligonucleotide 114 passenger (sense) chain CUUUUUCACCUAACUAAAAAGCAGCCGAAAGGCUGC

[1229] Sequence ID 1004-PNPLA3 oligonucleotide 114 guide (antisense) chain UUUUUAGUUAGGUGAAAAAGGG

[1230] Sequence ID 1005-PNPLA3 oligonucleotide 115 passenger (sense) chain UUUUCACCUAACUAAAAUAAGCAGCCGAAAGGCUGC

[1231] Sequence ID 1006-PNPLA3 oligonucleotide 115 guide (antisense) chain UUAUUUUAGUUAGGUGAAAAGG

[1232] Sequence ID 1007-PNPLA3 oligonucleotide 116 passenger (sense) chain UUCACCUAACUAAAAUAAUAGCAGCCGAAAGGCUGC

[1233] Sequence ID 1008-PNPLA3 oligonucleotide 116 guide (antisense) chain UAUUAUUUUAGUUAGGUGAAGG

[1234] Sequence ID 1009-PNPLA3 oligonucleotide 117 passenger (sense) chain UCACCUAACUAAAAUAAUGAGCAGCCGAAAGGCUGC

[1235] Sequence ID 1010-PNPLA3 oligonucleotide 117 guide (antisense) chain UCAUUAUUUUAGUUAGGUGAGG

[1236] Sequence ID 1011-PNPLA3 oligonucleotide 118 passenger (sense) chain CACCUAACUAAAAUAAUGUAGCAGCCGAAAGGCUGC

[1237] Sequence ID 1012-PNPLA3 oligonucleotide 118 guide (antisense) chain UACAUUAUUUUAGUUAGGUGGG

[1238] Sequence ID 1013-PNPLA3 oligonucleotide 119 passenger (sense) chain ACCUAACUAAAAUAAUGUUAGCAGCCCGAAAGGCUGC

[1239] Sequence ID 1014-PNPLA3 oligonucleotide 119 guide (antisense) chain UAACAUUAUUUUAGUUAGGUGG

[1240] Sequence ID 1015-PNPLA3 oligonucleotide 120 passenger (sense) chain CCUAACUAAAAUAAUGUUUAGCAGCCGAAAGGCUGC

[1241] Sequence ID 1016-PNPLA3 oligonucleotide 120 guide (antisense) chain UAAACAUUAUUUUAGUUAGGGG

[1242] Sequence ID 1017-PNPLA3 oligonucleotide 121 passenger (sense) chain CUAACUAAAAUAAUGUUUAAGCAGCCGAAAGGCUGC

[1243] Sequence ID 1018-PNPLA3 oligonucleotide 121 guide (antisense) chain UUAAACAUUAUUUUAGUUAGGG

[1244] Sequence ID 1019-PNPLA3 oligonucleotide 122 passenger (sense) chain UAACUAAAAAAAUGUUUUAAAGCAGCCCGAAAGGCUGC

[1245] Sequence ID 1020-PNPLA3 oligonucleotide-122 guide (antisense) chain UUUAAACAUUAUUUUAGUUAGG

[1246] Sequence ID 1021-PNPLA3 oligonucleotide 123 passenger (sense) chain AACUAAAAUAAUGUUUAAAAGCAGCCCGAAAGGCUGC

[1247] Sequence ID 1022-PNPLA3 oligonucleotide 123 guide (antisense) chain UUUUAAACAUUAUUUUAGUUGG

[1248] Sequence ID 1023-PNPLA3 oligonucleotide 124 passenger (sense) chain ACUAAAAUAAUGUUUAAAGAGCAGCCGAAAGGCUGC

[1249] Sequence ID 1024-PNPLA3 oligonucleotide-124 guide (antisense) chain UCUUUAAACAUUAUUUUAGUGG

[1250] Sequence ID 1025-PNPLA3 oligonucleotide 125 passenger (sense) chain CUAAAAUAAUGUUUAAAGAAGCAGCCGAAAGGCUGC

[1251] Sequence ID 1026-PNPLA3 oligonucleotide 125 guide (antisense) chain UUCUUUAAACAUUAUUUUAGGG

[1252] Sequence ID 1027-PNPLA3 oligonucleotide 126 passenger (sense) chain UAAAAUAAUGUUUAAAGAGAGAGCAGCCGAAAGGCUGC

[1253] Sequence ID 1028-PNPLA3 oligonucleotide 126 guide (antisense) chain UCUCUUUAAACAUUAUUUUAGG

[1254] Sequence ID 1029-PNPLA3 oligonucleotide 127 passenger (sense) chain AAGAGUUUUGUAUAAAAAUAGCAGCCGAAAGGCUGC

[1255] Sequence ID 1030-PNPLA3 oligonucleotide 127 guide (antisense) chain UAUUUUUAUACAAAACUCUUGG

[1256] Sequence ID 1031-PNPLA3 oligonucleotide 128 passenger (sense) chain GUUUUGUAUAAAAAUGUAAAGCAGCCGAAAGGCUGC

[1257] Sequence ID 1032-PNPLA3 oligonucleotide 128 guide (antisense) chain UUUACAUUUUUUAUACAAAAC Role

[1258] Sequence ID 1033-PNPLA3 oligonucleotide 129 passenger (sense) chain UUGUAUAAAAAUGUAAGGAAGCAGCCGAAAGGCUGC

[1259] Sequence ID 1034-PNPLA3 oligonucleotide 129 guide (antisense) chain UUCCUUACAUUUUUAUACAAGG

[1260] Sequence ID 1035-PNPLA3 oligonucleotide 130 passenger (sense) chain UGUAUAAAAAUGUAAGGAAAGCAGCCGAAAGGCUGC

[1261] Sequence ID 1036-PNPLA3 oligonucleotide 130 guide (antisense) chain UUUCCUUACAUUUUUAUACAGG

[1262] Sequence ID 1037-PNPLA3 oligonucleotide 131 passenger (sense) chain GUAUAAAAAAUGUAAGGAAGAGCAGCCGAAAGGCUGC

[1263] Sequence ID 1038-PNPLA3 oligonucleotide 131 guide (antisense) chain UCUUCCUUACAUUUUUAUACGG

[1264] Sequence ID 1039-PNPLA3 oligonucleotide 132 passenger (sense) chain AUGUAAGGAAGCGUUGUUAAGCAGCCGAAAGGCUGC

[1265] Sequence ID 1040-PNPLA3 oligonucleotide-132 guide (antisense) chain UUAACAACGCUUCCUUACAUGG

[1266] Sequence ID 1041-PNPLA3 oligonucleotide 133 passenger (sense) chain UUUUGUAUUAUGUGAAUCAAGCAGCCGAAAGGCUGC

[1267] Sequence ID 1042-PNPLA3 oligonucleotide 133 guide (antisense) chain UUGAUUCACAUAAUACAAAAGG

[1268] Sequence ID 1043-PNPLA3 oligonucleotide 134 passenger (sense) chain UUUGUAUUAUGUGAAUCAGAGCAGCCGAAAGGCUGC

[1269] Sequence ID 1044-PNPLA3 oligonucleotide-134 guide (antisense) chain UCUGAUUCACAUAAUACAAAGG

[1270] Sequence ID 1045-PNPLA3 oligonucleotide 135 passenger (sense) chain UUGUAUUAUGUGAAUCAGUAGCAGCCGAAAGGCUGC

[1271] Sequence ID 1046-PNPLA3 oligonucleotide 135 guide (antisense) chain UACUGAUUCACAUAAUACAAGG

[1272] Sequence ID 1047-PNPLA3 oligonucleotide 136 passenger (sense) chain GUAUUAUGUGAAUCAGUGAAGCAGCCGAAAGGCUGC

[1273] Sequence ID 1048-PNPLA3 oligonucleotide 136 guide (antisense) chain UUCACUGAUUCACAUAAUACGG

[1274] Sequence ID 1049-PNPLA3 oligonucleotide 137 passenger (sense) chain UAUUAUGUGAAUCAGUGAGAGCAGCCGAAAGGCUGC

[1275] Sequence ID 1050-PNPLA3 oligonucleotide 137 guide (antisense) chain UCUCACUGAUUCACAUAAUAGG

[1276] Sequence ID 1051-PNPLA3 oligonucleotide 138 passenger (sense) chain AUUAUGUGAAUCAGUGAGAAGCAGCCGAAAGGCUGC

[1277] Sequence ID 1052-PNPLA3 oligonucleotide 138 guide (antisense) chain UUCUCACUGAUUCACAUAAUGG

[1278] Sequence ID 1053-PNPLA3 oligonucleotide 139 passenger (sense) chain UUAUGUGAAUCAGUGAGAUAGCAGCCGAAAGGCUGC

[1279] Sequence ID 1054-PNPLA3 oligonucleotide 139 guide (antisense) chain UAUCUCACUGAUUCACAUAAGG

[1280] Sequence ID 1055-PNPLA3 oligonucleotide 140 passenger (sense) chain UAUGUGAAUCAGUGAGAUGAGCAGCCGAAAGGCUGC

[1281] Sequence ID 1056-PNPLA3 oligonucleotide 140 guide (antisense) chain UCAUCUCACUGAUUCACAUAGG

[1282] Sequence ID 1057-PNPLA3 oligonucleotide 141 passenger (sense) chain AUGUGAAUCAGUGAGAUGUAGCAGCCGAAAGGCUGC

[1283] Sequence ID 1058-PNPLA3 oligonucleotide 141 guide (antisense) chain UACAUCUCACUGAUUCACAUGG

[1284] Sequence ID 1059-PNPLA3 oligonucleotide 142 passenger (sense) chain UGUGAAUCAGUGAGAUGUUAGCAGCCCGAAAGGCUGC

[1285] Sequence ID 1060-PNPLA3 oligonucleotide 142 guide (antisense) chain UAACAUCUCACUGAUUCACAGG

[1286] Sequence ID 1061-PNPLA3 oligonucleotide 143 passenger (sense) chain GUGAAUCAGUGAGAUGUUAAGCAGCCGAAAGGCUGC

[1287] Sequence ID 1062-PNPLA3 oligonucleotide 143 guide (antisense) chain UUAACAUCUCACUGAUUCACGG

[1288] Sequence ID 1063-PNPLA3 oligonucleotide 144 passenger (sense) chain GUGAGAUGUUAGUAGAAUAAGCAGCCGAAAGGCUGC

[1289] Sequence ID 1064-PNPLA3 oligonucleotide 144 guide (antisense) chain UUAUUCUACUAACAUCUCACGG

[1290] Sequence ID 1065-PNPLA3 oligonucleotide 145 passenger (sense) chain UUUCUAUUUAUGCAUUUGAAGCAGCCCGAAAGGCUGC

[1291] Sequence ID 1066-PNPLA3 oligonucleotide 145 guide (antisense) chain UUCAAAUGCAUAAAUAGAAAGG

[1292] Sequence ID 1067-PNPLA3 oligonucleotide 146 passenger (sense) chain CUAUUUAUGCAUUUGAGUAAGCAGCCGAAAGGCUGC

[1293] Sequence ID 1068-PNPLA3 oligonucleotide 146 guide (antisense) chain UUACUCAAAUGCAUAAAUAGGG

[1294] Sequence ID 1069-PNPLA3 oligonucleotide 147 passenger (sense) chain UGCUCAAACUGUUAAAUGUAGCAGCCCGAAAGGCUGC

[1295] Sequence ID 1070-PNPLA3 oligonucleotide 147 guide (antisense) chain UACAUUUAACAGUUUGAGCAGG

[1296] Sequence ID 1071-PNPLA3 oligonucleotide 148 passenger (sense) chain GCUCAAACUGUUAAAUGUUAGCAGCCGAAAGGCUGC

[1297] Sequence ID 1072-PNPLA3 oligonucleotide 148 guide (antisense) chain UAACAUUUAACAGUUUGAGCGG

[1298] Sequence ID 1073-PNPLA3 oligonucleotide 149 passenger (sense) chain CUCAAACUGUUAAAUGUUGAGCAGCCGAAAGGCUGC

[1299] Sequence ID 1074-PNPLA3 oligonucleotide 149 guide (antisense) chain UCAACAUUUAACAGUUUGAGGG

[1300] Sequence ID 1075-PNPLA3 oligonucleotide 150 passenger (sense) chain UCAAACUGUUAAAUGUUGGAGCAGCCGAAAGGCUGC

[1301] Sequence ID 1076-PNPLA3 oligonucleotide 150 guide (antisense) chain UCCAACAUUUAACAGUUUGAGG

[1302] Sequence ID 1077-PNPLA3 oligonucleotide 151 passenger (sense) chain CAAACUGUUAAAUGUUGGAAGCAGCCGAAAGGCUGC

[1303] Sequence ID 1078-PNPLA3 oligonucleotide 151 guide (antisense) chain UUCCAACAUUUAACAGUUUGGG

[1304] Sequence ID 1079-PNPLA3 oligonucleotide 152 passenger (sense) chain AAACUGUUAAAUGUUGGAAAGCAGCCGAAAGGCUGC

[1305] Sequence ID 1080-PNPLA3 oligonucleotide 152 guide (antisense) chain UUUCCAACAUUUAACAGUUUGG

[1306] Sequence ID 1081-PNPLA3 oligonucleotide 153 passenger (sense) chain AACUGUUAAAUGUUGGAAAAGCAGCCGAAAGGCUGC

[1307] Sequence ID 1082-PNPLA3 oligonucleotide 153 guide (antisense) chain UUUUCCAACAUUUAACAGUUGG

[1308] Sequence ID 1083-PNPLA3 oligonucleotide 154 passenger (sense) chain UGUUAAAUGUUGGAAAAGAAGCAGCCGAAAGGCUGC

[1309] Sequence ID 1084-PNPLA3 oligonucleotide 154 guide (antisense) chain UUCUUUUCCAACAUUUAACAGG

[1310] Sequence ID 1085-PNPLA3 oligonucleotide 155 passenger (sense) chain GUUAAAUGUUGGAAAAGAAAGCAGCCCGAAAGGCUGC

[1311] Sequence I...

Claims

1. An RNAi oligonucleotide comprising a sense strand and an antisense strand, wherein the sense strand and the antisense strand form a double-stranded region. The aforementioned sense chain, [mCs][mU][mA][mA][mC][mU][mA][fA][fA][fA][fU][mA][mA][mU][mG][mU][mU][mU][mA][mA][mG][mC][mA][mG][mC][mC][mG][ademA-GalN Ac][ademA-GalN Ac][ademA-GalNAc][mG][mG][mC][mU][mG][mC] (Sequence ID 1250) And, Here, the creotides at positions 1, 2, 3, 4, 5, 6, 7, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 31, 32, 33, 34, 35 and 36 are modified with 2'-O-methyl, the nucleotides at positions 8, 9, 10 and 11 are modified with 2'-fluoro, the internucleotide linkage between the nucleotides at positions 1 and 2 is a phosphorothioate bond, and the nucleotides at positions 28, 29 and 30 are modified with adem-GalNAc. The aforementioned antisense chain, [MePhosphonate-4O-mUs][fUs][fAs][fA][fA][mC][fA][mU][mU][fA][mU][mU][mU][fU][mA][mG][mU][mU][mA][mGs][mGs][mG] (SEQ ID NO: 1251) And, Herein, the nucleotides at positions 1, 6, 8, 9, 11, 12, 13, 15, 16, 17, 18, 19, 20, 21, and 22 are modified with 2'-O-methyl, the nucleotides at positions 2, 3, 4, 5, 7, 10, and 14 are modified with 2'-fluoro, the internucleotide links between positions 1 and 2, 2 and 3, 3 and 4, 20 and 21, and 21 and 22 are phosphorothioate links, and the nucleotide at position 1 is a 4'-phosphate analog containing 5'-methoxyphosphonate-4'-oxy.

2. A pharmaceutical composition comprising an RNAi oligonucleotide, wherein the RNAi oligonucleotide comprises a sense strand and an antisense strand, and the sense strand and antisense strand form a double-stranded region. The aforementioned sense chain, [mCs][mU][mA][mA][mC][mU][mA][fA][fA][fA][fU][mA][mA][mU][mG][mU][mU][mU][mA][mA][mG][mC][mA][mG][mC][mC][mG][ademA-GalN Ac][ademA-GalN Ac][ademA-GalNAc][mG][mG][mC][mU][mG][mC] (Sequence ID 1250) And, Here, the creotides at positions 1, 2, 3, 4, 5, 6, 7, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 31, 32, 33, 34, 35 and 36 are modified with 2'-O-methyl, the nucleotides at positions 8, 9, 10 and 11 are modified with 2'-fluoro, the internucleotide linkage between the nucleotides at positions 1 and 2 is a phosphorothioate bond, and the nucleotides at positions 28, 29 and 30 are modified with adem-GalNAc. The aforementioned antisense chain, [MePhosphonate-4O-mUs][fUs][fAs][fA][fA][mC][fA][mU][mU][fA][mU][mU][mU][fU][mA][mG][mU][mU][mA][mGs][mGs][mG] (SEQ ID NO: 1251) And, Herein, the nucleotides at positions 1, 6, 8, 9, 11, 12, 13, 15, 16, 17, 18, 19, 20, 21, and 22 are modified with 2'-O-methyl, the nucleotides at positions 2, 3, 4, 5, 7, 10, and 14 are modified with 2'-fluoro, the internucleotide links between positions 1 and 2, 2 and 3, 3 and 4, 20 and 21, and 21 and 22 are phosphorothioate bonds, and the nucleotide at position 1 is a 4'-phosphate analog containing 5'-methoxyphosphonate-4'-oxy.

3. The pharmaceutical composition according to claim 2, further comprising a carrier suitable for intravenous administration.

4. The pharmaceutical composition according to claim 3, wherein the carrier contains water.

5. The pharmaceutical composition according to claim 3, wherein the carrier comprises phosphate-buffered saline.

6. An RNAi oligonucleotide according to claim 1, or a pharmaceutical composition according to any one of claims 2 to 5, for use in reducing PNPLA3 expression in cells, a population of cells, or a subject.

7. The RNAi oligonucleotide or pharmaceutical composition according to claim 6, wherein reducing PNPLA3 expression includes reducing the amount or level of PNPLA3 mRNA, the amount or level of PNPLA3 protein, or both.

8. The RNAi oligonucleotide or pharmaceutical composition according to claim 6, wherein the subject has a disease, disorder, or condition related to PNPLA3 expression.

9. The RNAi oligonucleotide or pharmaceutical composition according to claim 8, wherein the disease, disorder, or condition associated with PNPLA3 expression is a cardiometabolic disease, alcoholic hepatitis (AH), alcoholic liver disease (ALD), cirrhosis, hepatocellular carcinoma (HCC), non-alcoholic fatty liver disease (NAFLD), or non-alcoholic steatohepatitis (NASH).

10. An RNAi oligonucleotide according to claim 1, or a pharmaceutical composition according to any one of claims 2 to 5, for use in treating subjects having cardiometabolic disease, alcoholic hepatitis (AH), alcoholic liver disease (ALD), cirrhosis, hepatocellular carcinoma (HCC), non-alcoholic fatty liver disease (NAFLD), or non-alcoholic steatohepatitis (NASH).

11. The RNAi oligonucleotide or pharmaceutical composition according to claim 6, administered in combination with a second composition or therapeutic agent.

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