UNC13A antisense oligonucleotides and uses thereof

Abstract

The present disclosure relates, in some aspects, to compositions of UNC13A antisense oligonucleotides (ASOs) and methods of using ASO to alter UNC13A concealment exon splicing and increase UNC13A protein expression. Such ASO may be used in the treatment of neurodegenerative disorders, in particular neurodegenerative disorders associated with the TAR-DNA binding protein-43 (TDP-43) pathology, such as amyotrophic lateral sclerosis (ALS) or frontotemporal dementia (FTD).

Description

[0001] Cross - Reference to Related Applications

[0002] This application claims priority to U.S. Provisional Application No. 63 / 378,473, filed Oct. 5, 2022, the content of which is incorporated herein by reference in its entirety.

[0003] Sequence Listing Submitted as an ASCII Text File

[0004] The content of the electronic sequence listing (792752001540seqlist.xml; size: 1,083,815 bytes; and creation date: Oct. 4, 2023) is incorporated herein by reference in its entirety. TECHNICAL FIELD

[0005] In some aspects, the present disclosure relates to compositions and methods for increasing UNC13A protein expression using antisense oligonucleotides that target the cryptic exon locus of UNC13A. BACKGROUND OF THE INVENTION

[0006] The hallmark pathological feature of neurodegenerative diseases, amyotrophic lateral sclerosis (ALS) and frontotemporal dementia (FTD), is the depletion of the RNA-binding protein TDP-43 from the nuclei of neurons in the brain and spinal cord. TDP-43, encoded by TARDBP, is an abundant ubiquitously expressed RNA-binding protein that is normally localized to the nucleus. It plays a role in essential RNA processing activities including RNA transcription, alternative splicing, and RNA transport (1). TDP-43 can bind to thousands of pre-messenger RNA / mRNA targets (2, 3). The reduction of TDP-43 in other normal adult nervous systems alters the splicing or expression levels of over 1,500 RNAs, including transcripts with long introns (2). The major splicing regulatory function of TDP-43 is to inhibit the inclusion of cryptic exons during splicing (4-7). Unlike normal conserved exons, these cryptic exons lurk within introns and are typically excluded from mature mRNA. When TDP-43 is depleted from cells, these cryptic exons are spliced into messenger RNA, usually introducing frameshifts and premature terminations or even nonsense-mediated decay of the mRNA. However, there remains a need to identify disease-critical cryptic splicing events. Thus, there is a need to discover cryptic splicing targets that are regulated by TDP-43 and also play a role in the pathogenesis of TDP-43 proteinopathies as therapeutic targets. The present disclosure provides methods and compositions to address such and other needs. SUMMARY OF THE INVENTION

[0007] The present disclosure provides antisense oligonucleotides (ASOs) that modulate UNC13A splicing, wherein the ASO sequences are complementary to the sequence of UNC13A mRNA, which sequence is located between the 5' splice donor site and the 3' splice acceptor site of a cryptic exon located between exons 20 and 21 of the UNC13A gene. The ASOs provided herein have a length between 13 and 30 nucleotides. Modified ASOs containing one or more modified nucleotides are also provided, such as 2'-OMe antisense oligonucleotides, 2'-O-methoxyethyl antisense oligonucleotides, phosphorothioate antisense oligonucleotides, and LNA antisense oligonucleotides. Methods of using the described ASOs are also provided. In some aspects, a method for increasing the expression of full-length UNC13A protein in a cell by at least 10% is provided. In other aspects, a method for treating a clinical disorder in an individual, such as amyotrophic lateral sclerosis (ALS) or frontotemporal dementia (FTD), is provided. Pharmaceutical compositions and kits containing one or more of the ASOs described herein are also provided.

[0008] The present disclosure provides a method for increasing the expression of UNC13A protein in a cell by at least 10%, the method comprising contacting the cell with an antisense oligonucleotide comprising 13 to 30 nucleotides complementary to the sequence shown in SEQ ID NO:1.

[0009] The present disclosure provides a method for increasing the expression of UNC13A protein in a cell by at least 10%, the method comprising contacting the cell with an antisense oligonucleotide comprising 13 to 30 nucleotides, wherein the antisense oligonucleotide is complementary to the sequence shown in SEQ ID NO:1. The present disclosure also provides a method for increasing the expression of UNC13A protein in a cell by at least 10%, the method comprising contacting the cell with an antisense oligonucleotide comprising 13 to 30 nucleotides, wherein the antisense oligonucleotide is complementary to the sequence or a portion thereof shown in any one of SEQ ID NOs: 1339 - 1342. In some embodiments, the antisense oligonucleotide comprises the sequence shown in any one of SEQ ID NOs: 20 - 355, 400 - 638, 810 - 957, or 961 - 1332.

[0010] In some embodiments, the antisense oligonucleotide consists of 13 to 25 nucleotides, wherein the antisense oligonucleotide is complementary to the sequence shown in SEQ ID NO:1. In some embodiments, the antisense oligonucleotide consists of 13, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24 or 25 nucleotides, wherein the antisense oligonucleotide is complementary to the sequence shown in SEQ ID NO:1. In some embodiments, the antisense oligonucleotide consists of 19, 20, 21 or 22 nucleotides, wherein the antisense oligonucleotide is complementary to the sequence shown in SEQ ID NO:1.

[0011] In some embodiments, the antisense oligonucleotide consists of 13 nucleotides complementary to the sequence shown in SEQ ID NO:1, and wherein the antisense oligonucleotide consists of the sequence shown in any one of SEQ ID NOs: 961-983 and 1116-1145. In some embodiments, the antisense oligonucleotide consists of 15 nucleotides complementary to the sequence shown in SEQ ID NO:1, and wherein the antisense oligonucleotide consists of the sequence shown in any one of SEQ ID NOs: 984-1004 and 1146-1173. In some embodiments, the antisense oligonucleotide consists of 16 nucleotides complementary to the sequence shown in SEQ ID NO:1, and wherein the antisense oligonucleotide consists of the sequence shown in any one of SEQ ID NOs: 41, 42, 56, 57, 71, 72, 86, 87, 101, 102, 116, 117, 131, 132, 146, 147, 161, 162, 176, 177, 191, 192, 206, 207, 221, 222, 236, 237, 251, 252, 266, 267, 281, 282, 296, 297, 311, 326, 341, 810, 811, 812, 813, 826, 827, 835, 838, 841, 842, 851, 852, 861, 864, 867, 868, 877, 878, 895, 900, 901, 910, 934 and 935. In some embodiments, the antisense oligonucleotide consists of 17 nucleotides complementary to the sequence shown in SEQ ID NO:1, and wherein the antisense oligonucleotide consists of the sequence shown in any one of SEQ ID NOs: 43, 58, 73, 88, 103, 118, 133, 148, 163, 178, 193, 208, 223, 238, 253, 268, 283, 298, 312, 313, 327, 328, 342, 343, 814, 815, 816, 828, 829, 836, 839, 843, 844, 853, 854, 862, 865, 869, 870, 879, 880, 881, 891, 896, 897, 902, 903, 1005-1023 and 1174-1199.In some embodiments, the antisense oligonucleotide consists of 18 nucleotides complementary to the sequence shown in SEQ ID NO:1, and wherein the antisense oligonucleotide consists of the sequence shown in any one of SEQ ID NOs: 38-40, 44, 45, 59, 60, 74, 75, 89, 90, 104, 105, 119, 120, 134, 135, 149, 150, 164, 165, 179, 180, 194, 195, 209, 210, 224, 225, 239, 240, 254, 255, 269, 270, 284, 285, 299, 300, 314, 329, 344, 518-638, 817, 818, 845, 855, 871, 882, 904, 958, 959, 960, 1024-1041, and 1200-1224. In some embodiments, the antisense oligonucleotide consists of 19 nucleotides complementary to the sequence shown in SEQ ID NO:1, and wherein the antisense oligonucleotide consists of the sequence shown in any one of SEQ ID NOs: 46, 61, 76, 91, 106, 121, 136, 151, 166, 181, 196, 211, 226, 241, 256, 271, 286, 301, 315, 316, 330, 331, 345, 346, 819, 830, 846, 856, 872, 883, 884, 892, 905, 1042-1058, and 1225-1247. In some embodiments, the antisense oligonucleotide consists of 20 nucleotides complementary to the sequence shown in SEQ ID NO:1, and wherein the antisense oligonucleotide consists of the sequence shown in any one of SEQ ID NOs: 47, 48, 62, 63, 77, 78, 92, 93, 107, 108, 122, 123, 137, 138, 152, 153, 167, 168, 182, 183, 197, 198, 212, 213, 227, 228, 242, 243, 257, 258, 272, 273, 287, 288, 302, 303, 317, 332, 347, 911, 936, 937, 1059-1074, and 1248-1270.In some embodiments, the antisense oligonucleotide consists of 21 nucleotides complementary to the sequence shown in SEQ ID NO:1, and wherein the antisense oligonucleotide consists of the sequence shown in any one of SEQ ID NOs: 20 - 37, 49, 64, 79, 94, 109, 124, 139, 154, 169, 184, 199, 214, 229, 244, 259, 274, 289, 304, 318, 319, 333, 334, 348, 349, 400 - 517, 1075 - 1089 and 1271 - 1292. In some embodiments, the antisense oligonucleotide consists of 22 nucleotides complementary to the sequence shown in SEQ ID NO:1, and wherein the antisense oligonucleotide consists of the sequence shown in any one of SEQ ID NOs: 50, 51, 65, 66, 80, 81, 95, 96, 110, 111, 125, 126, 140, 141, 155, 156, 170, 171, 185, 186, 200, 201, 215, 216, 230, 231, 245, 246, 260, 261, 275, 276, 290, 291, 305, 306, 320, 335, 350, 912, 913, 914, 938, 939, 940, 941, 1090 - 1103 and 1293 - 1313. In some embodiments, the antisense oligonucleotide consists of 23 nucleotides complementary to the sequence shown in SEQ ID NO:1, and wherein the antisense oligonucleotide consists of the sequence shown in any one of SEQ ID NOs: 52, 67, 82, 97, 112, 127, 142, 157, 172, 187, 202, 217, 232, 247, 262, 277, 292, 307, 321, 322, 336, 337, 351, 352, 820, 831, 847, 857, 873, 885, 886, 893, 906, 915 - 918, 930 and 942 - 945.In some embodiments, the antisense oligonucleotide consists of 24 nucleotides complementary to the sequence shown in SEQ ID NO:1, and wherein the antisense oligonucleotide consists of the sequence shown in any one of SEQ ID NO:53, 54, 68, 69, 83, 84, 98, 99, 113, 114, 128, 129, 143, 144, 158, 159, 173, 174, 188, 189, 203, 204, 218, 219, 233, 234, 248, 249, 263, 264, 278, 279, 293, 294, 308, 309, 323, 338, 353, 821, 822, 832, 848, 858, 874, 887, 907, 919, 920 - 923, 931, 946 - 951, 1104 - 1115 and 1314 - 1332. In some embodiments, the antisense oligonucleotide consists of 25 nucleotides complementary to the sequence shown in SEQ ID NO:1, and wherein the antisense oligonucleotide consists of the sequence shown in any one of SEQ ID No:55, 70, 85, 100, 115, 130, 145, 160, 175, 190, 205, 220, 235, 250, 265, 280, 295, 310, 324, 325, 339, 340, 354, 355, 823, 824, 825, 833, 834, 837, 840, 849, 850, 859, 860, 863, 866, 875, 876, 888 - 890, 894, 898, 899, 908, 909, 924 - 929, 932, 933, 952 and 953 - 957.

[0012] In some embodiments, the antisense oligonucleotide of the method provided herein is complementary to the sequence shown in any one of SEQ ID NO:700 - 703. In some embodiments, the antisense oligonucleotide has at least 80%, at least 90%, at least 95%, at least 98% or 100% complementarity with the sequence shown in any one of SEQ ID NO:700 - 703. In some embodiments, the antisense oligonucleotide has at least 80%, at least 90%, at least 95%, at least 98% or 100% complementarity with the sequence shown in SEQ ID NO:701. In some embodiments, the antisense oligonucleotide has at least 80%, at least 90%, at least 95%, at least 98% or 100% complementarity with the sequence shown in SEQ ID NO:703.

[0013] In some embodiments, the antisense oligonucleotide of the method provided herein is complementary to the sequence shown in any one of SEQ ID NOs: 1339-1342 or a portion thereof. In some embodiments, the antisense oligonucleotide is complementary to the sequence shown in SEQ ID NO: 1339 or a portion thereof. In some embodiments, the antisense oligonucleotide is complementary to the sequence shown in SEQ ID NO: 1340 or a portion thereof. In some embodiments, the antisense oligonucleotide is complementary to the sequence shown in SEQ ID NO: 1341 or a portion thereof. In some embodiments, the antisense oligonucleotide is complementary to the sequence shown in SEQ ID NO: 1342 or a portion thereof. In some embodiments, the antisense oligonucleotide is complementary to at least 1, at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, at least 11, at least 12, or at least 13 nucleotides of the sequence shown in any one of SEQ ID NOs: 1339-1342. In some embodiments, the antisense oligonucleotide is complementary to at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, at least 11, at least 12, or at least 13 consecutive nucleotides of the sequence shown in any one of SEQ ID NOs: 1339-1342. In some embodiments, the antisense oligonucleotide is at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99%, or 100% complementary to the sequence shown in SEQ ID NO: 1339. In some embodiments, the antisense oligonucleotide is at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99%, or 100% complementary to the sequence shown in SEQ ID NO: 1340. In some embodiments, the antisense oligonucleotide is at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99%, or 100% complementary to the sequence shown in SEQ ID NO: 1341. In some embodiments, the antisense oligonucleotide is at least at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99%, or 100% complementary to the sequence shown in SEQ ID NO: 1342.

[0014] In some embodiments, prior to contacting the cell with the antisense oligonucleotide, the cell expresses a low level of UNC13A. In some embodiments, it includes measuring the level of UNC13A protein in the cell. In some embodiments, the level of UNC13A protein is measured by Western blot, imaging, ELISA, fluorescence reporter assay, luminescence assay, or immunohistochemistry.

[0015] In some embodiments, the secondary structure of UNC13A mRNA is altered by the binding of the antisense oligonucleotide to the UNC13A transcript. In some embodiments, the antisense oligonucleotide sterically blocks the inclusion of the UNC13A cryptic exon. In some embodiments, the antisense oligonucleotide has at least 80%, at least 90%, at least 95%, at least 98% or 100% complementarity with the sequence shown in SEQ ID NO:1.

[0016] In some embodiments, the antisense oligonucleotide of the methods provided herein restores the expression of UNC13A mRNA in cells to at least 25%, at least 30%, at least 35%, at least 40%, at least 45% or at least 50% of the level of UNC13A mRNA expressed in wild-type cells. In some embodiments, the antisense oligonucleotide increases the expression of UNC13A mRNA in cells by at least 50%, at least 100%, at least 200%, at least 300%, at least 400% or at least 500%. In some embodiments, the antisense oligonucleotide consists of the sequence shown in any one of SEQ ID NO:1059 - 1074, 1075 - 1089, 1090 - 1103, 1248 - 1270, 1271 - 1292 and 1293 - 1313.

[0017] In some embodiments, the antisense oligonucleotide of the methods provided herein reduces the expression of UNC13A cryptic exon splice variant mRNA by at least 10%. In some embodiments, the antisense oligonucleotide reduces the expression of UNC13A cryptic exon mRNA by at least 10%, at least 20%, at least 30%, at least 40% or at least 50% of the level of UNC13A cryptic exon mRNA expressed in cells treated with shRNA, miRNA, antisense oligonucleotide targeting TARDBP. In some embodiments, the antisense oligonucleotide reduces the expression of UNC13A cryptic exon mRNA in cells by at least 10%, at least 20%, at least 30%, at least 40% or at least 50%. In some embodiments, the antisense oligonucleotide consists of the sequence shown in any one of SEQ ID NO:1059 - 1074, 1075 - 1089, 1090 - 1103, 1248 - 1270, 1271 - 1292 and 1293 - 1313.

[0018] In some embodiments, the UNC13A cryptic exon splice variant comprises the sequence shown in SEQ ID NO:7 or SEQ ID NO:9.

[0019] In some embodiments, after the cells are contacted with the antisense oligonucleotide, the ratio of the UNC13A transcript that does not contain the cryptic exon splicing variant to the UNC13A transcript that contains the cryptic exon splicing variant is at least 10:1. In some embodiments, the presence and / or amount of the cryptic exon splicing variant is measured using qPCR.

[0020] In some embodiments, the cells are neurons. In some embodiments, the neurons are motor neurons, cortical neurons, interneurons, or excitatory neurons.

[0021] In some embodiments, contacting the cells with the antisense oligonucleotide of the methods provided herein increases the expression of UNC13A protein to a level sufficient to improve one or more neuronal functions. In some embodiments, the one or more neuronal functions are selected from the group consisting of neuronal signaling, neuronal morphology, neurite length, neurotransmitter release, action potential, neuronal synaptic plasticity, and neuronal lifespan.

[0022] In some embodiments, the antisense oligonucleotide of the methods provided herein restores the expression of UNC13A protein in the cells to at least 10%, at least 20%, at least 30%, at least 40%, or at least 50% of the level of UNC13A protein expressed in wild-type cells. In some embodiments, the antisense oligonucleotide increases the expression of UNC13A protein in the cells by at least 50%, at least 100%, at least 200%, at least 300%, at least 400%, or at least 500%. In some embodiments, the antisense oligonucleotide consists of the sequences shown in any one of SEQ ID NO: 1059 - 1074, 1075 - 1089, 1090 - 1103, 1248 - 1270, 1271 - 1292, and 1293 - 1313.

[0023] In some embodiments, the antisense oligonucleotides of the methods provided herein increase the expression of UNC13A mRNA in cells by at least 50%, decrease the expression of UNC13A cryptic exon mRNA in the cells by at least 25%, and increase the expression of UNC13A protein in the cells by at least 50%. In some embodiments, the antisense oligonucleotides consist of the sequences shown in any one of SEQ ID NOs: 1059 - 1074, 1075 - 1089, 1090 - 1103, 1248 - 1270, 1271 - 1292, and 1293 - 1313. In some embodiments, the antisense oligonucleotides consist of the sequences shown in any one of SEQ ID NOs: 1059 - 1074, 1075 - 1089, and 1090 - 1103. In some embodiments, the antisense oligonucleotides consist of the sequences shown in any one of SEQ ID NOs: 1248 - 1270, 1271 - 1292, and 1293 - 1313.

[0024] In some embodiments, the cells of the methods provided herein are heterozygous or homozygous for a risk allele. In some embodiments, the risk allele is selected from the group consisting of: rs12608932 (hg38 chr19:17,641,880 A→C), rs12973192 (hg38 chr19:17,642,430 C→G), rs56041637 (hg38 chr19:17,642,033 - 17,642,056 0 - 2 CATC repeat→3 - 5 CATC repeat), and rs62121687 (hg38 chr19:17,642,351 C→A) or any combination thereof. In some embodiments, the cells contain a mutation associated with amyotrophic lateral sclerosis (ALS) or frontotemporal dementia (FTD).

[0025] In some embodiments, the antisense oligonucleotides are provided to the cells by transfection. In some embodiments, the cells express low levels of TDP - 43 protein. In some embodiments, the cells are human cells or murine cells.

[0026] In some embodiments, the antisense oligonucleotides of the methods provided herein do not cause any cell morphological changes. In some embodiments, the antisense oligonucleotides do not cause neuronal toxicity. In some embodiments, the antisense oligonucleotides do not bind to genomic sequences other than the sequence shown in SEQ ID NO: 1.

[0027] The present invention also provides a method of treating an individual having amyotrophic lateral sclerosis (ALS) or frontotemporal dementia (FTD), the method comprising administering to the individual an antisense oligonucleotide comprising 13 to 30 nucleotides, wherein the antisense oligonucleotide is complementary to the sequence shown in SEQ ID NO:1, and wherein the individual is homozygous for a reference allele, homozygous for a risk allele, heterozygous for a risk allele, or comprises a mutation associated with ALS or FTD. In some embodiments, the risk allele is selected from the group consisting of: rs12608932 (hg38 chr19:17,641,880A→C), rs12973192 (hg38 chr19:17,642,430C→G), rs56041637 (hg38 chr19:17,642,033-17,642,056 0-2CATC repeat→3-5CATC repeat), and rs62121687 (hg38 chr19:17,642,351C→A) or any combination thereof. In some embodiments, the mutation associated with ALS or FTD is a mutation in TDP43.

[0028] In some embodiments, the antisense oligonucleotide has a sequence complementary to the sequence of UNC13A mRNA, the sequence of UNC13A mRNA being between the 5' splice donor site and the 3' splice acceptor site of a cryptic exon located between exons 20 and 21 of the UNC13A gene. In some embodiments, the antisense oligonucleotide comprises a 2'OMe antisense oligonucleotide, a 2'-O-methoxyethyl antisense oligonucleotide, a phosphorothioate antisense oligonucleotide, or a locked nucleic acid (LNA) antisense oligonucleotide.

[0029] The present invention also provides an antisense oligonucleotide that modulates UNC13A splicing, wherein the antisense oligonucleotide has a sequence complementary to the sequence of UNC13A mRNA, the sequence of UNC13A mRNA being between the 5' splice donor site and the 3' splice acceptor site of a cryptic exon located between exons 20 and 21 of the UNC13A gene. The present invention also provides an antisense oligonucleotide comprising 13 to 30 nucleotides that modulates UNC13A splicing, wherein the antisense oligonucleotide comprises any one of the sequences shown in SEQ ID NO:20-355, 400-638, 810-959, or 961-1332.

[0030] In some embodiments, the antisense oligonucleotide is complementary to the sequence shown in SEQ ID NO:1. In some embodiments, the antisense oligonucleotide is complementary to the sequence shown in any one of SEQ ID NOs:700 - 703. In some embodiments, the antisense oligonucleotide comprises at least 80%, at least 90%, at least 95%, at least 98% or 100% complementarity to the sequence shown in any one of SEQ ID NOs:700 - 703. In some embodiments, the antisense oligonucleotide comprises at least 80%, at least 90%, at least 95%, at least 98% or 100% complementarity to the sequence shown in SEQ ID NO:701. In some embodiments, the antisense oligonucleotide comprises at least 80%, at least 90%, at least 95%, at least 98% or 100% complementarity to the sequence shown in SEQ ID NO:703.

[0031] In some embodiments, the antisense oligonucleotide is complementary to the sequence shown in any one of SEQ ID NOs: 1339-1342 or a portion thereof. In some embodiments, the antisense oligonucleotide is complementary to the sequence shown in SEQ ID NO: 1339 or a portion thereof. In some embodiments, the antisense oligonucleotide is complementary to the sequence shown in SEQ ID NO: 1340 or a portion thereof. In some embodiments, the antisense oligonucleotide is complementary to the sequence shown in SEQ ID NO: 1341 or a portion thereof. In some embodiments, the antisense oligonucleotide is complementary to the sequence shown in SEQ ID NO: 1342 or a portion thereof. In some embodiments, the antisense oligonucleotide is complementary to at least 1, at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, at least 11, at least 12, or at least 13 nucleotides of the sequence shown in any one of SEQ ID NOs: 1339-1342. In some embodiments, the antisense oligonucleotide is complementary to at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, at least 11, at least 12, or at least 13 consecutive nucleotides of the sequence shown in any one of SEQ ID NOs: 1339-1342. In some embodiments, the antisense oligonucleotide is at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99%, or 100% complementary to the sequence shown in SEQ ID NO: 1339. In some embodiments, the antisense oligonucleotide is at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99%, or 100% complementary to the sequence shown in SEQ ID NO: 1340. In some embodiments, the antisense oligonucleotide is at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99%, or 100% complementary to the sequence shown in SEQ ID NO: 1341. In some embodiments, the antisense oligonucleotide is at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99%, or 100% complementary to the sequence shown in SEQ ID NO: 1342.

[0032] The present invention also provides an antisense oligonucleotide, which comprises the sequence shown in any one of SEQ ID NO: 20-355, 400-638, 810-959 or 961-1332. In some embodiments, the antisense oligonucleotide consists of 13-30 nucleotides. In some embodiments, the antisense oligonucleotide consists of 13-25 nucleotides. In some embodiments, the antisense oligonucleotide consists of 13, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24 or 25 nucleotides. In some embodiments, the antisense oligonucleotide consists of 19, 20, 21 or 22 nucleotides. In some embodiments, the antisense oligonucleotide is complementary to the sequence shown in SEQ ID NO: 1.

[0033] The present invention also provides an antisense oligonucleotide, which consists of the sequence shown in any one of SEQ ID NO: 20-355, 400-638, 810-959 or 961-1332. The present invention also provides an antisense oligonucleotide, wherein the antisense oligonucleotide is complementary to the sequence shown in any one of SEQ ID NO: 1339-1342 or a part thereof.

[0034] In some embodiments, the antisense oligonucleotide consists of 13 nucleotides complementary to the sequence shown in SEQ ID NO:1, and wherein the antisense oligonucleotide consists of the sequence shown in any one of SEQ ID NOs: 961-983 and 1116-1145. In some embodiments, the antisense oligonucleotide consists of 15 nucleotides complementary to the sequence shown in SEQ ID NO:1, and wherein the antisense oligonucleotide consists of the sequence shown in any one of SEQ ID NOs: 984-1004 and 1146-1173. In some embodiments, the antisense oligonucleotide consists of 16 nucleotides complementary to the sequence shown in SEQ ID NO:1, and wherein the antisense oligonucleotide consists of the sequence shown in any one of SEQ ID NOs: 41, 42, 56, 57, 71, 72, 86, 87, 101, 102, 116, 117, 131, 132, 146, 147, 161, 162, 176, 177, 191, 192, 206, 207, 221, 222, 236, 237, 251, 252, 266, 267, 281, 282, 296, 297, 311, 326, 341, 810, 811, 812, 813, 826, 827, 835, 838, 841, 842, 851, 852, 861, 864, 867, 868, 877, 878, 895, 900, 901, 910, 934 and 935. In some embodiments, the antisense oligonucleotide consists of 17 nucleotides complementary to the sequence shown in SEQ ID NO:1, and wherein the antisense oligonucleotide consists of the sequence shown in any one of SEQ ID NOs: 43, 58, 73, 88, 103, 118, 133, 148, 163, 178, 193, 208, 223, 238, 253, 268, 283, 298, 312, 313, 327, 328, 342, 343, 814, 815, 816, 828, 829, 836, 839, 843, 844, 853, 854, 862, 865, 869, 870, 879, 880, 881, 891, 896, 897, 902, 903, 1005-1023 and 1174-1199.In some embodiments, the antisense oligonucleotide consists of 18 nucleotides complementary to the sequence shown in SEQ ID NO:1, and wherein the antisense oligonucleotide consists of the sequence shown in any one of SEQ ID NOs: 38-40, 44, 45, 59, 60, 74, 75, 89, 90, 104, 105, 119, 120, 134, 135, 149, 150, 164, 165, 179, 180, 194, 195, 209, 210, 224, 225, 239, 240, 254, 255, 269, 270, 284, 285, 299, 300, 314, 329, 344, 518-638, 817, 818, 845, 855, 871, 882, 904, 958, 959, 960, 1024-1041 and 1200-1224. In some embodiments, the antisense oligonucleotide consists of 19 nucleotides complementary to the sequence shown in SEQ ID NO:1, and wherein the antisense oligonucleotide consists of the sequence shown in any one of SEQ ID NOs: 46, 61, 76, 91, 106, 121, 136, 151, 166, 181, 196, 211, 226, 241, 256, 271, 286, 301, 315, 316, 330, 331, 345, 346, 819, 830, 846, 856, 872, 883, 884, 892, 905, 1042-1058 and 1225-1247. In some embodiments, the antisense oligonucleotide consists of 20 nucleotides complementary to the sequence shown in SEQ ID NO:1, and wherein the antisense oligonucleotide consists of the sequence shown in any one of SEQ ID NOs: 47, 48, 62, 63, 77, 78, 92, 93, 107, 108, 122, 123, 137, 138, 152, 153, 167, 168, 182, 183, 197, 198, 212, 213, 227, 228, 242, 243, 257, 258, 272, 273, 287, 288, 302, 303, 317, 332, 347, 911, 936, 937, 1059-1074 and 1248-1270.In some embodiments, the antisense oligonucleotide consists of 21 nucleotides complementary to the sequence shown in SEQ ID NO:1, and wherein the antisense oligonucleotide consists of the sequence shown in any one of SEQ ID NOs: 20-37, 49, 64, 79, 94, 109, 124, 139, 154, 169, 184, 199, 214, 229, 244, 259, 274, 289, 304, 318, 319, 333, 334, 348, 349, 400-517, 1075-1089, and 1271-1292. In some embodiments, the antisense oligonucleotide consists of 22 nucleotides complementary to the sequence shown in SEQ ID NO:1, and wherein the antisense oligonucleotide consists of the sequence shown in any one of SEQ ID NOs: 50, 51, 65, 66, 80, 81, 95, 96, 110, 111, 125, 126, 140, 141, 155, 156, 170, 171, 185, 186, 200, 201, 215, 216, 230, 231, 245, 246, 260, 261, 275, 276, 290, 291, 305, 306, 320, 335, 350, 912, 913, 914, 938, 939, 940, 941, 1090-1103, and 1293-1313. In some embodiments, the antisense oligonucleotide consists of 23 nucleotides complementary to the sequence shown in SEQ ID NO:1, and wherein the antisense oligonucleotide consists of the sequence shown in any one of SEQ ID NOs: 52, 67, 82, 97, 112, 127, 142, 157, 172, 187, 202, 217, 232, 247, 262, 277, 292, 307, 321, 322, 336, 337, 351, 352, 820, 831, 847, 857, 873, 885, 886, 893, 906, 915-918, 930, and 942-945.In some embodiments, the antisense oligonucleotide consists of 24 nucleotides complementary to the sequence shown in SEQ ID NO:1, and wherein the antisense oligonucleotide consists of the sequence shown in any one of SEQ ID NO:53, 54, 68, 69, 83, 84, 98, 99, 113, 114, 128, 129, 143, 144, 158, 159, 173, 174, 188, 189, 203, 204, 218, 219, 233, 234, 248, 249, 263, 264, 278, 279, 293, 294, 308, 309, 323, 338, 353, 821, 822, 832, 848, 858, 874, 887, 907, 919, 920 - 923, 931, 946 - 951, 1104 - 1115 and 1314 - 1332. In some embodiments, the antisense oligonucleotide consists of 25 nucleotides complementary to the sequence shown in SEQ ID NO:1, and wherein the antisense oligonucleotide consists of the sequence shown in any one of SEQ ID No:55, 70, 85, 100, 115, 130, 145, 160, 175, 190, 205, 220, 235, 250, 265, 280, 295, 310, 324, 325, 339, 340, 354, 355, 823, 824, 825, 833, 834, 837, 840, 849, 850, 859, 860, 863, 866, 875, 876, 888 - 890, 894, 898, 899, 908, 909, 924 - 929, 932, 933, 952 and 953 - 957.

[0035] In some embodiments, the antisense oligonucleotide is complementary to the sequence shown in SEQ ID NO:1339 or a portion thereof. In some embodiments, the antisense oligonucleotide is complementary to the sequence shown in SEQ ID NO:1340 or a portion thereof. In some embodiments, the antisense oligonucleotide is complementary to the sequence shown in SEQ ID NO:1341 or a portion thereof. In some embodiments, the antisense oligonucleotide is complementary to the sequence shown in SEQ ID NO:1342 or a portion thereof. In some embodiments, the antisense oligonucleotide is complementary to at least 1, at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, at least 11, at least 12 or at least 13 nucleotides of the sequence shown in any one of SEQ ID NO:1339 - 1342.

[0036] In some embodiments, the antisense oligonucleotide is complementary to at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, at least 11, at least 12, or at least 13 consecutive nucleotides of the sequence shown in any one of SEQ ID NOs: 1339 - 1342. In some embodiments, the antisense oligonucleotide is at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99%, or 100% complementary to the sequence shown in SEQ ID NO: 1339. In some embodiments, the antisense oligonucleotide is at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99%, or 100% complementary to the sequence shown in SEQ ID NO: 1340. In some embodiments, the antisense oligonucleotide is at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99%, or 100% complementary to the sequence shown in SEQ ID NO: 1341. In some embodiments, the antisense oligonucleotide is at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99%, or 100% complementary to the sequence shown in SEQ ID NO: 1342. In some embodiments, the antisense oligonucleotide consists of the sequence shown in any one of SEQ ID NOs: 1059 - 1074, 1075 - 1089, 1090 - 1103, 1248 - 1270, 1271 - 1292, and 1293 - 1313.

[0037] In some embodiments, the antisense oligonucleotide comprises a modification. In some embodiments, the 5'-end and / or 3'-end of the antisense oligonucleotide comprises a modification. In some embodiments, the 5'-end and 3'-end of the antisense oligonucleotide comprise modifications, wherein 2, 3, 4, or 5 nucleotides are modified at each of the 5'-end and 3'-end of the antisense oligonucleotide. In some embodiments, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 95%, or at least 98% of the nucleotides comprise a modification. In some embodiments, the modification is selected from the group consisting of: 2'-OMe antisense oligonucleotide, 2'-O-methoxyethyl antisense oligonucleotide, phosphorothioate antisense oligonucleotide, and locked nucleic acid (LNA) antisense oligonucleotide. In some embodiments, the modification is a 2'-OMe antisense oligonucleotide.

[0038] Also provided herein is a pharmaceutical composition, the pharmaceutical composition comprising the antisense oligonucleotide of any one of the embodiments disclosed herein. In some embodiments, the pharmaceutical composition comprises an excipient and / or a buffer.

[0039] The present invention also provides a kit, which comprises the antisense oligonucleotides of some embodiments disclosed herein or the pharmaceutical compositions of some embodiments disclosed herein.

[0040] All publications, including patent documents, scientific articles, and databases, mentioned in this application are incorporated herein by reference in their entirety for all purposes to the extent as if each individual publication were incorporated by reference separately. If the definitions set forth herein are contrary to or otherwise inconsistent with the definitions set forth in the patents, applications, published applications, and other publications incorporated herein by reference, the definitions set forth herein shall prevail over the definitions incorporated herein by reference.

[0041] The section headings used herein are for organizational purposes only and should not be construed as limiting the subject matter described. BRIEF DESCRIPTION OF THE DRAWINGS

[0042] The drawings illustrate certain features and advantages of the present disclosure. These embodiments are not intended to limit the scope of the appended claims in any way.

[0043] Figure 1A and Figure 1B Shows the percentage of UNC13A mRNA after treatment with ASO in TDP-43 knockdown iPSC-derived motor neurons compared to treatment with a control ASO that does not affect UNC13A mRNA. UNC13A mRNA was detected using qPCR. The position of the ASO target is plotted from 5' to 3'.

[0044] Figure 2A and Figure 2B Shows the percentage of UNC13A mRNA containing a cryptic exon (CE) after treatment with ASO in TDP-43 knockdown iPSC-derived motor neurons compared to treatment with a control ASO that does not affect UNC13A CE mRNA. UNC13A CE mRNA was detected using qPCR. The position of the ASO target is plotted from 5' to 3'.

[0045] Figure 3A and Figure 3B Shows the percentage of UNC13A mRNA, the percentage of UNC13A mRNA containing a cryptic exon (CE), and the expression of a control transcript STMN2 that is not targeted by the ASO in TDP-43 knockdown iPSC-derived motor neurons. The position of the ASO target is plotted from 5' to 3'.

[0046] Figure 4Shows the percentage of UNC13A cryptic exon transcripts and the percentage of UNC13A full-length transcripts for each of the ASOs tested in TDP-43 knockdown iPSC-derived motor neurons. ASOs that reduce UNC13A CE and increase UNC13A full-length transcripts are shown as squares.

[0047] Figure 5A and Figure 5B is a map of the UNC13A cryptic exon, showing the positions of the active oligonucleotides shown in dark gray and the inert oligonucleotides shown in light gray. Figure 5B Shows four identified regulatory regions.

[0048] Figure 6A and Figure 6B Shows the effect of ASOs on UNC13A expression in TDP43 knockdown iPSC-derived motor neurons. NT, non-targeting ASO; ASO 434, inert ASO; +NT, non-targeting shRNA; +shTDP43, shRNA to TARDBP. Figure 6C Shows the effect of ASOs on TARDBP expression in iPSC-derived motor neurons treated with shRNA to TDP-43 compared to control cells treated with an inactive ASO. Figure 6D Shows the effect of ASOs on STMN2-expressing iPSC-derived motor neurons treated with shRNA to TDP-43 compared to control cells treated with an inactive ASO. NT, non-targeting ASO; ASO 434, inert ASO; +NT, non-targeting shRNA; +shTDP43, shRNA to TARDBP.

[0049] Figure 7A Shows the effect of ASO 422 on the percentage of UNC13A mRNA expression in TDP-43 knockdown iPSC-derived motor neurons in extended culture compared to no ASO or a control ASO that does not target UNC13A. Figure 7B Shows the effect of ASO 422 on the UNC13A cryptic exon (CE) in TDP-43 knockdown iPSC-derived motor neurons in extended culture compared to no ASO or a control ASO that does not target UNC13A. Figure 7C Shows the effect of ASO 422 on TARDBP expression in TDP-43 knockdown iPSC-derived motor neurons in extended culture compared to no ASO or a control ASO that does not target UNC13A.

[0050] Figure 8AShows a Western blot of UNC13 protein levels in TDP-43 knockdown iPSC-derived motor neurons treated with ASO or a control ASO (432) that does not target UNC13A. Figure 8B Shows the quantification of UNC13A protein levels determined by Western blot.

[0051] Figure 9A and Figure 9B Shows the percentage of UNC13A mRNA expression after treatment with different doses of ASO (0.024 - 3 μM) compared to a control ASO that does not affect UNC13A mRNA expression. Figure 9C and Figure 9D Shows the percentage of UNC13A mRNA containing a cryptic exon (CE) after treatment with different doses of ASO (0.024 - 3 μM) compared to a control ASO that does not affect UNC13A mRNA expression. Figure 9E and Figure 9F Shows the expression levels of TARDBP and STMN2 after treatment with different doses of ASO. After ASO delivery, cells were transduced with shRNA against TARDBP or a control. qPCR was used to detect the mRNA expression levels of UNC13A, UNC13A CE, STMN2, and TARDBP. ASO 432, inert ASO; +NT, non-targeting shRNA; +shTDP43, shRNA to TARDBP.

[0052] Figure 10A Shows the percentage of UNC13A protein expression in iPSC-derived motor neurons with TDP-43 knockdown treated with different concentrations (0.024 - 3 μM) of ASO alone. Figure 10B Shows the percentage of UNC13A protein expression under all ASO treatment conditions. Figure 10C Shows the rank order of HiBiT signals (UNC13A protein expression) in neurons treated with 3 μM ASO. Figure 10D Shows the percentage of UNC13A mRNA and UNC13A protein expression for each control and UNC13A splicing-correcting ASO tested.

[0053] Figure 11A Shows the percentage of UNC13A mRNA expression after treatment with ASO (0.024 μM or 3 μM) compared to a control ASO that does not affect UNC13A mRNA expression in iPSC-derived motor neurons with TDP-43 knockdown. Figure 11BShows the percentage of UNC13A protein expression after treatment with ASO (0.024 μM or 3 μM) compared to a control ASO that does not affect UNC13A mRNA expression. Figure 11C Shows the percentage of UNC13A mRNA and UNC13A protein expression for each control and UNC13A splicing-correcting ASO tested. After ASO delivery, cells were transduced with shRNA against TARDBP or a control. UNC13A, UNC13A CE, STMN2, and TARDBP mRNA expression levels were detected using qPCR. ASO 432, inert ASO; +NT, non-targeting shRNA; +shTDP43, shRNA to TARDBP.

[0054] Figure 12A and Figure 12B Shows the percentage of UNC13A and UNC13A CE mRNA expression after treatment with ASO (0.024 μM or 3 μM) compared to a control ASO that does not affect UNC13A mRNA expression in GM25256 (GM) iPSC-derived motor neurons. After ASO delivery, cells were transduced with shRNA against TARDBP or a control.

[0055] Figure 13A and Figure 13B Shows the percentage of UNC13A and UNC13A CE mRNA expression after treatment with ASO (0.024 μM or 3 μM) compared to a control ASO that does not affect UNC13A mRNA expression in NCRM-5 (005) iPSC-derived motor neurons. After ASO delivery, cells were transduced with shRNA against TARDBP or a control.

[0056] Figure 14A and Figure 14B Shows the percentage of UNC13A and UNC13A CE mRNA expression after treatment with ASO (0.024 μM or 3 μM) compared to a control ASO that does not affect UNC13A mRNA expression in NDS00209 (4048) iPSC-derived motor neurons. After ASO delivery, cells were transduced with shRNA against TARDBP or a control.

[0057] Figure 15A and Figure 15BShows the percentage of UNC13A and UNC13A CE mRNA expression after treatment with ASO (0.024 μM or 3 μM) compared to a control ASO that does not affect UNC13A mRNA expression in NDS00235(4157) iPSC-derived motor neurons. After ASO delivery, cells were transduced with shRNA against TARDBP or a control.

[0058] Figure 16A and Figure 16B Shows cumulative UNC13A and UNC13A CE mRNA expression data from four iPSC cell lines (GM, 005, 4048, and 4157). The percentage of UNC13A and UNC13A CE mRNA expression after treatment with UNC13A splicing-blocking ASO (0.024 μM or 3 μM) was plotted. After ASO delivery, cells were transduced with shRNA against TARDBP or a control.

[0059] Figure 17A - 17H Shows the percentage of UNC13A mRNA expression ([ Figure 17A and Figure 17B ) after extended culture and multiple doses of UNC13A splicing-correcting ASO or control non-targeting ASO (432) in GM and 005 iPSC-derived motor neurons Figure 17C and Figure 17D ), the percentage of UNC13A CE mRNA expression ([ Figure 17E and Figure 17G ), the percentage of TARDBP mRNA expression ([ Figure 17F and Figure 17H ), and the percentage of STMN2 mRNA expression ([

[0060] Figure 18A ). Before administration of the UNC13A splicing-correcting ASO, GM and 005 cells were treated with a gapmer ASO against TARDBP or a non-targeting control gapmer. Figure 18B Shows the percentage of nuclear area of neurons treated with UNC13A splicing-blocking ASO.

[0061] Figure 19A - 19R Shows the percentage of UNC13A and UNC13A CE mRNA expression after treatment with different lengths of ASO (3 μM) targeting region 2 of UNC13A CE. Figure 19A (13-mer), Figure 19C (15-mer), Figure 19E (17-mer), Figure 19G (18-mer), Figure 19I (19-mer),Figure 19K (20-mer), Figure 19M (21-mer), Figure 19O (22-mer) and Figure 19Q (24-mer) show the percentage of UNC13A mRNA expression. Figure 19B (13-mer), Figure 19D (15-mer), Figure 19F (17-mer), Figure 19H (18-mer), Figure 19J (19-mer), Figure 19L (20-mer), Figure 19N (21-mer), Figure 19P (22-mer) and Figure 19R (24-mer) show the percentage of UNC13A CE mRNA expression. After ASO delivery, iPSC-derived motor neurons were transduced with shRNA against TARDBP or a control.

[0062] Figure 20A - 20R Show the percentage of UNC13A and UNC13A CE mRNA expression after treatment with different lengths of ASO (3 μM) targeting region 4 of UNC13A CE. Figure 20A (13-mer), Figure 20C (15-mer), Figure 20E (17-mer), Figure 20G (18-mer), Figure 20I (19-mer), Figure 20K (20-mer), Figure 20M (21-mer), Figure 20O (22-mer) and Figure 20Q (24-mer) show the percentage of UNC13A mRNA expression. Figure 20B (13-mer), Figure 20D (15-mer), Figure 20F (17-mer), Figure 20H (18-mer), Figure 20J (19-mer), Figure 20L (20-mer), Figure 20N (21-mer), Figure 20P (22-mer) and Figure 20R (24-mer) show the percentage of UNC13A CE mRNA expression. After ASO delivery, iPSC-derived motor neurons were transduced with shRNA against TARDBP or a control.

[0063] Figure 21A Show the average percentage of UNC13A mRNA expression from all cells treated with sub-ASOs of the specified length contained within the left sequence. The darker shading indicates higher UNC13A mRNA expression.Figure 21B Shows the average difference in the percentage of UNC13A mRNA expression between cells treated with the parental 24-mer ASO shown on the left and all cells treated with the sub-ASOs of the indicated lengths contained within the parental sequence. Lighter shading indicates an increase in UNC13A mRNA expression for the sub-ASO of that length compared to the parental 24-mer, while darker shading indicates a decrease in UNC13A mRNA expression for the sub-ASO of that length compared to the parental 24-mer. After ASO delivery, cells were transduced with shRNA against TARDBP or a control.

[0064] Figure 22A - 22B Shows the effect of ASOs in the range of 13 nucleotides (13-mer) to 24 nucleotides (24-mer) on UNC13A mRNA expression in cells transduced with shRNA against TARDBP after ASO delivery, i.e., region 2 ( Figure 22A ) and region 4 ( Figure 22B ). The top part of the figure plots the average percentage of UNC13A expression for all ASOs grouped by ASO length that overlap each nucleotide indicated. Detailed Description

[0065] TDP-43 depletion is associated with multiple neuronal pathologies including ALS and FTD. Depletion of TDP-43 results in incorrect splicing of UNC13A mRNA and inclusion of a cryptic exon between exons 20-21. Inclusion of the UNC13A cryptic exon results in reduced UNC13A protein and is associated with such neuronal pathologies. Antisense oligonucleotides targeting UNC13 mRNA are provided herein that reduce inclusion of the UNC13A cryptic exon by targeting four regions of the UNC13A mRNA transcript: the 5' splice donor site, the 3' splice acceptor site, and two regions within the cryptic exon itself. The antisense oligonucleotides are capable of restoring UNC13A expression in neurons and increasing the ratio of correctly spliced UNC13A transcripts to incorrectly spliced UNC13A transcripts.

[0066] Definitions

[0067] Before elaborating on the present disclosure in more detail, providing definitions of certain terms used herein may aid in understanding the present disclosure. Additional definitions are set forth throughout the present disclosure.

[0068] In this specification, unless otherwise indicated, any concentration range, percentage range, ratio range, or integer range should be understood to include any integer value within the range and, where appropriate, its fractions (such as tenths and hundredths of an integer) or sub-ranges. The description of a range includes the description of each endpoint of the range.

[0069] As used herein, unless otherwise indicated, the term "about" means ±20% of the indicated range, value, or structure.

[0070] It should be understood that, as used herein, the terms "a" and "an" refer to "one or more" of the recited components. The use of alternative (e.g., "or") should be understood to mean one, both, or any combination of the alternatives.

[0071] As used herein, the terms "include", "have", and "comprise" are used synonymously, and these terms and their variants are intended to be construed as non-limiting.

[0072] "Optional" or "optionally" means that the subsequently described element, component, event, or circumstance may or may not occur, and the description includes the case where the element, component, event, or circumstance occurs and the case where the element, component, event, or circumstance does not occur.

[0073] As used herein, "nucleic acid" or "nucleic acid molecule" or "polynucleotide" refers to any deoxyribonucleic acid (DNA), ribonucleic acid (RNA), oligonucleotide, molecules produced, for example, by polymerase chain reaction (PCR) or by in vitro translation, and molecules produced by ligation, cleavage, endonuclease action, exonuclease action, or mechanical action (e.g., shearing). Nucleic acids can be composed of a variety of monomers, which are naturally occurring nucleotides (such as deoxyribonucleotides and ribonucleotides), analogs of naturally occurring nucleotides (e.g., α - enantiomeric forms of naturally occurring nucleotides), or a combination of both. Modified nucleotides can have modifications or substitutions of the sugar moiety or the pyrimidine or purine base moiety (e.g., morpholino nucleotides). The nucleic acid monomers of a polynucleotide can be linked by phosphodiester bonds or analogs of such bonds. Analogs of phosphodiester bonds include phosphorothioates, dithiophosphates, selenophosphates, diselenophosphates, phosphorothioanilates, anilophosphates, aminophosphates, etc. Nucleic acid molecules can be single-stranded or double-stranded.

[0074] As used herein, "protein" or "polypeptide" refers to a compound composed of amino acid residues covalently linked by peptide bonds. The term "protein" may be synonymous with the term "polypeptide" or alternatively may refer to a complex of two or more polypeptides. In certain embodiments, the polypeptide may be a fragment. As used herein, "fragment" means a polypeptide lacking one or more amino acids found in a reference sequence. A fragment may contain a binding domain, antigen, or epitope found in the reference sequence. A fragment of a polypeptide may have at least about 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more of the amino acids of the amino acid sequence of the reference sequence.

[0075] The term "isolated" means that a material, complex, compound, or molecule has been removed from its original environment (e.g., the natural environment if the original environment is a naturally occurring one). For example, a naturally occurring polynucleotide or polypeptide present in a living animal is not isolated, but the same polynucleotide or polypeptide separated from some or all of the coexisting materials in the natural system is isolated. Such nucleic acids may be part of a vector and / or such nucleic acids or polypeptides may be part of a composition (e.g., a cell lysate) and still be isolated because such a vector or composition is not part of the natural environment of the nucleic acid or polypeptide. The term "gene" means a DNA segment involved in the production of a polypeptide chain; it includes the regions before and after the coding region "leader and tail" and the intervening sequences (introns) between individual coding segments (exons) (if any).

[0076] As used herein, the terms "recombinant" or "genetically engineered" refer to a cell, microorganism, nucleic acid molecule, polypeptide, or vector that has been genetically modified by human intervention. For example, a recombinant polynucleotide is modified by the introduction of an exogenous or heterologous nucleic acid molecule by a human or a machine, or refers to a cell or microorganism that has been altered by human or machine intervention such that the expression of an endogenous nucleic acid molecule or gene is controlled, deregulated, or constitutive. Genetically engineered changes made by a human may include, for example, modifications that introduce a nucleic acid molecule encoding one or more proteins or enzymes (which may include expression control elements such as promoters), or the addition, deletion, substitution, or other functional disruption or addition to the genetic material of the cell or the encoded product. Exemplary modifications introduced by a human or a machine include those in the coding region or a functional fragment thereof of a heterologous or homologous polypeptide from a reference or parental molecule.

[0077] A "wild-type" gene or gene product is the gene or gene product that is most frequently observed in a population and is thus arbitrarily designated as the "normal" or "reference" or "wild-type" form of the gene.

[0078] As used herein, a "mutation" refers to a change in the sequence of a nucleic acid molecule or polypeptide molecule as compared to a reference or wild-type nucleic acid molecule or polypeptide molecule, respectively. A mutation can give rise to several different types of changes in the sequence, including nucleotide or amino acid substitutions, insertions, or deletions.

[0079] A "conservative substitution" is an amino acid substitution that does not significantly affect or alter the binding properties of a particular protein. Generally, a conservative substitution is a substitution in which the amino acid residue being replaced is replaced with an amino acid residue having a similar side chain. Conservative substitutions include substitutions found in one of the following groups: Group 1: alanine (Ala or A), glycine (Gly or G), serine (Ser or S), threonine (Thr or T); Group 2: aspartic acid (Asp or D), glutamic acid (Glu or Z); Group 3: asparagine (Asn or N), glutamine (Gln or Q); Group 4: arginine (Arg or R), lysine (Lys or K), histidine (His or H); Group 5: isoleucine (Ile or I), leucine (Leu or L), methionine (Met or M), valine (Val or V); and Group 6: phenylalanine (Phe or F), tyrosine (Tyr or Y), tryptophan (Trp or W). Additionally or alternatively, amino acids can be grouped into conservative substitution groups based on similar function, chemical structure, or composition (e.g., acidic, basic, aliphatic, aromatic, or sulfur-containing). For example, for the purposes of substitution, the aliphatic grouping can include Gly, Ala, Val, Leu, and Ile. Other conservative substitution groups include: sulfur-containing: Met and cysteine (Cys or C); acidic: Asp, Glu, Asn, and Gln; small aliphatic, nonpolar, or slightly polar residues: Ala, Ser, Thr, Pro, and Gly; polar negatively charged residues and their amides: Asp, Asn, Glu, and Gln; polar positively charged residues: His, Arg, and Lys; large aliphatic, nonpolar residues: Met, Leu, Ile, Val, and Cys; and large aromatic residues: Phe, Tyr, and Trp. Additional information can be found in Creighton (1984) Proteins, W.H. Freeman and Company.

[0080] As used herein, the term "expression" refers to the process of producing a polypeptide based on the coding sequence of a nucleic acid molecule such as a gene. The process can include transcription, post-transcriptional control, post-transcriptional modification, translation, post-translational control, post-translational modification, or any combination thereof.

[0081] As used herein, "sequence identity" refers to the percentage of nucleotides (amino acid residues) in a sequence that are identical to the nucleotides (amino acid residues) in another reference polynucleotide (polypeptide) sequence after aligning the sequences and introducing gaps, if necessary, to achieve the maximum percentage sequence identity and not considering any conservative substitutions as part of the sequence identity. The sequence identity percentage value can be generated using the NCBI BLAST 2.0 software as defined in Altschul et al. (1997) "Gapped BLAST and PSI-BLAST: a new generation of protein database search programs", Nucleic Acids Res. 25: 3389-3402, where the parameters are set to the default values.

[0082] As used herein, "UNC13A" refers to a presynaptic protein that regulates the release of neurotransmitters, peptides, and hormones and is found in central and neuromuscular synapses. The UNC13A reference or wild-type mRNA transcript contains 44 exons encoding a 1,703 amino acid protein. In an embodiment, the NCBI reference sequence: NP_001073890.2 (SEQ ID NO: 11) is an example of a wild-type or reference UNC13A protein. In an embodiment, the NCBI reference sequence NM_001080421.3 (SEQ ID NO: 1) is an example of a wild-type or reference UNC13A mRNA transcript. In an embodiment, UNC13A includes all forms of UNC13A, including wild-type, spliced isoforms, variants, mutants, native conformations, misfolded forms, and post-translational modifications. In an embodiment, UNC13A does not include UNC13A cryptic exon splicing variants.

[0083] As used herein, the term "preprocessed mRNA" or "precursor mRNA" or "precursor mRNA" refers to the primary transcript synthesized from a DNA template and that has not yet undergone processing (e.g., splicing, addition of a 5' cap, and addition of a 3' polyA tail) to become mature mRNA. Mature mRNA can be translated into protein by ribosomes.

[0084] As used herein, the term "cryptic exon" or "pseudoexon" refers to an exon that is not present or undetectably used in wild-type precursor mRNA but is selected in variant isoforms. Cryptic exons can be generated possibly due to mutations that create new splice sites or remove existing binding sites for splicing repressors. Cryptic exons can also arise from transposable elements (e.g., Alu elements).

[0085] As used herein, "UNC13A cryptic exon splicing variant" refers to an mRNA that contains a cryptic exon between exon 20 and exon 21 or a protein encoded by said mRNA. The cryptic exon is derived from intron 20-21 of the UNC13A gene. In an embodiment, the cryptic exon has the nucleotide sequence of SEQ ID NO:5 or SEQ ID NO:6. In an embodiment, the UNC13A cryptic exon splicing variant can have the nucleotide sequence of SEQ ID NO:7 encoding the protein sequence of SEQ ID NO:8 or the nucleotide sequence of SEQ ID NO:9 encoding the protein sequence of SEQ ID NO:10.

[0086] As used herein, "trans-activation response element DNA-binding protein 43" or "TAR-DNA-binding protein-43" or "TDP-43" refers to a protein typically having 414 amino acid residues encoded by TARDBP as well as the mRNA transcript of TARDBP. In some cases, the mRNA encoded by the TARDBP gene is referred to herein as TDP-43. In an embodiment, the wild-type TDP-43 amino acid sequence is provided by Uniprot accession number Q13148 (SEQ ID NO:378). In an embodiment, TDP-43 includes all forms of TDP-43, including proteins in wild-type, spliced isoforms, variants, mutants, native conformations, misfolded, and post-translational modifications (e.g., ubiquitination, phosphorylation, acetylation, sumoylation, or cleavage into C-terminal fragments).

[0087] As used herein, "TAR-DNA-binding protein-43 proteinopathy" or "TDP-43 proteinopathy" refers to a neurodegenerative disease characterized by the deposition of TDP-43 positive protein inclusions in the brain and / or spinal cord of a subject. Hyperphosphorylation, ubiquitination, and cytoplasmic inclusions in cleaved forms of TDP-43 are pathological features of the disease, including but not limited to amyotrophic lateral sclerosis (ALS), frontotemporal lobar degeneration (FTLD), primary lateral sclerosis (PLS), progressive muscular atrophy (PMA), facial onset sensory and motor neuronopathy (FOSMN), hippocampal sclerosis (HS), limbic-predominant age-related TDP-43 encephalopathy (LATE), cerebrum age-related TDP-43 with sclerosis (CARTS), Guam Parkinson-dementia complex (G-PDC), GuanALS (G-ALS), multisystem proteinopathy (MSP), Perry disease, Alzheimer's disease (AD), and chronic traumatic encephalopathy (CTE).

[0088] The terms "complementary" and "complementarity" refer to polynucleotides (i.e., nucleotide sequences) related by base-pairing rules. For example, the sequence "A-G-T" is complementary to the sequence "T-C-A". Complementarity can be "partial", where only some of the bases of the nucleic acids match according to the base-pairing rules, or there can be "perfect" or "total" complementarity between the nucleic acids. The degree of complementarity between nucleic acid strands has a significant impact on the efficiency and strength of hybridization between the nucleic acid strands. Although perfect complementarity is generally desired, some embodiments can include one or more but preferably 6, 5, 4, 3, 2, or 1 mismatch relative to a target nucleic acid (e.g., RNA). Variations at any position within the oligomer are included. In certain embodiments, sequence variations near the ends of the oligomer are generally preferred over internal variations and typically within about 6, 5, 4, 3, 2, or 1 nucleotides at the 5' and / or 3' ends if present.

[0089] The terms "antisense oligomer" or "antisense compound" or "antisense oligonucleotide" or "oligonucleotide" are used interchangeably and refer to short single-stranded polynucleotides (e.g., 10 - 50 subunits) composed of DNA, RNA, or both, which hybridize to a target sequence in a nucleic acid (typically RNA) via Watson-Crick base pairing to form a nucleic acid:oligomer heteroduplex within the target sequence. Antisense oligonucleotides can contain unmodified nucleotides or can contain modified nucleotides, non-natural nucleotides, or nucleotide analogs such as morpholino, phosphorothioate, peptide nucleic acid, LNA, 2'-O-Me RNA, 2'F-RNA, 2'-O-MOE-RNA, 2'F-ANA, or any combination thereof.

[0090] Such antisense oligomers can be designed to block or inhibit translation of mRNA or inhibit natural pre-mRNA splicing processing, or induce degradation of the targeted mRNA, and can be referred to as "directed against" or "targeting" the target sequence to which they hybridize. In embodiments, the target sequence is a region surrounding or including the AUG start codon of mRNA, the 3' or 5' splice site of pre-processed mRNA, or the branch point. The target sequence can be within an exon or within an intron or a combination thereof. The target sequence of a splice site can include an mRNA sequence that has its 5' end 1 to about 25 base pairs downstream of the normal splice acceptor junction in pre-processed mRNA. Exemplary target sequences of splice sites are any regions of pre-processed mRNA that include the splice site or are completely contained within an exon coding sequence or span the splice acceptor or donor site. When an oligomer targets a nucleic acid of a target in the above manner, the oligomer is more generally said to be "targeted" to a biologically relevant target, such as the human UNC13A gene pre-mRNA encoding the UNC13A protein in the present disclosure. Exemplary target sequences include the target sequences listed in Tables 2 - 5.

[0091] The term "oligonucleotide analog" refers to an oligonucleotide having: (i) a modified backbone structure, e.g., a backbone other than the standard phosphodiester bond found in natural oligonucleotides and polynucleotides, and (ii) optionally, a modified sugar moiety, e.g., a morpholino moiety rather than a ribose or deoxyribose moiety. The oligonucleotide analog supports bases capable of hydrogen bonding with standard polynucleotide bases via Watson-Crick base pairing, wherein the analog backbone presents the bases in a manner that permits such hydrogen bonding to occur in a sequence-specific manner between the bases in the oligonucleotide analog molecule and a standard polynucleotide (e.g., single-stranded RNA or single-stranded DNA). Exemplary analogs are those having a substantially uncharged phosphorus-containing backbone.

[0092] As used herein, "modified nucleotide" is a nucleotide other than a ribonucleotide (2'-hydroxynucleotide). In some embodiments, an antisense oligonucleotide (ASO) contains one or more modified nucleotides. In some embodiments, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 95% or 100% of the nucleotides are modified. Modified nucleotides include, but are not limited to, deoxynucleotides, nucleotide analogs, abasic nucleotides (represented herein as x or ab), 2'-modified nucleotides, 3' to 3' bond (inverted) nucleotides (represented herein as invdn, invn, invn, invx), non-natural bases containing nucleotides, bridged nucleotides, peptide nucleic acids, 2',3'-cleaved nucleotide analogs (unlocked nucleobase analogs, represented herein as nuna or nuna), locked nucleotides (represented herein as nlna or nlna), 3'-O-methoxy (2'-nucleotide internucleotide linked) nucleotides (represented herein as 3'-omen), 2'-F-arabinonucleotides (represented herein as nfana or nfana), morpholino nucleotides, vinyl phosphonate deoxyribonucleotides (represented herein as vpdn) and vinyl phosphonate nucleotides (represented herein as vpn). 2'-modified nucleotides (i.e., nucleotides having a group other than a hydroxyl group at the 2' position of the pentose sugar ring) include, but are not limited to, 2'-methyl nucleotides (represented as the lowercase letter "n" in a nucleotide sequence), 2'-deoxy-2'-fluoronucleotides (represented herein as nf, also represented herein as 2'-fluoronucleotide), 2'-deoxynucleotides (represented herein as dn), 2'-methoxyethyl (2'-O-2-methoxyethyl) nucleotides (represented herein as nm or 2'-moe), 2'-aminonucleotides and 2'-alkyl nucleotides. All positions in a given compound need not be uniformly modified. Instead, more than one modification can be incorporated into a single ASO or even into a single nucleotide thereof. ASOs can be synthesized and / or modified by methods known in the art. Modifications at one nucleotide are independent of modifications at another nucleotide.

[0093] A "subunit" of an oligonucleotide refers to a nucleotide (or nucleotide analog) unit that contains a purine or pyrimidine base-pairing moiety. The term can refer to a nucleotide unit with or without an attached internucleotide bond, however, when referring to a "charged subunit", the charge is typically within the internucleotide bond (e.g., a phosphodiester or phosphorothioate bond or a cationic bond).

[0094] The purine or pyrimidine base pairing moiety (also referred to herein simply as "nucleobase", "base", or "base") can be adenine, cytosine, guanine, uracil, thymine, or inosine. Also included are bases such as pyridin-4-one, pyridin-2-one, phenyl, pseudouracil, 2,4,6-trimethoxybenzene, 3-methyluracil, dihydrouridine, naphthyl, aminophenyl, 5-alkylcytidine (e.g., 5-methylcytidine), 5-alkyluridine (e.g., ribothymidine), 5-halouridine (e.g., 5-bromouridine), or 6-aza pyrimidines or 6-alkyl pyrimidines (e.g., 6-methyluridine), propyne, queosine, 2-thiouridine, 4-thiouridine, wybutosine, wybutoxosine, 4-acetylcytidine, 5-(carboxyhydroxymethyl)uridine, 5'-carboxymethylaminomethyl-2-thiouridine, 5-carboxymethylaminomethyluridine, β-D-galactosylqueosine, 1-methyladenosine, 1-methylinosine, 2,2-dimethylguanosine, 3-methylcytidine, 2-methyladenosine, 2-methylguanosine, N6-methyladenosine, 7-methylguanosine, 5-methoxyaminomethyl-2-thiouridine, 5-methylaminomethyluridine, 5-methylcarbonylmethyluridine, 5-methoxyuridine, 5-methyl-2-thiouridine, 2-methylthio-N6-isopentenyladenosine, β-D-mannosylqueosine, uridine-5-hydroxyacetate, 2-thiocytidine, threonine derivatives, and others (Burgin et al., 1996, Biochemistry, 35:14090; Uhlman and Peyman, supra). In this regard, a "modified base" refers to a nucleobase other than adenine (A), guanine (G), cytosine (C), thymine (T), and uracil (U), as illustrated above; such bases can be used at any position in the antisense molecule. Those skilled in the art will understand that Ts and Us are interchangeable depending on the use of the oligomer. For example, for other antisense chemicals, such as more RNA-like 2'-O-methyl antisense oligonucleotides, the T base can be shown as U.

[0095] The term "targeting sequence" is a sequence in an oligomer or oligomer analogue that is complementary (and additionally meant to be substantially complementary) to a "target sequence" in an RNA genome. The entire sequence or only a portion of an antisense oligomer can be complementary to the target sequence. For example, in an oligomer having 20 - 30 bases, about 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, or 29 bases can be the targeting sequence that is complementary to the target region. Typically, the targeting sequence is formed by contiguous bases in the oligomer, but can alternatively be formed by non - contiguous sequences that, when placed together (e.g., from opposite ends of the oligomer), constitute a sequence spanning the target sequence.

[0096] A "targeting sequence" can have "near" or "substantial" complementarity to the target sequence and still function for the purposes of this disclosure, that is, still be "complementary". Preferably, the oligomer analogue compounds employed in this disclosure have at most one mismatch in 10 nucleotides with the target sequence and preferably at most one mismatch in 20 nucleotides. Alternatively, the antisense oligomers used have at least 90% sequence identity and preferably at least 95% sequence identity with the exemplary targeting sequences as specified herein.

[0097] An "amino acid subunit" or "amino acid residue" can refer to an α - amino acid residue (CO CHR - NH -) or a β - or other amino acid residue (e.g., -CO-(CH2)nCHR - NH-), where R is a side chain (which can include hydrogen), and n is from 1 to 7, preferably 1 to 4.

[0098] The term "naturally occurring amino acid" refers to the amino acids present in proteins found in nature, such as the 20 (L) - amino acids used during protein biosynthesis and other amino acids such as 4 - hydroxyproline, hydroxylysine, desmosine, isodesmosine, homocysteine, citrulline, and ornithine. The term "non - natural amino acid" refers to those amino acids that are not present in proteins found in nature. Examples include β - alanine (β - Ala), 6 - aminohexanoic acid (Ahx), and 6 - aminopentanoic acid. Additional examples of "non - natural amino acids" include, but are not limited to, (D) - amino acids, norleucine, norvaline, p - fluorophenylalanine, ethionine, etc., known to those of skill in the art.

[0099] The term "target sequence" refers to a portion of the target RNA to which an oligonucleotide or antisense agent is directed; that is, the sequence to which the oligonucleotide will hybridize via Watson-Crick base pairing of complementary sequences. In embodiments, the target sequence can be a contiguous region of a pre-mRNA that includes both intronic target sequences and exonic target sequences. In embodiments, the target sequence will consist of only intronic sequences or exonic sequences.

[0100] When hybridization occurs in an antiparallel configuration, the target and the targeting sequence are described as "complementary" to each other. The targeting sequence can have "near" or "substantial" complementarity to the target sequence and still function for the purposes of the present disclosure; that is, it can still be functionally "complementary". In certain embodiments, the oligonucleotide can have at most one mismatch in 10 nucleotides and preferably at most one mismatch in 20 nucleotides with the target sequence. Alternatively, the oligonucleotide can have at least 90% sequence identity and preferably at least 95% sequence identity with the exemplary antisense targeting sequences described herein.

[0101] An oligonucleotide "specifically hybridizes" to a target polynucleotide if it hybridizes to the target under physiological conditions where the Tm is substantially greater than 45 °C, preferably at least 50 °C and typically 60 °C - 80 °C or higher. Such hybridization preferably corresponds to stringent hybridization conditions. At a given ionic strength and pH, the Tm is the temperature at which 50% of the target sequence hybridizes to the complementary polynucleotide. Again, such hybridization can occur where the antisense oligomer has "near" or "substantial" complementarity as well as exact complementarity to the target sequence.

[0102] An "endonuclease-resistant" oligomeric molecule (oligomer) is a molecule whose backbone is substantially resistant to endonuclease cleavage in non-hybridized or hybridized form; by extracellular and intracellular endonucleases common in vivo; that is, under normal nuclease conditions, the oligomer shows little or no endonuclease cleavage in the body to which it is exposed.

[0103] An "effective amount" or "therapeutically effective amount" refers to the amount of a therapeutic agent (such as an UNC13A cryptic splice variant inhibitor) administered to a mammalian subject in a single dose or as part of a series of doses that is effective to produce a desired therapeutic effect. For an antisense oligonucleotide, such an effect is typically achieved by inhibiting the translation or natural splicing processing of a selected target sequence. An "effective amount" for an UNC13A cryptic exon splice variant mRNA also refers to an amount that is effective to modulate the expression of the UNC13A cryptic exon splice variant protein.

[0104] The terms "inhibit" or "inhibitor" refer to a direct or indirect alteration, interference, reduction, downregulation, blockage, inhibition, elimination, or degradation of the expression, amount, or activity of a target gene, target protein, or signaling pathway, relative to (1) a control, endogenous, or reference target or pathway or (2) the absence of the target or pathway, where the alteration, interference, reduction, downregulation, blockage, inhibition, elimination, or degradation is statistically, biologically, or clinically significant. The terms "inhibit" or "inhibitor" include gene "knockout" and gene "knockdown" methods, such as by chromosomal editing.

[0105] "Treatment" of an individual or cell is any type of intervention provided as a means of altering the natural course of a disease or pathology in the individual or cell. Treatment includes, but is not limited to, administration of, for example, a pharmaceutical composition, and can be prophylactic or after the onset of a pathological event or after contact with a pathogen. Treatment includes any desired effect on the symptoms or pathology of a disease or condition associated with inflammation, along with other effects described herein.

[0106] Also included is "prophylactic" treatment, which can be aimed at reducing the rate of progression of the disease or condition being treated, delaying the onset of the disease or condition, or reducing the severity of its onset. "Treatment" or "prevention" does not necessarily indicate complete eradication, cure, or prevention of the disease or condition or its associated symptoms.

[0107] I. UNC13A cryptic exon splicing variants

[0108] On the one hand, the present disclosure provides novel UNC13A-targeting oligonucleotides that reduce cryptic splicing variants including cryptic exons between exons 20 and 21. These cryptic exons are not present in wild-type UNC13A from neuronal cell nuclei and are not present in any known UNC13A isoforms. The cryptic exons are derived from intron 20-21 of the UNC13A gene (SEQ ID NO:4). Depletion of TDP-43 introduced two alternative 3' splice acceptors in intron 20-21, one introduced at chr19:17642591 (ΔΨ = 0.05184) and the other at chr19:17642541 (ΔΨ = 0.48865). An alternative 5' splice donor was also introduced at chr19:17642414 (ΔΨ = 0.772). The chr19:17642541 3' splice acceptor (which is used more frequently than the chr19:17642591 3' splice acceptor) and the alternative 5' splice donor gave rise to a 128bp cryptic exon (“cryptic exon #1”) having the nucleotide sequence shown in SEQ ID NO:5. The UNC13A cryptic exon #1 variant contains the nucleotide sequence shown in SEQ ID NO:7, which encodes a protein comprising the amino acid sequence shown in SEQ ID NO:8. The chr19:17642591 3' splice acceptor and the alternative 5' splice donor gave rise to a 179bp cryptic exon (“cryptic exon #2”) having the nucleotide sequence shown in SEQ ID NO:6. The UNC13A cryptic exon #2 variant contains the nucleotide sequence shown in SEQ ID NO:9, which encodes a protein comprising the amino acid sequence shown in SEQ ID NO:10.

[0109] Compared to normal controls, the expression level of the UNC13A cryptic exon #1 splicing variant was significantly increased in the frontal cortex of patients with frontotemporal lobar degeneration (FTLD-TDP) with TDP-43 inclusions. The UNC13A cryptic exon #1 splicing variant was also detected in disease-related tissues of ALS patients. In an embodiment, the expression of UNC13A cryptic splicing variant #1 or UNC13A cryptic splicing variant #2 can be used as a biomarker for identifying a subject suffering from a TDP-43 proteinopathy (e.g., FTLD or ALS).

[0110] Once TDP-43 is depleted from the nucleus and accumulates in the cytoplasm, it becomes phosphorylated. Hyperphosphorylated TDP-43 (pTDP-43) is a key feature of the pathology of TDP-43 proteinopathies. The UNC13A cryptic exon #1 splicing variant is strongly correlated with the level of phosphorylated TDP-43 in patients with FTD / ALS. In an embodiment, the expression of UNC13A cryptic splicing variant #1 or UNC13A cryptic splicing variant #2 can be used as a biomarker for the level of phosphorylated TDP-43 in a subject.

[0111] A number of genetic mutations in intron 20-21 of UNC13A have been identified to promote the inclusion of the UNC13A cryptic exon upon TDP-43 depletion. Examples of such genetic mutations include rs12608932 (hg38 chr19:17,641,880A→C), rs12973192 (hg38 chr19:17,642,430C→G), rs56041637 (hg38chr19:17,642,033-17,642,056CATC 0-2 repeats→3-5CATC repeats), and rs62121687 (hg38chr19:17,642,351C→A). In addition, UNC13A genetic mutations that increase cryptic exon inclusion are associated with reduced survival in FTD-ALS patients. In an embodiment, the identification of a genetic mutation in intron 20-21 of UNC13A in a subject can be used as a biomarker for the inclusion of the UNC13A cryptic exon. In an embodiment, the identification of a genetic mutation in intron 20-21 of UNC13A in a subject with a TDP-43 proteinopathy (e.g., FTD, ALS) can be used as a biomarker for reduced survival.

[0112] Table 1: UNC13A and TDP-43 sequences

[0113]

[0114]

[0115]

[0116]

[0117]

[0118]

[0119]

[0120]

[0121]

[0122]

[0123]

[0124]

[0125]

[0126]

[0127] II. UNC13A Splice-Blocking Antisense Oligonucleotides

[0128] In some embodiments, provided herein are antisense oligonucleotides that target the cryptic exon of UNC13A. In an embodiment, the cryptic exon is obtained from intron 20-21 of the UNC13A gene. In an embodiment, the cryptic exon comprises SEQ ID NO:5 or SEQ ID NO:6. In an embodiment, the UNC13 cryptic exon splicing variant comprises the polynucleotide sequence of SEQ ID NO:7 or SEQ ID NO:9. In an embodiment, the UNC13A cryptic exon splicing variant comprises the amino acid sequence of SEQ ID NO:8 or SEQ ID NO:10. In some embodiments, the ASO reduces the level of the UNC13A cryptic exon splicing variant.

[0129] In an embodiment, the antisense oligonucleotide comprises an inhibitory nucleic acid. The inhibitory nucleic acid can be an antisense oligonucleotide, siRNA, shRNA, miRNA, double-stranded RNA (dsRNA), or esiRNA. In an embodiment, the inhibitory nucleic acid comprises an antisense oligonucleotide complementary to: the exon 20 splice donor site region in the pre-processed mRNA encoding UNC13A; the cryptic exon splice acceptor site region in the pre-processed mRNA encoding UNC13A; the cryptic exon splice donor site region in the pre-processed mRNA encoding UNC13A; or the exon 21 splice acceptor site region in the pre-processed mRNA encoding UNC13A. In an embodiment, the exon 20 splice donor site region comprises SEQ ID NO:800 or consists thereof. In an embodiment, the cryptic exon splice acceptor site region comprises SEQ ID NO:801 or consists thereof. In an embodiment, the cryptic exon splice donor site region comprises SEQ ID NO:802 or consists thereof. In an embodiment, the exon 21 splice acceptor site comprises SEQ ID NO:803 or consists thereof.

[0130] In an embodiment, the antisense oligonucleotide selectively inhibits the expression or activity of the UNC13A cryptic exon splice variant on full-length UNC13A (wild type) or other variants thereof (i.e., variants that do not contain the cryptic exon from introns 20-21, such as SEQ ID NO:5 or SEQ ID NO:6).

[0131] Table 2A: Antisense oligonucleotide sequences

[0132]

[0133]

[0134]

[0135]

[0136]

[0137]

[0138]

[0139]

[0140]

[0141]

[0142]

[0143]

[0144]

[0145]

[0146]

[0147]

[0148]

[0149]

[0150] Table 2B: Additional antisense oligonucleotide sequences

[0151]

[0152]

[0153]

[0154]

[0155]

[0156]

[0157]

[0158]

[0159]

[0160]

[0161]

[0162]

[0163]

[0164]

[0165] The antisense oligonucleotides (ASOs) provided herein may alternatively be designated by the sequence ID numbers and ASO numbers provided in Tables 2A and 2B. The ASO numbers MTx_ASO_(number) and only said number may be used interchangeably. For example, MTx_ASO_280 may alternatively be referred to as ASO 280. In some cases, the ASO number refers to the region targeted by the corresponding ASO. For example, MTx_ASO_Region_4_Child_1 (SEQ ID NO:961) targets Region 4 (SEQ ID NO:703) of the cryptic exon as described herein.

[0166] In some embodiments, the antisense oligonucleotide is complementary to the cryptic exon of UNC13 mRNA. In some embodiments, the antisense oligonucleotide comprises or consists of the sequences shown in SEQ ID NOs: 20-40. In some embodiments, the antisense oligonucleotide has a length of about 15-50 bases, preferably 16 bases, 17 bases, 18 bases, 19 bases, 20 bases, 21 bases, 22 bases, 23 bases, 24 bases or 25 bases. In some embodiments, the antisense oligonucleotide comprises a sequence that is at least 80%, 85%, 90%, 95%, 97% or 100% identical to any one of SEQ ID NOs: 20-40. In some embodiments, the antisense oligonucleotide is a modified antisense oligonucleotide. In some embodiments, the antisense oligonucleotide comprises a 2′OMe antisense oligonucleotide, a 2′-O-methoxyethyl antisense oligonucleotide, a phosphorothioate antisense oligonucleotide or an LNA antisense oligonucleotide. In some embodiments, the antisense oligonucleotide blocks the inclusion of the cryptic exon by steric hindrance of the cryptic 5′ donor splice site or the 3′ cryptic acceptor site. The antisense oligonucleotide can block the inclusion of the cryptic exon by modulating the rate of RNA polymerase II. The antisense oligonucleotide can block the inclusion of the cryptic exon by modifying the structure of the mRNA. The antisense oligonucleotide can block the inclusion of the cryptic exon by other known or unknown mechanisms capable of blocking the inclusion of the cryptic exon. In some embodiments, the antisense oligonucleotide increases UNC13A protein expression. In some embodiments, the antisense oligonucleotide reduces the inclusion of the UNC13A cryptic exon.

[0167] In some embodiments, the antisense oligonucleotide is complementary to the cryptic exon of UNC13 mRNA. In some embodiments, the antisense oligonucleotide comprises or consists of the sequences shown in SEQ ID NOs: 20 - 355 or SEQ ID NOs: 810 - 957. In some embodiments, the antisense oligonucleotide has a length of about 15 - 50 bases, preferably 16 bases, 17 bases, 18 bases, 19 bases, 20 bases, 21 bases, 22 bases, 23 bases, 24 bases, or 25 bases. In some embodiments, the antisense oligonucleotide comprises a sequence that is at least 80%, 85%, 90%, 95%, 97%, or 100% identical to any one of SEQ ID NOs: 20 - 355 or SEQ ID NOs: 810 - 975. In some embodiments, the antisense oligonucleotide is a modified antisense oligonucleotide. In some embodiments, the antisense oligonucleotide comprises a 2'OMe antisense oligonucleotide, a 2'O-methoxyethyl antisense oligonucleotide, a phosphorothioate antisense oligonucleotide, or an LNA antisense oligonucleotide. In some embodiments, the antisense oligonucleotide blocks the inclusion of the cryptic exon by steric hindrance of the cryptic 5' donor splice site or the 3' cryptic acceptor site. The antisense oligonucleotide can block the inclusion of the cryptic exon by modulating the rate of RNA polymerase II. The antisense oligonucleotide can block the inclusion of the cryptic exon by modifying the structure of the mRNA. The antisense oligonucleotide can block the inclusion of the cryptic exon by other known or unknown mechanisms capable of blocking the inclusion of the cryptic exon. In some embodiments, the antisense oligonucleotide increases UNC13A protein expression. In some embodiments, the antisense oligonucleotide reduces the inclusion of the UNC13A cryptic exon.

[0168] In some embodiments, the antisense oligonucleotide is complementary to the cryptic exon of UNC13 mRNA. In some embodiments, the antisense oligonucleotide comprises or consists of the sequences shown in SEQ ID NO: 20-355 or SEQ ID NO: 400-638 or SEQ ID NO: 810-957. In some embodiments, the antisense oligonucleotide has a length of about 15-50 bases, preferably 16 bases, 17 bases, 18 bases, 19 bases, 20 bases, 21 bases, 22 bases, 23 bases, 24 bases or 25 bases. In some embodiments, the antisense oligonucleotide comprises a sequence that is at least 80%, 85%, 90%, 95%, 97% or 100% identical to any one of SEQ ID NO: 20-355 or SEQ ID NO: 400-638 or SEQ ID NO: 810-957. In some embodiments, the antisense oligonucleotide is a modified antisense oligonucleotide. In some embodiments, the antisense oligonucleotide comprises a 2′OMe antisense oligonucleotide, a 2′-O-methoxyethyl antisense oligonucleotide, a phosphorothioate antisense oligonucleotide or a LNA antisense oligonucleotide. In some embodiments, the antisense oligonucleotide blocks the inclusion of the cryptic exon by steric hindrance of the cryptic 5′ donor splice site or the 3′ cryptic acceptor site. The antisense oligonucleotide can block the inclusion of the cryptic exon by modulating the rate of RNA polymerase II. The antisense oligonucleotide can block the inclusion of the cryptic exon by modifying the structure of the mRNA. The antisense oligonucleotide can block the inclusion of the cryptic exon by other known or unknown mechanisms capable of blocking the inclusion of the cryptic exon. In some embodiments, the antisense oligonucleotide increases UNC13A protein expression. In some embodiments, the antisense oligonucleotide reduces the inclusion of the UNC13A cryptic exon.

[0169] In some embodiments, the antisense oligonucleotide is complementary to the cryptic exon of UNC13 mRNA. In some embodiments, the antisense oligonucleotide comprises or consists of the sequences shown in SEQ ID NO: 20 - 355, 400 - 638, 810 - 959, or 961 - 1332. In some embodiments, the antisense oligonucleotide has a length of about 13 - 50 bases, such as 13 bases, 14 bases, 15 bases, 16 bases, 17 bases, 18 bases, 19 bases, 20 bases, 21 bases, 22 bases, 23 bases, 24 bases, or 25 bases. In some embodiments, the antisense oligonucleotide comprises a sequence that is at least 80%, 85%, 90%, 95%, 97%, or 100% identical to any one of SEQ ID NO: 20 - 355, 400 - 638, 810 - 959, or 961 - 1332. In some embodiments, the antisense oligonucleotide is a modified antisense oligonucleotide. In some embodiments, the antisense oligonucleotide comprises a 2'OMe antisense oligonucleotide, a 2'O-methoxyethyl antisense oligonucleotide, a phosphorothioate antisense oligonucleotide, or an LNA antisense oligonucleotide. In some embodiments, the antisense oligonucleotide blocks the inclusion of the cryptic exon by steric hindrance of the cryptic 5'splice donor site or the 3'cryptic acceptor site. The antisense oligonucleotide can block the inclusion of the cryptic exon by modulating the rate of RNA polymerase II. The antisense oligonucleotide can block the inclusion of the cryptic exon by modifying the structure of the mRNA. The antisense oligonucleotide can block the inclusion of the cryptic exon by other known or unknown mechanisms capable of blocking the inclusion of the cryptic exon. In some embodiments, in some embodiments, the antisense oligonucleotide increases UNC13A protein expression. In some embodiments, the antisense oligonucleotide reduces the inclusion of the UNC13A cryptic exon.

[0170] In some embodiments, the antisense oligonucleotide is complementary to the 5' end of the cryptic exon of UNC13 mRNA. In some embodiments, the antisense oligonucleotide targets region 1 of UNC13A mRNA. In some embodiments, the antisense oligonucleotide is complementary to SEQ ID NO:700. In some embodiments, the antisense oligonucleotide comprises or consists of the sequence shown in SEQ ID NO:20 or SEQ ID NO:38. In some embodiments, the antisense oligonucleotide has a length of about 15-50 bases, preferably 16 bases, 17 bases, 18 bases, 19 bases, 20 bases, 21 bases, 22 bases, 23 bases, 24 bases or 25 bases. In some embodiments, the antisense oligonucleotide comprises a sequence that is at least 80%, 85%, 90%, 95%, 97% or 100% identical to either SEQ ID NO:20 or SEQ ID NO:38. In some embodiments, the antisense oligonucleotide is a modified antisense oligonucleotide. In some embodiments, the antisense oligonucleotide comprises a 2'-OMe antisense oligonucleotide, a 2'-O-methoxyethyl antisense oligonucleotide, a phosphorothioate antisense oligonucleotide or an LNA antisense oligonucleotide. In some embodiments, the antisense oligonucleotide blocks the inclusion of the cryptic exon by steric hindrance of cryptic 5' donor site splicing. The antisense oligonucleotide can block the inclusion of the cryptic exon by modulating the rate of RNA polymerase II. The antisense oligonucleotide can block the inclusion of the cryptic exon by modifying the structure of the mRNA. The antisense oligonucleotide can block the inclusion of the cryptic exon by other known or unknown mechanisms capable of blocking the inclusion of the cryptic exon. In some embodiments, the antisense oligonucleotide increases UNC13A protein expression. In some embodiments, the antisense oligonucleotide reduces the inclusion of the UNC13A cryptic exon.

[0171] In some embodiments, the antisense oligonucleotide is complementary to the 5' end of the cryptic exon of UNC13 mRNA. In some embodiments, the antisense oligonucleotide targets region 1 of UNC13A mRNA. In some embodiments, the antisense oligonucleotide is complementary to SEQ ID NO:700. In some embodiments, the antisense oligonucleotide comprises or consists of the sequences shown in SEQ ID NOs: 41-55 or SEQ ID NOs: 810-825. In some embodiments, the antisense oligonucleotide is about 15-50 bases in length, preferably 16 bases, 17 bases, 18 bases, 19 bases, 20 bases, 21 bases, 22 bases, 23 bases, 24 bases or 25 bases. In some embodiments, the antisense oligonucleotide comprises or consists of a sequence that is at least 80%, 85%, 90%, 95%, 97% or 100% identical to any one of SEQ ID NOs: 41-SEQ ID NO: 55 or SEQ ID NOs: 810-825. In some embodiments, the antisense oligonucleotide is a modified antisense oligonucleotide. In some embodiments, the antisense oligonucleotide comprises a 2'-OMe antisense oligonucleotide, a 2'-O-methoxyethyl antisense oligonucleotide, a phosphorothioate antisense oligonucleotide or a LNA antisense oligonucleotide. In some embodiments, the antisense oligonucleotide blocks the inclusion of the cryptic exon by steric hindrance of splicing at the cryptic 5' donor site. The antisense oligonucleotide can block the inclusion of the cryptic exon by modulating the rate of RNA polymerase II. The antisense oligonucleotide can block the inclusion of the cryptic exon by modifying the structure of the mRNA. The antisense oligonucleotide can block the inclusion of the cryptic exon by other known or unknown mechanisms capable of blocking the inclusion of the cryptic exon. In some embodiments, the antisense oligonucleotide increases UNC13A protein expression. In some embodiments, the antisense oligonucleotide reduces the inclusion of the UNC13A cryptic exon.

[0172] In some embodiments, the antisense oligonucleotide is complementary to the 5' end of a cryptic exon of UNC13 mRNA. In some embodiments, the antisense oligonucleotide targets Region 1 of UNC13A mRNA. In some embodiments, the antisense oligonucleotide is complementary to SEQ ID NO:700. In some embodiments, the antisense oligonucleotide comprises or consists of the sequences shown in SEQ ID NOs: 311-325 or SEQ ID NOs: 910-929. In some embodiments, the antisense oligonucleotide has a length of about 15-50 bases, preferably 16 bases, 17 bases, 18 bases, 19 bases, 20 bases, 21 bases, 22 bases, 23 bases, 24 bases, or 25 bases. In some embodiments, the antisense oligonucleotide comprises or consists of a sequence that is at least 80%, 85%, 90%, 95%, 97%, or 100% identical to any one of SEQ ID NOs: 311-325 or SEQ ID NOs: 910-929. In some embodiments, the antisense oligonucleotide is a modified antisense oligonucleotide. In some embodiments, the antisense oligonucleotide comprises a 2'-OMe antisense oligonucleotide, a 2'-O-methoxyethyl antisense oligonucleotide, a phosphorothioate antisense oligonucleotide, or a LNA antisense oligonucleotide. In some embodiments, the antisense oligonucleotide blocks the inclusion of the cryptic exon by steric hindrance of cryptic 5' donor site splicing. The antisense oligonucleotide can block the inclusion of the cryptic exon by modulating the rate of RNA polymerase II. The antisense oligonucleotide can block the inclusion of the cryptic exon by modifying the structure of the mRNA. The antisense oligonucleotide can block the inclusion of the cryptic exon by other known or unknown mechanisms capable of blocking the inclusion of the cryptic exon. In some embodiments, the antisense oligonucleotide increases UNC13A protein expression. In some embodiments, the antisense oligonucleotide reduces the inclusion of the UNC13A cryptic exon.

[0173] In some embodiments, the antisense oligonucleotide is complementary to an intervening region of a cryptic exon of UNC13 mRNA located between the 5' splice site and the 3' splice site. In some embodiments, the antisense oligonucleotide targets region 2 of UNC13A mRNA. In some embodiments, the antisense oligonucleotide is complementary to SEQ ID NO:701. In some embodiments, the antisense oligonucleotide comprises or consists of the sequences shown in SEQ ID NOs: 34-37. In some embodiments, the antisense oligonucleotide is about 15-50 bases in length, preferably 16 bases, 17 bases, 18 bases, 19 bases, 20 bases, 21 bases, 22 bases, 23 bases, 24 bases, or 25 bases. In some embodiments, the antisense oligonucleotide comprises or consists of a sequence that is at least 80%, 85%, 90%, 95%, 97%, or 100% identical to any one of SEQ ID NOs: 34-37. In some embodiments, the antisense oligonucleotide is a modified antisense oligonucleotide. In some embodiments, the antisense oligonucleotide comprises a 2'OMe antisense oligonucleotide, a 2'-O-methoxyethyl antisense oligonucleotide, a phosphorothioate antisense oligonucleotide, or a LNA antisense oligonucleotide. In some embodiments, the antisense oligonucleotide blocks the inclusion of the cryptic exon by steric hindrance of the cryptic 5' donor site splicing or the 3' cryptic acceptor site. The antisense oligonucleotide can block the inclusion of the cryptic exon by modulating the rate of RNA polymerase II. The antisense oligonucleotide can block the inclusion of the cryptic exon by modifying the structure of the mRNA. The antisense oligonucleotide can block the inclusion of the cryptic exon by other known or unknown mechanisms capable of blocking the inclusion of the cryptic exon. In some embodiments, the antisense oligonucleotide increases UNC13A protein expression. In some embodiments, the antisense oligonucleotide reduces the inclusion of the UNC13A cryptic exon.

[0174] In some embodiments, the antisense oligonucleotide is complementary to the intervening region of the cryptic exon of UNC13 mRNA located between the 5' splice site and the 3' splice site. In some embodiments, the antisense oligonucleotide targets region 2 of UNC13A mRNA. In some embodiments, the antisense oligonucleotide is complementary to SEQ ID NO:701. In some embodiments, the antisense oligonucleotide comprises or consists of the sequences shown in SEQ ID NO:251-265 or SEQ ID NO:877-890. In some embodiments, the antisense oligonucleotide is about 15-50 bases in length, preferably 16 bases, 17 bases, 18 bases, 19 bases, 20 bases, 21 bases, 22 bases, 23 bases, 24 bases or 25 bases. In some embodiments, the antisense oligonucleotide comprises a sequence that is at least 80%, 85%, 90%, 95%, 97% or 100% identical to any one of SEQ ID NO:251-265 or SEQ ID NO:877-890. In some embodiments, the antisense oligonucleotide is a modified antisense oligonucleotide. In some embodiments, the antisense oligonucleotide comprises a 2' OMe antisense oligonucleotide, a 2'-O-methoxyethyl antisense oligonucleotide, a phosphorothioate antisense oligonucleotide or a LNA antisense oligonucleotide. In some embodiments, the antisense oligonucleotide blocks the inclusion of the cryptic exon by steric hindrance of the cryptic 5' donor site splicing or the 3' cryptic acceptor site. The antisense oligonucleotide can block the inclusion of the cryptic exon by modulating the rate of RNA polymerase II. The antisense oligonucleotide can block the inclusion of the cryptic exon by modifying the structure of the mRNA. The antisense oligonucleotide can block the inclusion of the cryptic exon by other known or unknown mechanisms capable of blocking the inclusion of the cryptic exon. In some embodiments, the antisense oligonucleotide increases UNC13A protein expression. In some embodiments, the antisense oligonucleotide reduces the inclusion of the UNC13A cryptic exon.

[0175] In some embodiments, the antisense oligonucleotide is complementary to an intermediate region of a cryptic exon of UNC13 mRNA located between the 5' splice site and the 3' splice site. In some embodiments, the antisense oligonucleotide targets region 2 of UNC13A mRNA. In some embodiments, the antisense oligonucleotide is complementary to SEQ ID NO:701. In some embodiments, the antisense oligonucleotide comprises or consists of the sequences shown in SEQ ID NO:266-280 or SEQ ID NO:891-894. In some embodiments, the antisense oligonucleotide is about 15-50 bases in length, preferably 16 bases, 17 bases, 18 bases, 19 bases, 20 bases, 21 bases, 22 bases, 23 bases, 24 bases, or 25 bases. In some embodiments, the antisense oligonucleotide comprises a sequence that is at least 80%, 85%, 90%, 95%, 97%, or 100% identical to any one of SEQ ID NO:266-280 or SEQ ID NO:891-894. In some embodiments, the antisense oligonucleotide is a modified antisense oligonucleotide. In some embodiments, the antisense oligonucleotide comprises a 2'OMe antisense oligonucleotide, a 2'-O-methoxyethyl antisense oligonucleotide, a phosphorothioate antisense oligonucleotide, or a LNA antisense oligonucleotide. In some embodiments, the antisense oligonucleotide blocks the inclusion of the cryptic exon by steric hindrance of the cryptic 5' donor splice site or the 3' cryptic acceptor site. The antisense oligonucleotide can block the inclusion of the cryptic exon by modulating the rate of RNA polymerase II. The antisense oligonucleotide can block the inclusion of the cryptic exon by modifying the structure of the mRNA. The antisense oligonucleotide can block the inclusion of the cryptic exon by other known or unknown mechanisms capable of blocking the inclusion of the cryptic exon. In some embodiments, the antisense oligonucleotide increases UNC13A protein expression. In some embodiments, the antisense oligonucleotide reduces the inclusion of the UNC13A cryptic exon.

[0176] In some embodiments, the antisense oligonucleotide is complementary to an intervening region of a cryptic exon of UNC13 mRNA located between the 5' splice site and the 3' splice site. In some embodiments, the antisense oligonucleotide targets region 2 of UNC13A mRNA. In some embodiments, the antisense oligonucleotide is complementary to SEQ ID NO:701. In some embodiments, the antisense oligonucleotide comprises or consists of the sequences shown in SEQ ID NOs: 281-295 or SEQ ID NOs: 895-899. In some embodiments, the antisense oligonucleotide is about 15-50 bases in length, preferably 16 bases, 17 bases, 18 bases, 19 bases, 20 bases, 21 bases, 22 bases, 23 bases, 24 bases, or 25 bases. In some embodiments, the antisense oligonucleotide comprises a sequence that is at least 80%, 85%, 90%, 95%, 97%, or 100% identical to any one of SEQ ID NOs: 281-295 or SEQ ID NOs: 895-899. In some embodiments, the antisense oligonucleotide is a modified antisense oligonucleotide. In some embodiments, the antisense oligonucleotide comprises a 2' OMe antisense oligonucleotide, a 2'-O-methoxyethyl antisense oligonucleotide, a phosphorothioate antisense oligonucleotide, or a LNA antisense oligonucleotide. In some embodiments, the antisense oligonucleotide blocks the inclusion of the cryptic exon by steric hindrance of the cryptic 5' donor site splicing or the 3' cryptic acceptor site. The antisense oligonucleotide can block the inclusion of the cryptic exon by modulating the rate of RNA polymerase II. The antisense oligonucleotide can block the inclusion of the cryptic exon by modifying the structure of the mRNA. The antisense oligonucleotide can block the inclusion of the cryptic exon by other known or unknown mechanisms capable of blocking the inclusion of the cryptic exon. In some embodiments, the antisense oligonucleotide increases UNC13A protein expression. In some embodiments, the antisense oligonucleotide reduces the inclusion of the UNC13A cryptic exon.

[0177] In some embodiments, the antisense oligonucleotide is complementary to an intervening region of a cryptic exon of UNC13 mRNA located between the 5' splice site and the 3' splice site. In some embodiments, the antisense oligonucleotide targets region 2 of UNC13A mRNA. In some embodiments, the antisense oligonucleotide is complementary to SEQ ID NO:701. In some embodiments, the antisense oligonucleotide comprises or consists of the sequences shown in SEQ ID NO:296 - 310 or SEQ ID NO:900 - 909. In some embodiments, the antisense oligonucleotide is about 15 - 50 bases in length, preferably 16 bases, 17 bases, 18 bases, 19 bases, 20 bases, 21 bases, 22 bases, 23 bases, 24 bases, or 25 bases. In some embodiments, the antisense oligonucleotide comprises a sequence that is at least 80%, 85%, 90%, 95%, 97%, or 100% identical to any one of SEQ ID NO:296 - 310 or SEQ ID NO:900 - 909. In some embodiments, the antisense oligonucleotide is a modified antisense oligonucleotide. In some embodiments, the antisense oligonucleotide comprises a 2'OMe antisense oligonucleotide, a 2'O-methoxyethyl antisense oligonucleotide, a phosphorothioate antisense oligonucleotide, or an LNA antisense oligonucleotide. In some embodiments, the antisense oligonucleotide blocks the inclusion of the cryptic exon by steric hindrance of the cryptic 5' donor site splicing or the 3' cryptic acceptor site. The antisense oligonucleotide can block the inclusion of the cryptic exon by modulating the rate of RNA polymerase II. The antisense oligonucleotide can block the inclusion of the cryptic exon by modifying the structure of the mRNA. The antisense oligonucleotide can block the inclusion of the cryptic exon by other known or unknown mechanisms capable of blocking the inclusion of the cryptic exon. In some embodiments, the antisense oligonucleotide increases UNC13A protein expression. In some embodiments, the antisense oligonucleotide reduces the inclusion of the UNC13A cryptic exon.

[0178] In some embodiments, the antisense oligonucleotide is complementary to the middle region of the cryptic exon of UNC13 mRNA located between the 5' splice site and the 3' splice site. In some embodiments, the antisense oligonucleotide targets region 2 of UNC13A mRNA or region 3 of UNC13A mRNA. In some embodiments, the antisense oligonucleotide is complementary to SEQ ID NO:701 or 702. In some embodiments, the antisense oligonucleotide comprises or consists of the sequence shown in SEQ ID NO:40. In some embodiments, the antisense oligonucleotide is about 15-50 bases in length, preferably 16 bases, 17 bases, 18 bases, 19 bases, 20 bases, 21 bases, 22 bases, 23 bases, 24 bases or 25 bases. In some embodiments, the antisense oligonucleotide comprises a sequence that is at least 80%, 85%, 90%, 95%, 97% or 100% identical to any one of those in SEQ ID NO:40. In some embodiments, the antisense oligonucleotide is a modified antisense oligonucleotide. In some embodiments, the antisense oligonucleotide comprises a 2' OMe antisense oligonucleotide, a 2'-O-methoxyethyl antisense oligonucleotide, a phosphorothioate antisense oligonucleotide or a LNA antisense oligonucleotide. In some embodiments, the antisense oligonucleotide blocks the inclusion of the cryptic exon by steric hindrance of the cryptic 5' donor splice site or the 3' cryptic acceptor site. The antisense oligonucleotide can block the inclusion of the cryptic exon by modulating the rate of RNA polymerase II. The antisense oligonucleotide can block the inclusion of the cryptic exon by modifying the structure of the mRNA. The antisense oligonucleotide can block the inclusion of the cryptic exon by other known or unknown mechanisms capable of blocking the inclusion of the cryptic exon. In some embodiments, the antisense oligonucleotide increases UNC13A protein expression. In some embodiments, the antisense oligonucleotide reduces the inclusion of the UNC13A cryptic exon.

[0179] In some embodiments, the antisense oligonucleotide is complementary to an intermediate region of a cryptic exon of UNC13 mRNA located between the 5' splice site and the 3' splice site. In some embodiments, the antisense oligonucleotide targets region 2 of UNC13A mRNA or region 3 of UNC13A mRNA. In some embodiments, the antisense oligonucleotide is complementary to SEQ ID NO:701 or 702. In some embodiments, the antisense oligonucleotide comprises or consists of the sequences shown in SEQ ID NO:341-355 or SEQ ID NO:934-957. In some embodiments, the antisense oligonucleotide has a length of about 15-50 bases, preferably 16 bases, 17 bases, 18 bases, 19 bases, 20 bases, 21 bases, 22 bases, 23 bases, 24 bases or 25 bases. In some embodiments, the antisense oligonucleotide comprises a sequence that is at least 80%, 85%, 90%, 95%, 97% or 100% identical to any one of SEQ ID NO:341-355 or SEQ ID NO:934-957. In some embodiments, the antisense oligonucleotide is a modified antisense oligonucleotide. In some embodiments, the antisense oligonucleotide comprises a 2'OMe antisense oligonucleotide, a 2'-O-methoxyethyl antisense oligonucleotide, a phosphorothioate antisense oligonucleotide or an LNA antisense oligonucleotide. In some embodiments, the antisense oligonucleotide blocks the inclusion of the cryptic exon by steric hindrance of the cryptic 5' donor site splicing or the 3' cryptic acceptor site. The antisense oligonucleotide can block the inclusion of the cryptic exon by modulating the rate of RNA polymerase II. The antisense oligonucleotide can block the inclusion of the cryptic exon by modifying the structure of the mRNA. The antisense oligonucleotide can block the inclusion of the cryptic exon by other known or unknown mechanisms capable of blocking the inclusion of the cryptic exon. In some embodiments, the antisense oligonucleotide increases UNC13A protein expression. In some embodiments, the antisense oligonucleotide reduces the inclusion of the UNC13A cryptic exon.

[0180] In some embodiments, the antisense oligonucleotide is complementary to the middle region of a cryptic exon of UNC13 mRNA located between the 5' splice site and the 3' splice site. In some embodiments, the antisense oligonucleotide targets region 2 of UNC13A mRNA. In some embodiments, the antisense oligonucleotide is complementary to SEQ ID NO:701. In some embodiments, the antisense oligonucleotide comprises or consists of the sequences shown in SEQ ID NOs:1116-1332. In some embodiments, the antisense oligonucleotide has a length of about 13-50 bases, such as 13 bases, 14 bases, 15 bases, 16 bases, 17 bases, 18 bases, 19 bases, 20 bases, 21 bases, 22 bases, 23 bases, 24 bases or 25 bases. In some embodiments, the antisense oligonucleotide comprises a sequence that is at least 80%, 85%, 90%, 95%, 97% or 100% identical to any one of SEQ ID NOs:1116-1332. In some embodiments, the antisense oligonucleotide is a modified antisense oligonucleotide. In some embodiments, the antisense oligonucleotide comprises a 2'OMe antisense oligonucleotide, a 2'-O-methoxyethyl antisense oligonucleotide, a phosphorothioate antisense oligonucleotide or a LNA antisense oligonucleotide. In some embodiments, the antisense oligonucleotide blocks the inclusion of the cryptic exon by steric hindrance of the cryptic 5' donor site splicing or the 3' cryptic acceptor site. The antisense oligonucleotide can block the inclusion of the cryptic exon by modulating the rate of RNA polymerase II. The antisense oligonucleotide can block the inclusion of the cryptic exon by modifying the structure of the mRNA. The antisense oligonucleotide can block the inclusion of the cryptic exon by other known or unknown mechanisms capable of blocking the inclusion of the cryptic exon. In some embodiments, the antisense oligonucleotide increases UNC13A mRNA expression. In some embodiments, the antisense oligonucleotide increases UNC13A protein expression. In some embodiments, the antisense oligonucleotide reduces the inclusion of the UNC13A cryptic exon.

[0181] In some embodiments, the antisense oligonucleotide is complementary to the 3'-end of the cryptic exon of UNC13 mRNA. In some embodiments, the antisense oligonucleotide targets region 3 of UNC13A mRNA. In some embodiments, the antisense oligonucleotide is complementary to SEQ ID NO:702. In some embodiments, the antisense oligonucleotide comprises or consists of the sequences shown in SEQ ID NOs: 30 - 33. In some embodiments, the antisense oligonucleotide has a length of about 15 - 50 bases, preferably 16 bases, 17 bases, 18 bases, 19 bases, 20 bases, 21 bases, 22 bases, 23 bases, 24 bases, or 25 bases. In some embodiments, the antisense oligonucleotide comprises a sequence that is at least 80%, 85%, 90%, 95%, 97%, or 100% identical to any one of SEQ ID NOs: 21 - 31, SEQ ID NO: 33, or SEQ ID NO: 39. In some embodiments, the antisense oligonucleotide is a modified antisense oligonucleotide. In some embodiments, the antisense oligonucleotide comprises a 2'-OMe antisense oligonucleotide, a 2'-O-methoxyethyl antisense oligonucleotide, a phosphorothioate antisense oligonucleotide, or an LNA antisense oligonucleotide. In some embodiments, the antisense oligonucleotide blocks the inclusion of the cryptic exon by steric hindrance at the 3'-cryptic acceptor site. The antisense oligonucleotide can block the inclusion of the cryptic exon by modulating the rate of RNA polymerase II. The antisense oligonucleotide can block the inclusion of the cryptic exon by modifying the structure of the mRNA. The antisense oligonucleotide can block the inclusion of the cryptic exon by other known or unknown mechanisms capable of blocking the inclusion of the cryptic exon. In some embodiments, the antisense oligonucleotide increases UNC13A protein expression. In some embodiments, the antisense oligonucleotide reduces the inclusion of the UNC13A cryptic exon.

[0182] In some embodiments, the antisense oligonucleotide is complementary to the 3' end of the cryptic exon of UNC13 mRNA. In some embodiments, the antisense oligonucleotide targets region 3 of UNC13A mRNA. In some embodiments, the antisense oligonucleotide is complementary to SEQ ID NO:702. In some embodiments, the antisense oligonucleotide comprises or consists of the sequences shown in SEQ ID NOs: 191-205 or SEQ ID NOs: 851-860. In some embodiments, the antisense oligonucleotide has a length of about 15-50 bases, preferably 16 bases, 17 bases, 18 bases, 19 bases, 20 bases, 21 bases, 22 bases, 23 bases, 24 bases or 25 bases. In some embodiments, the antisense oligonucleotide comprises a sequence that is at least 80%, 85%, 90%, 95%, 97% or 100% identical to any one of SEQ ID NOs: 191-205 or SEQ ID NOs: 851-860. In some embodiments, the antisense oligonucleotide is a modified antisense oligonucleotide. In some embodiments, the antisense oligonucleotide comprises a 2'-OMe antisense oligonucleotide, a 2'-O-methoxyethyl antisense oligonucleotide, a phosphorothioate antisense oligonucleotide or an LNA antisense oligonucleotide. In some embodiments, the antisense oligonucleotide blocks the inclusion of the cryptic exon by steric hindrance at the 3' cryptic acceptor site. The antisense oligonucleotide can block the inclusion of the cryptic exon by modulating the rate of RNA polymerase II. The antisense oligonucleotide can block the inclusion of the cryptic exon by modifying the structure 3 of the mRNA. The antisense oligonucleotide can block the inclusion of the cryptic exon by other known or unknown mechanisms capable of blocking the inclusion of the cryptic exon. In some embodiments, the antisense oligonucleotide increases UNC13A protein expression. In some embodiments, the antisense oligonucleotide reduces the inclusion of the UNC13A cryptic exon.

[0183] In some embodiments, the antisense oligonucleotide is complementary to the 3'-end of the cryptic exon of UNC13 mRNA. In some embodiments, the antisense oligonucleotide targets region 3 of UNC13A mRNA. In some embodiments, the antisense oligonucleotide is complementary to SEQ ID NO:702. In some embodiments, the antisense oligonucleotide comprises or consists of the sequences shown in SEQ ID NOs: 206-220 or SEQ ID NOs: 861-863. In some embodiments, the antisense oligonucleotide is about 15-50 bases in length, preferably 16 bases, 17 bases, 18 bases, 19 bases, 20 bases, 21 bases, 22 bases, 23 bases, 24 bases or 25 bases. In some embodiments, the antisense oligonucleotide comprises a sequence that is at least 80%, 85%, 90%, 95%, 97% or 100% identical to any one of SEQ ID NOs: 206-220 or SEQ ID NOs: 861-863. In some embodiments, the antisense oligonucleotide is a modified antisense oligonucleotide. In some embodiments, the antisense oligonucleotide comprises a 2'-OMe antisense oligonucleotide, a 2'-O-methoxyethyl antisense oligonucleotide, a phosphorothioate antisense oligonucleotide or a LNA antisense oligonucleotide. In some embodiments, the antisense oligonucleotide blocks the inclusion of the cryptic exon by steric hindrance at the 3'-cryptic acceptor site. The antisense oligonucleotide can block the inclusion of the cryptic exon by modulating the rate of RNA polymerase II. The antisense oligonucleotide can block the inclusion of the cryptic exon by modifying the structure of the mRNA. The antisense oligonucleotide can block the inclusion of the cryptic exon by other known or unknown mechanisms capable of blocking the inclusion of the cryptic exon. In some embodiments, the antisense oligonucleotide increases UNC13A protein expression. In some embodiments, the antisense oligonucleotide reduces the inclusion of the UNC13A cryptic exon.

[0184] In some embodiments, the antisense oligonucleotide is complementary to the 3'-end of the cryptic exon of UNC13 mRNA. In some embodiments, the antisense oligonucleotide targets region 3 of UNC13A mRNA. In some embodiments, the antisense oligonucleotide is complementary to SEQ ID NO:702. In some embodiments, the antisense oligonucleotide comprises or consists of the sequences shown in SEQ ID NOs: 221-234 or SEQ ID NOs: 864-866. In some embodiments, the antisense oligonucleotide is about 15-50 bases in length, preferably 16 bases, 17 bases, 18 bases, 19 bases, 20 bases, 21 bases, 22 bases, 23 bases, 24 bases or 25 bases. In some embodiments, the antisense oligonucleotide comprises a sequence that is at least 80%, 85%, 90%, 95%, 97% or 100% identical to any one of SEQ ID NOs: 221-225 or SEQ ID NOs: 864-866. In some embodiments, the antisense oligonucleotide is a modified antisense oligonucleotide. In some embodiments, the antisense oligonucleotide comprises a 2'-OMe antisense oligonucleotide, a 2'-O-methoxyethyl antisense oligonucleotide, a phosphorothioate antisense oligonucleotide or an LNA antisense oligonucleotide. In some embodiments, the antisense oligonucleotide blocks the inclusion of the cryptic exon by steric hindrance at the 3'-cryptic acceptor site. The antisense oligonucleotide may block the inclusion of the cryptic exon by modulating the rate of RNA polymerase II. The antisense oligonucleotide may block the inclusion of the cryptic exon by modifying the structure of the mRNA. The antisense oligonucleotide may block the inclusion of the cryptic exon by other known or unknown mechanisms capable of blocking the inclusion of the cryptic exon. In some embodiments, the antisense oligonucleotide increases UNC13A protein expression. In some embodiments, the antisense oligonucleotide reduces the inclusion of the UNC13A cryptic exon.

[0185] In some embodiments, the antisense oligonucleotide is complementary to the 3' end of the cryptic exon of UNC13 mRNA. In some embodiments, the antisense oligonucleotide targets region 3 of UNC13A mRNA. In some embodiments, the antisense oligonucleotide is complementary to SEQ ID NO:702. In some embodiments, the antisense oligonucleotide comprises or consists of the sequences shown in SEQ ID NOs: 236-250 or SEQ ID NOs: 867-876. In some embodiments, the antisense oligonucleotide is about 15-50 bases in length, preferably 16 bases, 17 bases, 18 bases, 19 bases, 20 bases, 21 bases, 22 bases, 23 bases, 24 bases, or 25 bases. In some embodiments, the antisense oligonucleotide comprises a sequence that is at least 80%, 85%, 90%, 95%, 97%, or 100% identical to any one of SEQ ID NOs: 236-250 or SEQ ID NOs: 867-876. In some embodiments, the antisense oligonucleotide is a modified antisense oligonucleotide. In some embodiments, the antisense oligonucleotide comprises a 2' OMe antisense oligonucleotide, a 2'-O-methoxyethyl antisense oligonucleotide, a phosphorothioate antisense oligonucleotide, or a LNA antisense oligonucleotide. In some embodiments, the antisense oligonucleotide blocks the inclusion of the cryptic exon by steric hindrance at the 3' cryptic acceptor site. The antisense oligonucleotide can block the inclusion of the cryptic exon by modulating the rate of RNA polymerase II. The antisense oligonucleotide can block the inclusion of the cryptic exon by modifying the structure of the mRNA. The antisense oligonucleotide can block the inclusion of the cryptic exon by other known or unknown mechanisms capable of blocking the inclusion of the cryptic exon. In some embodiments, the antisense oligonucleotide increases UNC13A protein expression. In some embodiments, the antisense oligonucleotide reduces the inclusion of the UNC13A cryptic exon.

[0186] In some embodiments, the antisense oligonucleotide is complementary to the 3' end of the cryptic exon of UNC13 mRNA. In some embodiments, the antisense oligonucleotide targets region 4 of UNC13A mRNA. In some embodiments, the antisense oligonucleotide is complementary to SEQ ID NO:703. In some embodiments, the antisense oligonucleotide comprises or consists of the sequences shown in SEQ ID NO:21-29 or SEQ ID NO:39. In some embodiments, the antisense oligonucleotide has a length of about 15-50 bases, preferably 16 bases, 17 bases, 18 bases, 19 bases, 20 bases, 21 bases, 22 bases, 23 bases, 24 bases or 25 bases. In some embodiments, the antisense oligonucleotide comprises a sequence that is at least 80%, 85%, 90%, 95%, 97% or 100% identical to any one of SEQ ID NO:21-29 or SEQ ID NO:39. In some embodiments, the antisense oligonucleotide is a modified antisense oligonucleotide. In some embodiments, the antisense oligonucleotide comprises a 2'OMe antisense oligonucleotide, a 2'-O-methoxyethyl antisense oligonucleotide, a phosphorothioate antisense oligonucleotide or an LNA antisense oligonucleotide. In some embodiments, the antisense oligonucleotide blocks the inclusion of the cryptic exon by steric hindrance at the 3' cryptic acceptor site. The antisense oligonucleotide can block the inclusion of the cryptic exon by modulating the rate of RNA polymerase II. The antisense oligonucleotide can block the inclusion of the cryptic exon by modifying the structure of the mRNA. The antisense oligonucleotide can block the inclusion of the cryptic exon by other known or unknown mechanisms capable of blocking the inclusion of the cryptic exon. In some embodiments, the antisense oligonucleotide increases UNC13A protein expression. In some embodiments, the antisense oligonucleotide reduces the inclusion of the UNC13A cryptic exon.

[0187] In some embodiments, the antisense oligonucleotide is complementary to the 3' end of the cryptic exon of UNC13 mRNA. In some embodiments, the antisense oligonucleotide targets region 4 of UNC13A mRNA. In some embodiments, the antisense oligonucleotide is complementary to SEQ ID NO: 703. In some embodiments, the antisense oligonucleotide comprises or consists of the sequences shown in SEQ ID NOs: 56 - 70 or SEQ ID NOs: 826 - 834. In some embodiments, the antisense oligonucleotide has a length of about 15 - 50 bases, preferably 16 bases, 17 bases, 18 bases, 19 bases, 20 bases, 21 bases, 22 bases, 23 bases, 24 bases, or 25 bases. In some embodiments, the antisense oligonucleotide comprises a sequence that is at least 80%, 85%, 90%, 95%, 97%, or 100% identical to any one of SEQ ID NOs: 56 - 70 or SEQ ID NOs: 826 - 834. In some embodiments, the antisense oligonucleotide is a modified antisense oligonucleotide. In some embodiments, the antisense oligonucleotide comprises a 2'-OMe antisense oligonucleotide, a 2'-O-methoxyethyl antisense oligonucleotide, a phosphorothioate antisense oligonucleotide, or a LNA antisense oligonucleotide. In some embodiments, the antisense oligonucleotide blocks the inclusion of the cryptic exon by steric hindrance at the 3' cryptic acceptor site. The antisense oligonucleotide can block the inclusion of the cryptic exon by modulating the rate of RNA polymerase II. The antisense oligonucleotide can block the inclusion of the cryptic exon by modifying the structure of the mRNA. The antisense oligonucleotide can block the inclusion of the cryptic exon by other known or unknown mechanisms capable of blocking the inclusion of the cryptic exon. In some embodiments, the antisense oligonucleotide increases UNC13A protein expression. In some embodiments, the antisense oligonucleotide reduces the inclusion of the UNC13A cryptic exon.

[0188] In some embodiments, the antisense oligonucleotide is complementary to the 3' end of the cryptic exon of UNC13 mRNA. In some embodiments, the antisense oligonucleotide targets region 4 of UNC13A mRNA. In some embodiments, the antisense oligonucleotide is complementary to SEQ ID NO:703. In some embodiments, the antisense oligonucleotide comprises or consists of the sequences shown in SEQ ID NOs:71-85 or SEQ ID NOs:835-837. In some embodiments, the antisense oligonucleotide has a length of about 15-50 bases, preferably 16 bases, 17 bases, 18 bases, 19 bases, 20 bases, 21 bases, 22 bases, 23 bases, 24 bases or 25 bases. In some embodiments, the antisense oligonucleotide comprises a sequence that is at least 80%, 85%, 90%, 95%, 97% or 100% identical to any one of SEQ ID NOs:71-85 or SEQ ID NOs:835-837. In some embodiments, the antisense oligonucleotide is a modified antisense oligonucleotide. In some embodiments, the antisense oligonucleotide comprises a 2'-OMe antisense oligonucleotide, a 2'-O-methoxyethyl antisense oligonucleotide, a phosphorothioate antisense oligonucleotide or a LNA antisense oligonucleotide. In some embodiments, the antisense oligonucleotide blocks the inclusion of the cryptic exon by steric hindrance at the 3' cryptic acceptor site. The antisense oligonucleotide can block the inclusion of the cryptic exon by modulating the rate of RNA polymerase II. The antisense oligonucleotide can block the inclusion of the cryptic exon by modifying the structure of the mRNA. The antisense oligonucleotide can block the inclusion of the cryptic exon by other known or unknown mechanisms capable of blocking the inclusion of the cryptic exon. In some embodiments, the antisense oligonucleotide increases UNC13A protein expression. In some embodiments, the antisense oligonucleotide reduces the inclusion of the UNC13A cryptic exon.

[0189] In some embodiments, the antisense oligonucleotide is complementary to the 3'-end of the cryptic exon of UNC13 mRNA. In some embodiments, the antisense oligonucleotide targets region 4 of UNC13A mRNA. In some embodiments, the antisense oligonucleotide is complementary to SEQ ID NO:703. In some embodiments, the antisense oligonucleotide comprises or consists of the sequences shown in SEQ ID NOs: 86-100. In some embodiments, the antisense oligonucleotide has a length of about 15-50 bases, preferably 16 bases, 17 bases, 18 bases, 19 bases, 20 bases, 21 bases, 22 bases, 23 bases, 24 bases, or 25 bases. In some embodiments, the antisense oligonucleotide comprises a sequence that is at least 80%, 85%, 90%, 95%, 97%, or 100% identical to any one of SEQ ID NOs: 86-100. In some embodiments, the antisense oligonucleotide is a modified antisense oligonucleotide. In some embodiments, the antisense oligonucleotide comprises a 2'-OMe antisense oligonucleotide, a 2'-O-methoxyethyl antisense oligonucleotide, a phosphorothioate antisense oligonucleotide, or a LNA antisense oligonucleotide. In some embodiments, the antisense oligonucleotide blocks the inclusion of the cryptic exon by steric hindrance at the 3'-cryptic acceptor site. The antisense oligonucleotide can block the inclusion of the cryptic exon by modulating the rate of RNA polymerase II. The antisense oligonucleotide can block the inclusion of the cryptic exon by modifying the structure of the mRNA. The antisense oligonucleotide can block the inclusion of the cryptic exon by other known or unknown mechanisms capable of blocking the inclusion of the cryptic exon. In some embodiments, the antisense oligonucleotide increases UNC13A protein expression. In some embodiments, the antisense oligonucleotide reduces the inclusion of the UNC13A cryptic exon.

[0190] In some embodiments, the antisense oligonucleotide is complementary to the 3' end of the cryptic exon of UNC13 mRNA. In some embodiments, the antisense oligonucleotide targets region 4 of UNC13A mRNA. In some embodiments, the antisense oligonucleotide is complementary to SEQ ID NO: 703. In some embodiments, the antisense oligonucleotide comprises or consists of the sequences shown in SEQ ID NOs: 101 - 115. In some embodiments, the antisense oligonucleotide is about 15 - 50 bases in length, preferably 16 bases, 17 bases, 18 bases, 19 bases, 20 bases, 21 bases, 22 bases, 23 bases, 24 bases or 25 bases. In some embodiments, the antisense oligonucleotide comprises a sequence that is at least 80%, 85%, 90%, 95%, 97% or 100% identical to any one of SEQ ID NOs: 101 - 115. In some embodiments, the antisense oligonucleotide is a modified antisense oligonucleotide. In some embodiments, the antisense oligonucleotide comprises a 2'-OMe antisense oligonucleotide, a 2'-O-methoxyethyl antisense oligonucleotide, a phosphorothioate antisense oligonucleotide or a LNA antisense oligonucleotide. In some embodiments, the antisense oligonucleotide blocks the inclusion of the cryptic exon by steric hindrance at the 3' cryptic acceptor site. The antisense oligonucleotide can block the inclusion of the cryptic exon by modulating the rate of RNA polymerase II. The antisense oligonucleotide can block the inclusion of the cryptic exon by modifying the structure of the mRNA. The antisense oligonucleotide can block the inclusion of the cryptic exon by other known or unknown mechanisms capable of blocking the inclusion of the cryptic exon. In some embodiments, the antisense oligonucleotide increases UNC13A protein expression. In some embodiments, the antisense oligonucleotide reduces the inclusion of the UNC13A cryptic exon.

[0191] In some embodiments, the antisense oligonucleotide is complementary to the 3' end of the cryptic exon of UNC13 mRNA. In some embodiments, the antisense oligonucleotide targets region 4 of UNC13A mRNA. In some embodiments, the antisense oligonucleotide is complementary to SEQ ID NO:703. In some embodiments, the antisense oligonucleotide comprises or consists of the sequences shown in SEQ ID NOs:116-130. In some embodiments, the antisense oligonucleotide has a length of about 15-50 bases, preferably 16 bases, 17 bases, 18 bases, 19 bases, 20 bases, 21 bases, 22 bases, 23 bases, 24 bases or 25 bases. In some embodiments, the antisense oligonucleotide comprises a sequence that is at least 80%, 85%, 90%, 95%, 97% or 100% identical to any one of SEQ ID NOs:116-130. In some embodiments, the antisense oligonucleotide is a modified antisense oligonucleotide. In some embodiments, the antisense oligonucleotide comprises a 2'OMe antisense oligonucleotide, a 2'-O-methoxyethyl antisense oligonucleotide, a phosphorothioate antisense oligonucleotide or an LNA antisense oligonucleotide. In some embodiments, the antisense oligonucleotide blocks the inclusion of the cryptic exon by steric hindrance at the 3' cryptic acceptor site. The antisense oligonucleotide can block the inclusion of the cryptic exon by modulating the rate of RNA polymerase II. The antisense oligonucleotide can block the inclusion of the cryptic exon by modifying the structure of the mRNA. The antisense oligonucleotide can block the inclusion of the cryptic exon by other known or unknown mechanisms capable of blocking the inclusion of the cryptic exon. In some embodiments, the antisense oligonucleotide increases UNC13A protein expression. In some embodiments, the antisense oligonucleotide reduces the inclusion of the UNC13A cryptic exon.

[0192] In some embodiments, the antisense oligonucleotide is complementary to the 3' end of the cryptic exon of UNC13 mRNA. In some embodiments, the antisense oligonucleotide targets region 4 of UNC13A mRNA. In some embodiments, the antisense oligonucleotide is complementary to SEQ ID NO:703. In some embodiments, the antisense oligonucleotide comprises or consists of the sequences shown in SEQ ID NOs:131 - 145. In some embodiments, the antisense oligonucleotide has a length of about 15 - 50 bases, preferably 16 bases, 17 bases, 18 bases, 19 bases, 20 bases, 21 bases, 22 bases, 23 bases, 24 bases or 25 bases. In some embodiments, the antisense oligonucleotide comprises a sequence that is at least 80%, 85%, 90%, 95%, 97% or 100% identical to any one of SEQ ID NOs:131 - 145. In some embodiments, the antisense oligonucleotide is a modified antisense oligonucleotide. In some embodiments, the antisense oligonucleotide comprises a 2' - OMe antisense oligonucleotide, a 2'-O-methoxyethyl antisense oligonucleotide, a phosphorothioate antisense oligonucleotide or a LNA antisense oligonucleotide. In some embodiments, the antisense oligonucleotide blocks the inclusion of the cryptic exon by steric hindrance at the 3' cryptic acceptor site. The antisense oligonucleotide can block the inclusion of the cryptic exon by modulating the rate of RNA polymerase II. The antisense oligonucleotide can block the inclusion of the cryptic exon by modifying the structure of the mRNA. The antisense oligonucleotide can block the inclusion of the cryptic exon by other known or unknown mechanisms capable of blocking the inclusion of the cryptic exon. In some embodiments, the antisense oligonucleotide increases UNC13A protein expression. In some embodiments, the antisense oligonucleotide reduces the inclusion of the UNC13A cryptic exon.

[0193] In some embodiments, the antisense oligonucleotide is complementary to the 3' end of the cryptic exon of UNC13 mRNA. In some embodiments, the antisense oligonucleotide targets region 4 of UNC13A mRNA. In some embodiments, the antisense oligonucleotide is complementary to SEQ ID NO: 703. In some embodiments, the antisense oligonucleotide comprises or consists of the sequence shown in SEQ ID NOs: 56 - 70. In some embodiments, the antisense oligonucleotide has a length of about 146 - 160 bases, preferably 16 bases, 17 bases, 18 bases, 19 bases, 20 bases, 21 bases, 22 bases, 23 bases, 24 bases, or 25 bases. In some embodiments, the antisense oligonucleotide comprises a sequence that is at least 80%, 85%, 90%, 95%, 97%, or 100% identical to any one of SEQ ID NOs: 146 - 160. In some embodiments, the antisense oligonucleotide is a modified antisense oligonucleotide. In some embodiments, the antisense oligonucleotide comprises a 2'-OMe antisense oligonucleotide, a 2'-O-methoxyethyl antisense oligonucleotide, a phosphorothioate antisense oligonucleotide, or an LNA antisense oligonucleotide. In some embodiments, the antisense oligonucleotide blocks the inclusion of the cryptic exon by steric hindrance at the 3' cryptic acceptor site. The antisense oligonucleotide can block the inclusion of the cryptic exon by modulating the rate of RNA polymerase II. The antisense oligonucleotide can block the inclusion of the cryptic exon by modifying the structure of the mRNA. The antisense oligonucleotide can block the inclusion of the cryptic exon by other known or unknown mechanisms capable of blocking the inclusion of the cryptic exon. In some embodiments, the antisense oligonucleotide increases UNC13A protein expression. In some embodiments, the antisense oligonucleotide reduces the inclusion of the UNC13A cryptic exon.

[0194] In some embodiments, the antisense oligonucleotide is complementary to the 3' end of the cryptic exon of UNC13 mRNA. In some embodiments, the antisense oligonucleotide targets region 4 of UNC13A mRNA. In some embodiments, the antisense oligonucleotide is complementary to SEQ ID NO:703. In some embodiments, the antisense oligonucleotide comprises or consists of the sequences shown in SEQ ID NOs: 161-175 or SEQ ID NOs: 838-840. In some embodiments, the antisense oligonucleotide is about 15-50 bases in length, preferably 16 bases, 17 bases, 18 bases, 19 bases, 20 bases, 21 bases, 22 bases, 23 bases, 24 bases or 25 bases. In some embodiments, the antisense oligonucleotide comprises a sequence that is at least 80%, 85%, 90%, 95%, 97% or 100% identical to any one of SEQ ID NOs: 161-175 or SEQ ID NOs: 838-840. In some embodiments, the antisense oligonucleotide is a modified antisense oligonucleotide. In some embodiments, the antisense oligonucleotide comprises a 2'-OMe antisense oligonucleotide, a 2'-O-methoxyethyl antisense oligonucleotide, a phosphorothioate antisense oligonucleotide or a LNA antisense oligonucleotide. In some embodiments, the antisense oligonucleotide blocks the inclusion of the cryptic exon by steric hindrance at the 3' cryptic acceptor site. The antisense oligonucleotide can block the inclusion of the cryptic exon by modulating the rate of RNA polymerase II. The antisense oligonucleotide can block the inclusion of the cryptic exon by modifying the structure of the mRNA. The antisense oligonucleotide can block the inclusion of the cryptic exon by other known or unknown mechanisms capable of blocking the inclusion of the cryptic exon. In some embodiments, the antisense oligonucleotide increases UNC13A protein expression. In some embodiments, the antisense oligonucleotide reduces the inclusion of the UNC13A cryptic exon.

[0195] In some embodiments, the antisense oligonucleotide is complementary to the 3' end of the cryptic exon of UNC13 mRNA. In some embodiments, the antisense oligonucleotide targets region 4 of UNC13A mRNA. In some embodiments, the antisense oligonucleotide is complementary to SEQ ID NO:703. In some embodiments, the antisense oligonucleotide comprises or consists of the sequences shown in SEQ ID NOs: 176-190 or SEQ ID NOs: 841-850. In some embodiments, the antisense oligonucleotide has a length of about 15-50 bases, preferably 16 bases, 17 bases, 18 bases, 19 bases, 20 bases, 21 bases, 22 bases, 23 bases, 24 bases, or 25 bases. In some embodiments, the antisense oligonucleotide comprises a sequence that is at least 80%, 85%, 90%, 95%, 97%, or 100% identical to any one of SEQ ID NOs: 176-190 or SEQ ID NOs: 841-850. In some embodiments, the antisense oligonucleotide is a modified antisense oligonucleotide. In some embodiments, the antisense oligonucleotide comprises a 2'-OMe antisense oligonucleotide, a 2'-O-methoxyethyl antisense oligonucleotide, a phosphorothioate antisense oligonucleotide, or a LNA antisense oligonucleotide. In some embodiments, the antisense oligonucleotide blocks the inclusion of the cryptic exon by steric hindrance at the 3' cryptic acceptor site. The antisense oligonucleotide can block the inclusion of the cryptic exon by modulating the rate of RNA polymerase II. The antisense oligonucleotide can block the inclusion of the cryptic exon by modifying the structure of the mRNA. The antisense oligonucleotide can block the inclusion of the cryptic exon by other known or unknown mechanisms capable of blocking the inclusion of the cryptic exon. In some embodiments, the antisense oligonucleotide increases UNC13A protein expression. In some embodiments, the antisense oligonucleotide reduces the inclusion of the UNC13A cryptic exon.

[0196] In some embodiments, the antisense oligonucleotide is complementary to the 3' end of the cryptic exon of UNC13 mRNA. In some embodiments, the antisense oligonucleotide targets region 4 of UNC13A mRNA. In some embodiments, the antisense oligonucleotide is complementary to SEQ ID NO:703. In some embodiments, the antisense oligonucleotide comprises or consists of the sequences shown in SEQ ID NOs: 326-340 or SEQ ID NOs: 930-933. In some embodiments, the antisense oligonucleotide is about 15-50 bases in length, preferably 16 bases, 17 bases, 18 bases, 19 bases, 20 bases, 21 bases, 22 bases, 23 bases, 24 bases or 25 bases. In some embodiments, the antisense oligonucleotide comprises a sequence that is at least 80%, 85%, 90%, 95%, 97% or 100% identical to any one of SEQ ID NOs: 326-340 or SEQ ID NOs: 930-933. In some embodiments, the antisense oligonucleotide is a modified antisense oligonucleotide. In some embodiments, the antisense oligonucleotide comprises a 2' OMe antisense oligonucleotide, a 2'-O-methoxyethyl antisense oligonucleotide, a phosphorothioate antisense oligonucleotide or a LNA antisense oligonucleotide. In some embodiments, the antisense oligonucleotide blocks the inclusion of the cryptic exon by steric hindrance at the 3' cryptic acceptor site. The antisense oligonucleotide can block the inclusion of the cryptic exon by modulating the rate of RNA polymerase II. The antisense oligonucleotide can block the inclusion of the cryptic exon by modifying the structure of the mRNA. The antisense oligonucleotide can block the inclusion of the cryptic exon by other known or unknown mechanisms capable of blocking the inclusion of the cryptic exon. In some embodiments, the antisense oligonucleotide increases UNC13A protein expression. In some embodiments, the antisense oligonucleotide reduces the inclusion of the UNC13A cryptic exon.

[0197] In some embodiments, the antisense oligonucleotide is complementary to the 3' end of the cryptic exon of UNC13 mRNA. In some embodiments, the antisense oligonucleotide targets region 4 of UNC13A mRNA. In some embodiments, the antisense oligonucleotide is complementary to SEQ ID NO:703. In some embodiments, the antisense oligonucleotide comprises or consists of the sequences shown in SEQ ID NOs: 961-1115. In some embodiments, the antisense oligonucleotide has a length of about 13-50 bases, such as 13 bases, 14 bases, 15 bases, 16 bases, 17 bases, 18 bases, 19 bases, 20 bases, 21 bases, 22 bases, 23 bases, 24 bases, or 25 bases. In some embodiments, the antisense oligonucleotide comprises a sequence that is at least 80%, at least 85%, at least 90%, at least 95%, at least 97%, or 100% identical to any one of SEQ ID NOs: 961-1115. In some embodiments, the antisense oligonucleotide is a modified antisense oligonucleotide. In some embodiments, the antisense oligonucleotide comprises a 2'-OMe antisense oligonucleotide, a 2'-O-methoxyethyl antisense oligonucleotide, a phosphorothioate antisense oligonucleotide, or an LNA antisense oligonucleotide. In some embodiments, the antisense oligonucleotide blocks the inclusion of the cryptic exon by steric hindrance at the 3' cryptic acceptor site. The antisense oligonucleotide can block the inclusion of the cryptic exon by modulating the rate of RNA polymerase II. The antisense oligonucleotide can block the inclusion of the cryptic exon by modifying the structure of the mRNA. The antisense oligonucleotide can block the inclusion of the cryptic exon by other known or unknown mechanisms capable of blocking the inclusion of the cryptic exon. In some embodiments, the antisense oligonucleotide increases UNC13A mRNA expression. In some embodiments, the antisense oligonucleotide increases UNC13A protein expression. In some embodiments, the antisense oligonucleotide reduces the inclusion of the UNC13A cryptic exon.

[0198] In some embodiments, the antisense oligonucleotide is complementary to the 3' end of the cryptic exon of UNC13 mRNA. In some embodiments, the antisense oligonucleotide targets region 1, 2, 3, or 4 of UNC13A mRNA. In some embodiments, the antisense oligonucleotide is complementary to any one of SEQ ID NOs: 700-703. In some embodiments, the antisense oligonucleotide is complementary to the cryptic exon of UNC13 mRNA. In some embodiments, the antisense oligonucleotide comprises or consists of the sequences shown in SEQ ID NOs: 400-638. In some embodiments, the antisense oligonucleotide has a length of about 15-50 bases, preferably 16 bases, 17 bases, 18 bases, 19 bases, 20 bases, 21 bases, 22 bases, 23 bases, 24 bases, or 25 bases. In some embodiments, the antisense oligonucleotide comprises a sequence that is at least 80%, 85%, 90%, 95%, 97%, or 100% identical to any one of SEQ ID NOs: 400-638. In some embodiments, the antisense oligonucleotide is a modified antisense oligonucleotide. In some embodiments, the antisense oligonucleotide comprises a 2'OMe antisense oligonucleotide, a 2'-O-methoxyethyl antisense oligonucleotide, a phosphorothioate antisense oligonucleotide, or an LNA antisense oligonucleotide. In some embodiments, the antisense oligonucleotide blocks the inclusion of the cryptic exon by steric hindrance of the cryptic 5' donor splice site or the 3' cryptic acceptor site. The antisense oligonucleotide can block the inclusion of the cryptic exon by modulating the rate of RNA polymerase II. The antisense oligonucleotide can block the inclusion of the cryptic exon by modifying the structure of the mRNA. The antisense oligonucleotide can block the inclusion of the cryptic exon by other known or unknown mechanisms capable of blocking the inclusion of the cryptic exon. In some embodiments, the antisense oligonucleotide increases UNC13A protein expression. In some embodiments, the antisense oligonucleotide reduces the inclusion of the UNC13A cryptic exon.

[0199] In some embodiments, the antisense oligonucleotide has at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 97% or 100% complementarity with any one of SEQ ID NO:5-10 or SEQ ID NO:700-703. In some embodiments, in some embodiments, the antisense oligonucleotide has a length of about 15-50 bases, preferably 16 bases, 17 bases, 18 bases, 19 bases, 20 bases, 21 bases, 22 bases, 23 bases, 24 bases or 25 bases. In some embodiments, the antisense oligonucleotide is a modified antisense oligonucleotide. In some embodiments, the antisense oligonucleotide comprises a 2'OMe antisense oligonucleotide, a 2'-O-methoxyethyl antisense oligonucleotide, a phosphorothioate antisense oligonucleotide or a LNA antisense oligonucleotide. In some embodiments, the antisense oligonucleotide blocks the inclusion of a cryptic exon by steric hindrance of the 5' donor splice site or the 3' cryptic acceptor site. The antisense oligonucleotide can block the inclusion of a cryptic exon by modulating the rate of RNA polymerase II. The antisense oligonucleotide can block the inclusion of a cryptic exon by modifying the structure of the mRNA. The antisense oligonucleotide can block the inclusion of a cryptic exon by other known or unknown mechanisms capable of blocking the inclusion of a cryptic exon. In some embodiments, the antisense oligonucleotide increases UNC13A protein expression. In some embodiments, the antisense oligonucleotide reduces the inclusion of the UNC13A cryptic exon.

[0200] In some aspects, provided herein is an antisense oligonucleotide that modulates UNC13A splicing, wherein the antisense oligonucleotide has a sequence complementary to the sequence of UNC13A mRNA, the sequence of the UNC13A mRNA being between the 5' splice donor site and the 3' splice acceptor site of a cryptic exon, the cryptic exon being between exons 20 and 21 of the UNC13A gene.

[0201] In some embodiments, the antisense oligonucleotide is complementary to the sequence shown in SEQ ID NO:1. In some embodiments, the antisense oligonucleotide has at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 95% or at least 98% complementarity with the sequence shown in SEQ ID NO:1.

[0202] In some embodiments, the antisense oligonucleotide is complementary to the sequence shown in any one of SEQ ID NOs: 700-703. In some embodiments, the antisense oligonucleotide is complementary to the sequence shown in SEQ ID NO: 701 or 703. In some embodiments, the antisense oligonucleotide has at least 50%, 60%, 70%, 80%, 90%, 95% or 98% complementarity with the sequence shown in SEQ ID NO: 701 or 703. In some embodiments, the antisense oligonucleotide is completely complementary to the sequence shown in SEQ ID NO: 701 or 703. In some embodiments, the antisense oligonucleotide is complementary to the sequence shown in any one of SEQ ID NOs: 700. In some embodiments, the antisense oligonucleotide has at least 50%, 60%, 70%, 80%, 90%, 95% or 98% complementarity with the sequence shown in SEQ ID NO: 700. In some embodiments, the antisense oligonucleotide is completely complementary to the sequence shown in SEQ ID NO: 700. In some embodiments, the antisense oligonucleotide is complementary to the sequence shown in any one of SEQ ID NOs: 701. In some embodiments, the antisense oligonucleotide has at least 50%, 60%, 70%, 80%, 90%, 95% or 98% complementarity with the sequence shown in SEQ ID NO: 701. In some embodiments, the antisense oligonucleotide is completely complementary to the sequence shown in SEQ ID NO: 701. In some embodiments, the antisense oligonucleotide is complementary to the sequence shown in any one of SEQ ID NOs: 702. In some embodiments, the antisense oligonucleotide has at least 50%, 60%, 70%, 80%, 90%, 95% or 98% complementarity with the sequence shown in SEQ ID NO: 702. In some embodiments, the antisense oligonucleotide is completely complementary to the sequence shown in SEQ ID NO: 702. In some embodiments, the antisense oligonucleotide is complementary to the sequence shown in any one of SEQ ID NOs: 703. In some embodiments, the antisense oligonucleotide has at least 50%, 60%, 70%, 80%, 90%, 95% or 98% complementarity with the sequence shown in SEQ ID NO: 703. In some embodiments, the antisense oligonucleotide is completely complementary to the sequence shown in SEQ ID NO: 703.

[0203] In some embodiments, the antisense oligonucleotide comprises at least 80%, at least 90%, at least 95%, at least 98% or 100% complementarity to the sequences shown in SEQ ID NOs: 700 - 703. In some embodiments, the antisense oligonucleotide comprises at least 80%, at least 90%, at least 95%, at least 98% or 100% complementarity to the sequence shown in SEQ ID NO: 701. In some embodiments, the antisense oligonucleotide comprises at least 80%, at least 90%, at least 95%, at least 98% or 100% complementarity to the sequence shown in SEQ ID NO: 703.

[0204] In some cases, the antisense oligonucleotide is complementary to the sequence of UNC13A mRNA, which sequence is located between the 5' splice donor site and the 3' splice acceptor site of a cryptic exon located between exons 20 and 21 of the UNC13A gene, wherein the sequence of UNC13A is uniquely sensitive to antisense oligonucleotide binding. In some cases, regions of UNC13A mRNA are identified as being more responsive than other regions. Here, it is determined that these regions are sensitive to a variety of factors, including but not limited to antisense oligonucleotide length, composition, and localization on UNC13A mRNA. The smallest regions of UNC13A mRNA are specifically targeted to maximize the activity of antisense oligonucleotides that regulate UNC13A mRNA splicing. In some embodiments, these highly responsive smallest regions of UNC13A mRNA are referred to as hotspots.

[0205] In some embodiments, the minimum region of UNC13A mRNA between the 5' splice donor site and the 3' splice acceptor site of a cryptic exon located between exons 20 and 21 of the UNC13A gene was identified as being highly responsive to binding with an antisense oligonucleotide or another similar nucleic acid probe. In some embodiments, an ASO that binds to such a minimum region is effective in preventing the inclusion of the UNC13A cryptic exon. In some embodiments, the minimum region is 3 to 30, 4 to 26, 5 to 22, 6 to 18, 7 to 14, or 8 to 10 nucleotides in length. In some embodiments, the minimum region is no more than 30, no more than 25, no more than 20, or no more than 15 nucleotides in length. In some embodiments, the minimum region is 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 nucleotides in length. In some embodiments, the minimum region is 18 nucleotides in length. In some embodiments, the minimum region is 29 nucleotides in length. In some embodiments, the minimum region comprises the sequence shown in any of SEQ ID NOs: 1339 - 1342 or a portion thereof. In some embodiments, the minimum region comprises the sequence shown in SEQ ID NO: 1339 or a portion thereof. In some embodiments, the minimum region comprises the sequence shown in SEQ ID NO: 1340 or a portion thereof. In some embodiments, the minimum region comprises the sequence shown in SEQ ID NO: 1341 or a portion thereof. In some embodiments, the minimum region comprises the sequence shown in SEQ ID NO: 1342 or a portion thereof. In some embodiments, binding of the region or a portion thereof to an antisense oligonucleotide enhances splicing regulatory activity. In some embodiments, binding of the minimum region or a portion thereof to an antisense oligonucleotide increases UNC13A mRNA expression. In some embodiments, binding of the minimum region or a portion thereof to an antisense oligonucleotide decreases UNC13A cryptic exon mRNA expression. In some embodiments, binding of the minimum region or a portion thereof to an antisense oligonucleotide increases UNC13A protein expression.

[0206] Table 3: Minimum Target Region of UNC13A mRNA

[0207]

[0208] In some cases, the antisense oligonucleotide comprises 13 to 30 nucleotides, wherein the antisense oligonucleotide is complementary to the sequence shown in any one of SEQ ID NOs: 1339 - 1342 or a portion thereof. In some embodiments, the antisense oligonucleotide is complementary to the sequence shown in SEQ ID NO: 1339 or a portion thereof. In some embodiments, the antisense oligonucleotide is complementary to the sequence shown in SEQ ID NO: 1340 or a portion thereof. In some embodiments, the antisense oligonucleotide is complementary to the sequence shown in SEQ ID NO: 1341 or a portion thereof. In some embodiments, the antisense oligonucleotide is complementary to the sequence shown in SEQ ID NO: 1342 or a portion thereof. In some embodiments, the antisense oligonucleotide is complementary to at least 1, at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, at least 11, at least 12, or at least 13 nucleotides of the sequence shown in any one of SEQ ID NOs: 1339 - 1342. In some embodiments, the antisense oligonucleotide is complementary to at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, at least 11, at least 12, or at least 13 consecutive nucleotides of the sequence shown in any one of SEQ ID NOs: 1339 - 1342. In some embodiments, the antisense oligonucleotide is complementary to 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, or 13 nucleotides of the sequence shown in any one of SEQ ID NOs: 1339 - 1342. In some embodiments, the antisense oligonucleotide is complementary to 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, or 13 consecutive nucleotides of the sequence shown in any one of SEQ ID NOs: 1339 - 1342. In some embodiments, the antisense oligonucleotide is at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99%, or 100% complementary to the sequence shown in SEQ ID NO: 1339. In some embodiments, the antisense oligonucleotide is at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99%, or 100% complementary to the sequence shown in SEQ ID NO: 1340. In some embodiments, the antisense oligonucleotide is at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99%, or 100% complementary to the sequence shown in SEQ ID NO: 1341. In some embodiments, the antisense oligonucleotide is at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99%, or 100% complementary to the sequence shown in SEQ ID NO: 1342.

[0209] In some embodiments, binding of the antisense oligonucleotide to the sequence or a portion thereof shown in any of SEQ ID NOs: 1339 - 1342 enhances UNC13A splicing regulatory activity. In some embodiments, binding of the antisense oligonucleotide to the sequence or a portion thereof shown in any of SEQ ID NOs: 1339 - 1342 increases UNC13A mRNA expression. In some embodiments, the expression of UNC13A mRNA in the cell is increased to at least 25%, at least 30%, at least 35%, at least 40%, at least 45% or at least 50% of the level of UNC13A mRNA expressed in wild - type cells. In some embodiments, the increase in the expression of UNC13A mRNA in the cell is from about 10% to about 95%, 20% to about 85%, 30% to about 75%, 40% to about 65% or 50% to about 55% of the level of UNC13A mRNA expressed in wild - type cells. In some embodiments, the expression of UNC13A mRNA in the cell is increased by at least 50%, at least 100%, at least 200%, at least 300%, at least 400% or at least 500%. In some embodiments, the increase in the expression of UNC13A mRNA in the cell is from about 25% to about 1000%, about 50% to about 800%, about 100% to about 700%, about 200% to about 600% or about 300% to about 500%. In some embodiments, binding of the antisense oligonucleotide to the sequence or a portion thereof shown in any of SEQ ID NOs: 1339 - 1342 reduces the expression of UNC13A cryptic exon mRNA. In some embodiments, the expression of UNC13A cryptic exon mRNA in the cell is reduced by at least 10%, at least 20%, at least 30%, at least 40% or at least 50%. In some embodiments, the expression of UNC13A cryptic exon mRNA in the cell is reduced from about 10% to about 90%, about 20% to about 80%, about 30% to about 70% or about 40% to about 60%. In some embodiments, the antisense oligonucleotide consists of the sequence shown in any of SEQ ID NOs: 1059 - 1074, 1075 - 1089, 1090 - 1103, 1248 - 1270, 1271 - 1292 and 1293 - 1313. In some embodiments, the antisense oligonucleotide consists of the sequence shown in any of SEQ ID NOs: 1059 - 1074, 1075 - 1089 and 1090 - 1103. In some embodiments, the antisense oligonucleotide consists of the sequence shown in any of SEQ ID NOs: 1248 - 1270, 1271 - 1292 and 1293 - 1313.

[0210] In some embodiments, the wild-type cells are cells from an individual who does not have amyotrophic lateral sclerosis (ALS) or frontotemporal dementia (FTD). In some embodiments, the wild-type cells are cells that express normal levels of the TDP-43 protein. In some embodiments, the wild-type cells are cells that do not produce transcripts of UNC13A that contain cryptic exons.

[0211] In some aspects, the present disclosure provides antisense oligonucleotides that comprise a sequence shown in any one of SEQ ID NOs: 20-355, 400-638, 810-959, or 961-1332. In some embodiments, the antisense oligonucleotide comprises or consists of 13 to 30 nucleotides. In some embodiments, the antisense oligonucleotide comprises or consists of 13 to 30, 15 to 28, 17 to 26, 19 to 24, or 21 to 23 nucleotides. In some embodiments, the antisense oligonucleotide comprises or consists of 13 to 25 nucleotides. In some embodiments, the antisense oligonucleotide comprises or consists of 13, 15, 17, 18, 19, 20, 21, 22, or 24 nucleotides. In some embodiments, the antisense oligonucleotide comprises or consists of 19, 20, 21, or 22 nucleotides. In some embodiments, the antisense oligonucleotide comprises or consists of at least 13, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, or 25 nucleotides. In some embodiments, the antisense oligonucleotide comprises or consists of no more than 25, 24, 23, 22, 21, 20, 19, 18, 17, 16, 15, or 13 nucleotides. In some embodiments, the antisense oligonucleotide comprises or consists of 13 nucleotides. In some embodiments, the antisense oligonucleotide comprises or consists of 15 nucleotides. In some embodiments, the antisense oligonucleotide comprises or consists of 17 nucleotides. In some embodiments, the antisense oligonucleotide comprises or consists of 18 nucleotides. In some embodiments, the antisense oligonucleotide comprises or consists of 19 nucleotides. In some embodiments, the antisense oligonucleotide comprises or consists of 20 nucleotides. In some embodiments, the antisense oligonucleotide comprises or consists of 21 nucleotides. In some embodiments, the antisense oligonucleotide comprises or consists of 22 nucleotides. In some embodiments, the antisense oligonucleotide comprises or consists of 23 nucleotides. In some embodiments, the antisense oligonucleotide comprises or consists of 24 nucleotides. In some embodiments, the antisense oligonucleotide comprises or consists of 25 nucleotides. In some embodiments, wherein the antisense oligonucleotide is provided to a cell, the expression of UNC13A mRNA in the cell is increased to at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, or at least 50% of the level of UNC13A mRNA expressed in wild-type cells. In some embodiments, the increase in the expression of UNC13A mRNA in the cell is about 10% to about 95%, 20% to about 85%, 30% to about 75%, 40% to about 65%, or 50% to about 55% of the level of UNC13A mRNA expressed in wild-type cells.In some embodiments, the expression of UNC13A mRNA in the cells is increased by about 25% to about 1000%, about 50% to about 800%, about 100% to about 700%, about 200% to about 600%, or about 300% to about 500%. In some embodiments, wherein an antisense oligonucleotide is provided to the cells, the expression of UNC13A cryptic exon mRNA in the cells is decreased by about 10% to about 90%, about 20% to about 80%, about 30% to about 70%, or about 40% to about 60%. In some embodiments, wherein an antisense oligonucleotide is provided to the cells, the expression of UNC13A protein in the cells is increased to at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, or at least 50% of the level of UNC13A protein expressed in wild-type cells. In some embodiments, the expression of UNC13A protein in the cells is increased by about 10% to about 95%, 20% to about 85%, 30% to about 75%, 40% to about 65%, or 50% to about 55% of the level of UNC13A protein expressed in wild-type cells. In some embodiments, the expression of UNC13A protein in the cells is increased by about 25% to about 1000%, about 50% to about 800%, about 100% to about 700%, about 200% to about 600%, or about 300% to about 500%. In some embodiments, wherein an antisense oligonucleotide is provided to the cells, the expression of UNC13A protein is increased to a level sufficient to improve the function of one or more neurons selected from the group consisting of: neuronal signaling, neuronal morphology, neurite length, neurotransmitter release, action potential, neuronal synaptic plasticity, and neuronal lifespan. In some embodiments, the antisense oligonucleotide consists of the sequence shown in any one of SEQ ID NO: 1059-1074, 1075-1089, 1090-1103, 1248-1270, 1271-1292, and 1293-1313. In some embodiments, the antisense oligonucleotide consists of the sequence shown in any one of SEQ ID NO: 1059-1074, 1075-1089, and 1090-1103. In some embodiments, the antisense oligonucleotide consists of the sequence shown in any one of SEQ ID NO: 1248-1270, 1271-1292, and 1293-1313. In some embodiments, the antisense oligonucleotide comprises a modification. In some embodiments, the modification is selected from the group consisting of: 2'OMe antisense oligonucleotide, 2'O-methoxyethyl antisense oligonucleotide, phosphorothioate antisense oligonucleotide, and locked nucleic acid (LNA) antisense oligonucleotide.

[0212] In some embodiments, the antisense oligonucleotide is complementary to the sequence shown in SEQ ID NO:1. In some embodiments, the antisense oligonucleotide comprises or consists of the sequence shown in any one of SEQ ID NOs: 20-355, 400-638, 810-959, or 961-1332. In some embodiments, the antisense oligonucleotide comprises or consists of 13 to 30 nucleotides. In some embodiments, the antisense oligonucleotide comprises a modification.

[0213] In some embodiments, the antisense oligonucleotide consists of the sequence shown in any one of SEQ ID NOs: 20-355, 400-638, 810-959, or 961-1332. In some embodiments, the antisense oligonucleotide comprises or consists of 13 to 30 nucleotides. In some embodiments, the antisense oligonucleotide comprises a modification.

[0214] In some embodiments, the antisense oligonucleotide comprises 13 nucleobases complementary to the sequence shown in SEQ ID NO:1, and wherein the antisense oligonucleotide comprises the sequence shown in any of SEQ ID NOs: 961 - 983 and 1116 - 1145. In some embodiments, the antisense oligonucleotide consists of 13 nucleotides complementary to the sequence shown in SEQ ID NO:1, and wherein the antisense oligonucleotide consists of the sequence shown in any of SEQ ID NOs: 961 - 983 and 1116 - 1145. In some embodiments, wherein the antisense oligonucleotide is provided to a cell, the expression of UNC13A mRNA in the cell is increased to at least 25%, at least 30%, at least 35%, at least 40%, at least 45% or at least 50% of the level of UNC13A mRNA expressed in wild - type cells. In some embodiments, the increase in the expression of UNC13A mRNA in the cell is from about 10% to about 95%, 20% to about 85%, 30% to about 75%, 40% to about 65% or 50% to about 55% of the level of UNC13A mRNA expressed in wild - type cells. In some embodiments, the increase in the expression of UNC13A mRNA in the cell is from about 25% to about 1000%, about 50% to about 800%, about 100% to about 700%, about 200% to about 600% or about 300% to about 500%. In some embodiments, wherein the antisense oligonucleotide is provided to a cell, the expression of UNC13A cryptic exon mRNA in the cell is decreased by about 10% to about 90%, about 20% to about 80%, about 30% to about 70% or about 40% to about 60%. In some embodiments, wherein the antisense oligonucleotide is provided to a cell, the expression of UNC13A protein in the cell is increased to at least 25%, at least 30%, at least 35%, at least 40%, at least 45% or at least 50% of the level of UNC13A protein expressed in wild - type cells. In some embodiments, the increase in the expression of UNC13A protein in the cell is from about 10% to about 95%, 20% to about 85%, 30% to about 75%, 40% to about 65% or 50% to about 55% of the level of UNC13A protein expressed in wild - type cells. In some embodiments, the increase in the expression of UNC13A protein in the cell is from about 25% to about 1000%, about 50% to about 800%, about 100% to about 700%, about 200% to about 600% or about 300% to about 500%. In some embodiments, wherein the antisense oligonucleotide is provided to a cell, the expression of UNC13A protein is increased to a level sufficient to improve the function of one or more neurons selected from the group consisting of: neuronal signaling, neuronal morphology, neurite length, neurotransmitter release, action potential, neuronal synaptic plasticity and neuronal lifespan.In some embodiments, the antisense oligonucleotide consists of the sequence shown in any one of SEQ ID NOs: 1059-1074, 1075-1089, 1090-1103, 1248-1270, 1271-1292, and 1293-1313. In some embodiments, the antisense oligonucleotide consists of the sequence shown in any one of SEQ ID NOs: 1059-1074, 1075-1089, and 1090-1103. In some embodiments, the antisense oligonucleotide consists of the sequence shown in any one of SEQ ID NOs: 1248-1270, 1271-1292, and 1293-1313. In some embodiments, the antisense oligonucleotide comprises a modification. In some embodiments, the modification is selected from the group consisting of: 2'-OMe antisense oligonucleotide, 2'-O-methoxyethyl antisense oligonucleotide, phosphorothioate antisense oligonucleotide, and locked nucleic acid (LNA) antisense oligonucleotide.

[0215] In some embodiments, the antisense oligonucleotide comprises 15 nucleotides complementary to the sequence shown in SEQ ID NO:1, and wherein the antisense oligonucleotide comprises the sequence shown in any of SEQ ID NOs: 984 - 1004 and 1146 - 1173. In some embodiments, the antisense oligonucleotide consists of 15 nucleotides complementary to the sequence shown in SEQ ID NO:1, and wherein the antisense oligonucleotide consists of the sequence shown in any of SEQ ID NOs: 984 - 1004 and 1146 - 1173. In some embodiments, wherein the antisense oligonucleotide is provided to a cell, the expression of UNC13A mRNA in the cell is increased to at least 25%, at least 30%, at least 35%, at least 40%, at least 45% or at least 50% of the level of UNC13A mRNA expressed in wild-type cells. In some embodiments, the increase in the expression of UNC13A mRNA in the cell is from about 10% to about 95%, 20% to about 85%, 30% to about 75%, 40% to about 65% or 50% to about 55% of the level of UNC13A mRNA expressed in wild-type cells. In some embodiments, the increase in the expression of UNC13A mRNA in the cell is about 25% to about 1000%, about 50% to about 800%, about 100% to about 700%, about 200% to about 600% or about 300% to about 500%. In some embodiments, wherein the antisense oligonucleotide is provided to a cell, the expression of UNC13A cryptic exon mRNA in the cell is decreased by about 10% to about 90%, about 20% to about 80%, about 30% to about 70% or about 40% to about 60%. In some embodiments, wherein the antisense oligonucleotide is provided to a cell, the expression of UNC13A protein in the cell is increased to at least 25%, at least 30%, at least 35%, at least 40%, at least 45% or at least 50% of the level of UNC13A protein expressed in wild-type cells. In some embodiments, the increase in the expression of UNC13A protein in the cell is from about 10% to about 95%, 20% to about 85%, 30% to about 75%, 40% to about 65% or 50% to about 55% of the level of UNC13A protein expressed in wild-type cells. In some embodiments, the increase in the expression of UNC13A protein in the cell is about 25% to about 1000%, about 50% to about 800%, about 100% to about 700%, about 200% to about 600% or about 300% to about 500%. In some embodiments, wherein the antisense oligonucleotide is provided to a cell, the expression of UNC13A protein is increased to a level sufficient to improve the function of one or more neurons selected from the group consisting of: neuronal signaling, neuronal morphology, neurite length, neurotransmitter release, action potential, neuronal synaptic plasticity and neuronal lifespan.In some embodiments, the antisense oligonucleotide consists of the sequence shown in any one of SEQ ID NOs: 1059-1074, 1075-1089, 1090-1103, 1248-1270, 1271-1292, and 1293-1313. In some embodiments, the antisense oligonucleotide consists of the sequence shown in any one of SEQ ID NOs: 1059-1074, 1075-1089, and 1090-1103. In some embodiments, the antisense oligonucleotide consists of the sequence shown in any one of SEQ ID NOs: 1248-1270, 1271-1292, and 1293-1313. In some embodiments, the antisense oligonucleotide comprises a modification. In some embodiments, the modification is selected from the group consisting of: 2'-OMe antisense oligonucleotide, 2'-O-methoxyethyl antisense oligonucleotide, phosphorothioate antisense oligonucleotide, and locked nucleic acid (LNA) antisense oligonucleotide.

[0216] In some embodiments, the antisense oligonucleotide comprises 16 nucleotides complementary to the sequence shown in SEQ ID NO:1, and wherein the antisense oligonucleotide comprises the sequence shown in any one of SEQ ID NO:41, 42, 56, 57, 71, 72, 86, 87, 101, 102, 116, 117, 131, 132, 146, 147, 161, 162, 176, 177, 191, 192, 206, 207, 221, 222, 236, 237, 251, 252, 266, 267, 281, 282, 296, 297, 311, 326, 341, 810, 811, 812, 813, 826, 827, 835, 838, 841, 842, 851, 852, 861, 864, 867, 868, 877, 878, 895, 900, 901, 910, 934, and 935. In some embodiments, the antisense oligonucleotide consists of 16 nucleotides complementary to the sequence shown in SEQ ID NO:1, and wherein the antisense oligonucleotide consists of the sequence shown in any one of SEQ ID NO:41, 42, 56, 57, 71, 72, 86, 87, 101, 102, 116, 117, 131, 132, 146, 147, 161, 162, 176, 177, 191, 192, 206, 207, 221, 222, 236, 237, 251, 252, 266, 267, 281, 282, 296, 297, 311, 326, 341, 810, 811, 812, 813, 826, 827, 835, 838, 841, 842, 851, 852, 861, 864, 867, 868, 877, 878, 895, 900, 901, 910, 934, and 935. In some embodiments, wherein the antisense oligonucleotide is provided to the cell, the expression of UNC13A mRNA in the cell is increased to at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, or at least 50% of the level of UNC13A mRNA expressed in wild-type cells. In some embodiments, the increase in the expression of UNC13A mRNA in the cell is from about 10% to about 95%, 20% to about 85%, 30% to about 75%, 40% to about 65%, or 50% to about 55% of the level of UNC13A mRNA expressed in wild-type cells. In some embodiments, the increase in the expression of UNC13A mRNA in the cell is about 25% to about 1000%, about 50% to about 800%, about 100% to about 700%, about 200% to about 600%, or about 300% to about 500%.In some embodiments, where an antisense oligonucleotide is provided to a cell, the expression of UNC13A cryptic exon mRNA in the cell is reduced by about 10% to about 90%, about 20% to about 80%, about 30% to about 70%, or about 40% to about 60%. In some embodiments, where an antisense oligonucleotide is provided to a cell, the expression of UNC13A protein in the cell is increased to at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, or at least 50% of the level of UNC13A protein expressed in wild-type cells. In some embodiments, the expression of UNC13A protein in the cell is increased by about 10% to about 95%, 20% to about 85%, 30% to about 75%, 40% to about 65%, or 50% to about 55% of the level of UNC13A protein expressed in wild-type cells. In some embodiments, the expression of UNC13A protein in the cell is increased by about 25% to about 1000%, about 50% to about 800%, about 100% to about 700%, about 200% to about 600%, or about 300% to about 500%. In some embodiments, where an antisense oligonucleotide is provided to a cell, the expression of UNC13A protein is increased to a level sufficient to improve the function of one or more neurons selected from the group consisting of: neuronal signaling, neuronal morphology, neurite length, neurotransmitter release, action potential, neuronal synaptic plasticity, and neuronal lifespan. In some embodiments, the antisense oligonucleotide consists of the sequence shown in any one of SEQ ID NO: 1059-1074, 1075-1089, 1090-1103, 1248-1270, 1271-1292, and 1293-1313. In some embodiments, the antisense oligonucleotide consists of the sequence shown in any one of SEQ ID NO: 1059-1074, 1075-1089, and 1090-1103. In some embodiments, the antisense oligonucleotide consists of the sequence shown in any one of SEQ ID NO: 1248-1270, 1271-1292, and 1293-1313. In some embodiments, the antisense oligonucleotide comprises a modification. In some embodiments, the modification is selected from the group consisting of: 2' OMe antisense oligonucleotide, 2'-O-methoxyethyl antisense oligonucleotide, phosphorothioate antisense oligonucleotide, and locked nucleic acid (LNA) antisense oligonucleotide.

[0217] In some embodiments, the antisense oligonucleotide comprises 17 nucleotides complementary to the sequence shown in SEQ ID NO:1, and wherein the antisense oligonucleotide comprises the sequence shown in any one of SEQ ID NO:43, 58, 73, 88, 103, 118, 133, 148, 163, 178, 193, 208, 223, 238, 253, 268, 283, 298, 312, 313, 327, 328, 342, 343, 814, 815, 816, 828, 829, 836, 839, 843, 844, 853, 854, 862, 865, 869, 870, 879, 880, 881, 891, 896, 897, 902, 903, 1005 - 1023, and 1174 - 1199. In some embodiments, the antisense oligonucleotide consists of 17 nucleotides complementary to the sequence shown in SEQ ID NO:1, and wherein the antisense oligonucleotide consists of the sequence shown in any one of SEQ ID NO:43, 58, 73, 88, 103, 118, 133, 148, 163, 178, 193, 208, 223, 238, 253, 268, 283, 298, 312, 313, 327, 328, 342, 343, 814, 815, 816, 828, 829, 836, 839, 843, 844, 853, 854, 862, 865, 869, 870, 879, 880, 881, 891, 896, 897, 902, 903, 1005 - 1023, and 1174 - 1199. In some embodiments, wherein the antisense oligonucleotide is provided to the cell, the expression of UNC13A mRNA in the cell is increased to at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, or at least 50% of the level of UNC13A mRNA expressed in wild - type cells. In some embodiments, the increase in the expression of UNC13A mRNA in the cell is from about 10% to about 95%, 20% to about 85%, 30% to about 75%, 40% to about 65%, or 50% to about 55% of the level of UNC13A mRNA expressed in wild - type cells. In some embodiments, the increase in the expression of UNC13A mRNA in the cell is about 25% to about 1000%, about 50% to about 800%, about 100% to about 700%, about 200% to about 600%, or about 300% to about 500%. In some embodiments, wherein the antisense oligonucleotide is provided to the cell, the expression of UNC13A cryptic exon mRNA in the cell is reduced by about 10% to about 90%, about 20% to about 80%, about 30% to about 70%, or about 40% to about 60%.In some embodiments, where an antisense oligonucleotide is provided to a cell, the expression of UNC13A protein in the cell is increased to at least 25%, at least 30%, at least 35%, at least 40%, at least 45% or at least 50% of the level of UNC13A protein expressed in wild-type cells. In some embodiments, the increase in the expression of UNC13A protein in the cell is from about 10% to about 95%, 20% to about 85%, 30% to about 75%, 40% to about 65% or 50% to about 55% of the level of UNC13A protein expressed in wild-type cells. In some embodiments, the increase in the expression of UNC13A protein in the cell is about 25% to about 1000%, about 50% to about 800%, about 100% to about 700%, about 200% to about 600% or about 300% to about 500%. In some embodiments, where an antisense oligonucleotide is provided to a cell, the expression of UNC13A protein is increased to a level sufficient to improve the function of one or more neurons selected from the group consisting of: neuronal signaling, neuronal morphology, neurite length, neurotransmitter release, action potential, neuronal synaptic plasticity and neuronal lifespan. In some embodiments, the antisense oligonucleotide consists of the sequence shown in any one of SEQ ID NO: 1059-1074, 1075-1089, 1090-1103, 1248-1270, 1271-1292 and 1293-1313. In some embodiments, the antisense oligonucleotide consists of the sequence shown in any one of SEQ ID NO: 1059-1074, 1075-1089 and 1090-1103. In some embodiments, the antisense oligonucleotide consists of the sequence shown in any one of SEQ ID NO: 1248-1270, 1271-1292 and 1293-1313. In some embodiments, the antisense oligonucleotide comprises a modification. In some embodiments, the modification is selected from the group consisting of: 2'OMe antisense oligonucleotide, 2'O-methoxyethyl antisense oligonucleotide, phosphorothioate antisense oligonucleotide and locked nucleic acid (LNA) antisense oligonucleotide.

[0218] In some embodiments, the antisense oligonucleotide comprises 18 nucleotides complementary to the sequence shown in SEQ ID NO:1, and wherein the antisense oligonucleotide comprises the sequence shown in any one of SEQ ID NOs: 38 - 40, 44, 45, 59, 60, 74, 75, 89, 90, 104, 105, 119, 120, 134, 135, 149, 150, 164, 165, 179, 180, 194, 195, 209, 210, 224, 225, 239, 240, 254, 255, 269, 270, 284, 285, 299, 300, 314, 329, 344, 518 - 638, 817, 818, 845, 855, 871, 882, 904, 958, 959, 960, 1024 - 1041, and 1200 - 1224. In some embodiments, the antisense oligonucleotide consists of 18 nucleotides complementary to the sequence shown in SEQ ID NO:1, and wherein the antisense oligonucleotide consists of the sequence shown in any one of SEQ ID NOs: 38 - 40, 44, 45, 59, 60, 74, 75, 89, 90, 104, 105, 119, 120, 134, 135, 149, 150, 164, 165, 179, 180, 194, 195, 209, 210, 224, 225, 239, 240, 254, 255, 269, 270, 284, 285, 299, 300, 314, 329, 344, 518 - 638, 817, 818, 845, 855, 871, 882, 904, 958, 959, 960, 1024 - 1041, and 1200 - 1224. In some embodiments, wherein an antisense oligonucleotide is provided to a cell, the expression of UNC13A mRNA in the cell is increased to at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, or at least 50% of the level of UNC13A mRNA expressed in wild - type cells. In some embodiments, the increase in the expression of UNC13A mRNA in the cell is from about 10% to about 95%, from 20% to about 85%, from 30% to about 75%, from 40% to about 65%, or from 50% to about 55% of the level of UNC13A mRNA expressed in wild - type cells. In some embodiments, the increase in the expression of UNC13A mRNA in the cell is from about 25% to about 1000%, from about 50% to about 800%, from about 100% to about 700%, from about 200% to about 600%, or from about 300% to about 500%. In some embodiments, wherein an antisense oligonucleotide is provided to a cell, the expression of UNC13A cryptic exon mRNA in the cell is reduced by about 10% to about 90%, about 20% to about 80%, about 30% to about 70%, or about 40% to about 60%.In some embodiments, where an antisense oligonucleotide is provided to a cell, the expression of UNC13A protein in the cell is increased to at least 25%, at least 30%, at least 35%, at least 40%, at least 45% or at least 50% of the level of UNC13A protein expressed in wild-type cells. In some embodiments, the increase in the expression of UNC13A protein in the cell is from about 10% to about 95%, 20% to about 85%, 30% to about 75%, 40% to about 65% or 50% to about 55% of the level of UNC13A protein expressed in wild-type cells. In some embodiments, the increase in the expression of UNC13A protein in the cell is about 25% to about 1000%, about 50% to about 800%, about 100% to about 700%, about 200% to about 600% or about 300% to about 500%. In some embodiments, where an antisense oligonucleotide is provided to a cell, the expression of UNC13A protein is increased to a level sufficient to improve the function of one or more neurons selected from the group consisting of: neuronal signaling, neuronal morphology, neurite length, neurotransmitter release, action potential, neuronal synaptic plasticity, and neuronal lifespan. In some embodiments, the antisense oligonucleotide consists of a sequence shown in any one of SEQ ID NOs: 1059-1074, 1075-1089, 1090-1103, 1248-1270, 1271-1292, and 1293-1313. In some embodiments, the antisense oligonucleotide consists of a sequence shown in any one of SEQ ID NOs: 1059-1074, 1075-1089, and 1090-1103. In some embodiments, the antisense oligonucleotide consists of a sequence shown in any one of SEQ ID NOs: 1248-1270, 1271-1292, and 1293-1313. In some embodiments, the antisense oligonucleotide comprises a modification. In some embodiments, the modification is selected from the group consisting of: 2'-OMe antisense oligonucleotide, 2'-O-methoxyethyl antisense oligonucleotide, phosphorothioate antisense oligonucleotide, and locked nucleic acid (LNA) antisense oligonucleotide.

[0219] In some embodiments, the antisense oligonucleotide comprises 19 nucleotides complementary to the sequence shown in SEQ ID NO:1, and wherein the antisense oligonucleotide comprises the sequence shown in any one of SEQ ID No:46, 61, 76, 91, 106, 121, 136, 151, 166, 181, 196, 211, 226, 241, 256, 271, 286, 301, 315, 316, 330, 331, 345, 346, 819, 830, 846, 856, 872, 883, 884, 892, 905, 1042 - 1058, and 1225 - 1247. In some embodiments, the antisense oligonucleotide consists of 19 nucleotides complementary to the sequence shown in SEQ ID NO:1, and wherein the antisense oligonucleotide consists of the sequence shown in any one of SEQ ID No:46, 61, 76, 91, 106, 121, 136, 151, 166, 181, 196, 211, 226, 241, 256, 271, 286, 301, 315, 316, 330, 331, 345, 346, 819, 830, 846, 856, 872, 883, 884, 892, 905, 1042 - 1058, and 1225 - 1247. In some embodiments, wherein the antisense oligonucleotide is provided to the cell, the expression of UNC13A mRNA in the cell is increased to at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, or at least 50% of the level of UNC13A mRNA expressed in wild-type cells. In some embodiments, the increase in the expression of UNC13A mRNA in the cell is about 10% to about 95%, 20% to about 85%, 30% to about 75%, 40% to about 65%, or 50% to about 55% of the level of UNC13A mRNA expressed in wild-type cells. In some embodiments, the increase in the expression of UNC13A mRNA in the cell is about 25% to about 1000%, about 50% to about 800%, about 100% to about 700%, about 200% to about 600%, or about 300% to about 500%. In some embodiments, wherein the antisense oligonucleotide is provided to the cell, the expression of UNC13A cryptic exon mRNA in the cell is decreased by about 10% to about 90%, about 20% to about 80%, about 30% to about 70%, or about 40% to about 60%. In some embodiments, wherein the antisense oligonucleotide is provided to the cell, the expression of UNC13A protein in the cell is increased to at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, or at least 50% of the level of UNC13A protein expressed in wild-type cells.In some embodiments, the increase in the expression of UNC13A protein in the cell is about 10% to about 95%, 20% to about 85%, 30% to about 75%, 40% to about 65%, or 50% to about 55% of the level of UNC13A protein expressed in wild-type cells. In some embodiments, the increase in the expression of UNC13A protein in the cell is about 25% to about 1000%, about 50% to about 800%, about 100% to about 700%, about 200% to about 600%, or about 300% to about 500%. In some embodiments, wherein an antisense oligonucleotide is provided to the cell, the expression of UNC13A protein is increased to a level sufficient to improve one or more neuronal functions selected from the group consisting of: neuronal signaling, neuronal morphology, neurite length, neurotransmitter release, action potential, neuronal synaptic plasticity, and neuronal lifespan. In some embodiments, the antisense oligonucleotide consists of the sequence shown in any one of SEQ ID NO: 1059-1074, 1075-1089, 1090-1103, 1248-1270, 1271-1292, and 1293-1313. In some embodiments, the antisense oligonucleotide consists of the sequence shown in any one of SEQ ID NO: 1059-1074, 1075-1089, and 1090-1103. In some embodiments, the antisense oligonucleotide consists of the sequence shown in any one of SEQ ID NO: 1248-1270, 1271-1292, and 1293-1313. In some embodiments, the antisense oligonucleotide comprises a modification. In some embodiments, the modification is selected from the group consisting of: 2'-OMe antisense oligonucleotide, 2'-O-methoxyethyl antisense oligonucleotide, phosphorothioate antisense oligonucleotide, and locked nucleic acid (LNA) antisense oligonucleotide.

[0220] In some embodiments, the antisense oligonucleotide comprises 20 nucleotides complementary to the sequence shown in SEQ ID NO:1, and wherein the antisense oligonucleotide comprises the sequence shown in any one of SEQ ID NO:47, 48, 62, 63, 77, 78, 92, 93, 107, 108, 122, 123, 137, 138, 152, 153, 167, 168, 182, 183, 197, 198, 212, 213, 227, 228, 242, 243, 257, 258, 272, 273, 287, 288, 302, 303, 317, 332, 347, 911, 936, 937, 1059 - 1074, and 1248 - 1270. In some embodiments, the antisense oligonucleotide consists of 20 nucleotides complementary to the sequence shown in SEQ ID NO:1, and wherein the antisense oligonucleotide consists of the sequence shown in any one of SEQ ID NO:47, 48, 62, 63, 77, 78, 92, 93, 107, 108, 122, 123, 137, 138, 152, 153, 167, 168, 182, 183, 197, 198, 212, 213, 227, 228, 242, 243, 257, 258, 272, 273, 287, 288, 302, 303, 317, 332, 347, 911, 936, 937, 1059 - 1074, and 1248 - 1270. In some embodiments, wherein the antisense oligonucleotide is provided to the cell, the expression of UNC13A mRNA in the cell is increased to at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, or at least 50% of the level of UNC13A mRNA expressed in wild - type cells. In some embodiments, the increase in the expression of UNC13A mRNA in the cell is from about 10% to about 95%, 20% to about 85%, 30% to about 75%, 40% to about 65%, or 50% to about 55% of the level of UNC13A mRNA expressed in wild - type cells. In some embodiments, the increase in the expression of UNC13A mRNA in the cell is about 25% to about 1000%, about 50% to about 800%, about 100% to about 700%, about 200% to about 600%, or about 300% to about 500%. In some embodiments, wherein the antisense oligonucleotide is provided to the cell, the expression of UNC13A cryptic exon mRNA in the cell is decreased by about 10% to about 90%, about 20% to about 80%, about 30% to about 70%, or about 40% to about 60%. In some embodiments, wherein the antisense oligonucleotide is provided to the cell, the expression of UNC13A protein in the cell is increased to at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, or at least 50% of the level of UNC13A protein expressed in wild - type cells.In some embodiments, the increase in the expression of UNC13A protein in the cells is about 10% to about 95%, 20% to about 85%, 30% to about 75%, 40% to about 65%, or 50% to about 55% of the level of UNC13A protein expressed in wild-type cells. In some embodiments, the increase in the expression of UNC13A protein in the cells is about 25% to about 1000%, about 50% to about 800%, about 100% to about 700%, about 200% to about 600%, or about 300% to about 500%. In some embodiments, wherein an antisense oligonucleotide is provided to the cells, the expression of UNC13A protein is increased to a level sufficient to improve one or more neuronal functions selected from the group consisting of: neuronal signaling, neuronal morphology, neurite length, neurotransmitter release, action potential, neuronal synaptic plasticity, and neuronal lifespan. In some embodiments, the antisense oligonucleotide consists of the sequence shown in any one of SEQ ID NO: 1059 - 1074, 1075 - 1089, 1090 - 1103, 1248 - 1270, 1271 - 1292, and 1293 - 1313. In some embodiments, the antisense oligonucleotide consists of the sequence shown in any one of SEQ ID NO: 1059 - 1074, 1075 - 1089, and 1090 - 1103. In some embodiments, the antisense oligonucleotide consists of the sequence shown in any one of SEQ ID NO: 1248 - 1270, 1271 - 1292, and 1293 - 1313. In some embodiments, the antisense oligonucleotide comprises a modification. In some embodiments, the modification is selected from the group consisting of: 2'-OMe antisense oligonucleotide, 2'-O-methoxyethyl antisense oligonucleotide, phosphorothioate antisense oligonucleotide, and locked nucleic acid (LNA) antisense oligonucleotide.

[0221] In some embodiments, the antisense oligonucleotide comprises 21 nucleotides complementary to the sequence shown in SEQ ID NO:1, and wherein the antisense oligonucleotide comprises the sequence shown in any one of SEQ ID NOs: 20-37, 49, 64, 79, 94, 109, 124, 139, 154, 169, 184, 199, 214, 229, 244, 259, 274, 289, 304, 318, 319, 333, 334, 348, 349, 400-517, 1075-1089, and 1271-1292. In some embodiments, the antisense oligonucleotide consists of 21 nucleotides complementary to the sequence shown in SEQ ID NO:1, and wherein the antisense oligonucleotide consists of the sequence shown in any one of SEQ ID NOs: 20-37, 49, 64, 79, 94, 109, 124, 139, 154, 169, 184, 199, 214, 229, 244, 259, 274, 289, 304, 318, 319, 333, 334, 348, 349, 400-517, 1075-1089, and 1271-1292. In some embodiments, wherein the antisense oligonucleotide is provided to the cell, the expression of UNC13A mRNA in the cell is increased to at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, or at least 50% of the level of UNC13A mRNA expressed in wild-type cells. In some embodiments, the increase in the expression of UNC13A mRNA in the cell is from about 10% to about 95%, 20% to about 85%, 30% to about 75%, 40% to about 65%, or 50% to about 55% of the level of UNC13A mRNA expressed in wild-type cells. In some embodiments, the increase in the expression of UNC13A mRNA in the cell is about 25% to about 1000%, about 50% to about 800%, about 100% to about 700%, about 200% to about 600%, or about 300% to about 500%. In some embodiments, wherein the antisense oligonucleotide is provided to the cell, the expression of UNC13A cryptic exon mRNA in the cell is reduced by about 10% to about 90%, about 20% to about 80%, about 30% to about 70%, or about 40% to about 60%. In some embodiments, wherein the antisense oligonucleotide is provided to the cell, the expression of UNC13A protein in the cell is increased to at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, or at least 50% of the level of UNC13A protein expressed in wild-type cells. In some embodiments, the increase in the expression of UNC13A protein in the cell is from about 10% to about 95%, 20% to about 85%, 30% to about 75%, 40% to about 65%, or 50% to about 55% of the level of UNC13A protein expressed in wild-type cells.In some embodiments, the expression of UNC13A protein in the cells is increased by about 25% to about 1000%, about 50% to about 800%, about 100% to about 700%, about 200% to about 600%, or about 300% to about 500%. In some embodiments, wherein an antisense oligonucleotide is provided to the cells, the expression of UNC13A protein is increased to a level sufficient to improve the function of one or more neurons selected from the group consisting of: neuronal signaling, neuronal morphology, neurite length, neurotransmitter release, action potential, neuronal synaptic plasticity, and neuronal lifespan. In some embodiments, the antisense oligonucleotide consists of the sequence shown in any one of SEQ ID NO: 1059-1074, 1075-1089, 1090-1103, 1248-1270, 1271-1292, and 1293-1313. In some embodiments, the antisense oligonucleotide consists of the sequence shown in any one of SEQ ID NO: 1059-1074, 1075-1089, and 1090-1103. In some embodiments, the antisense oligonucleotide consists of the sequence shown in any one of SEQ ID NO: 1248-1270, 1271-1292, and 1293-1313. In some embodiments, the antisense oligonucleotide comprises a modification. In some embodiments, the modification is selected from the group consisting of: 2' OMe antisense oligonucleotide, 2' O-methoxyethyl antisense oligonucleotide, phosphorothioate antisense oligonucleotide, and locked nucleic acid (LNA) antisense oligonucleotide.

[0222] In some embodiments, the antisense oligonucleotide comprises 22 nucleotides complementary to the sequence shown in SEQ ID NO:1, and wherein the antisense oligonucleotide comprises the sequence shown in any one of SEQ ID NO:50, 51, 65, 66, 80, 81, 95, 96, 110, 111, 125, 126, 140, 141, 155, 156, 170, 171, 185, 186, 200, 201, 215, 216, 230, 231, 245, 246, 260, 261, 275, 276, 290, 291, 305, 306, 320, 335, 350, 912, 913, 914, 938, 939, 940, 941, 1090 - 1103, and 1293 - 1313. In some embodiments, the antisense oligonucleotide consists of 22 nucleotides complementary to the sequence shown in SEQ ID NO:1, and wherein the antisense oligonucleotide consists of the sequence shown in any one of SEQ ID NO:50, 51, 65, 66, 80, 81, 95, 96, 110, 111, 125, 126, 140, 141, 155, 156, 170, 171, 185, 186, 200, 201, 215, 216, 230, 231, 245, 246, 260, 261, 275, 276, 290, 291, 305, 306, 320, 335, 350, 912, 913, 914, 938, 939, 940, 941, 1090 - 1103, and 1293 - 1313. In some embodiments, wherein the antisense oligonucleotide is provided to the cell, the expression of UNC13A mRNA in the cell is increased to at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, or at least 50% of the level of UNC13A mRNA expressed in wild - type cells. In some embodiments, the increase in the expression of UNC13A mRNA in the cell is from about 10% to about 95%, 20% to about 85%, 30% to about 75%, 40% to about 65%, or 50% to about 55% of the level of UNC13A mRNA expressed in wild - type cells. In some embodiments, the increase in the expression of UNC13A mRNA in the cell is about 25% to about 1000%, about 50% to about 800%, about 100% to about 700%, about 200% to about 600%, or about 300% to about 500%. In some embodiments, wherein the antisense oligonucleotide is provided to the cell, the expression of UNC13A cryptic exon mRNA in the cell is decreased by about 10% to about 90%, about 20% to about 80%, about 30% to about 70%, or about 40% to about 60%.In some embodiments, where an antisense oligonucleotide is provided to a cell, the expression of UNC13A protein in the cell is increased to at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, or at least 50% of the level of UNC13A protein expressed in wild-type cells. In some embodiments, the increase in the expression of UNC13A protein in the cell is from about 10% to about 95%, 20% to about 85%, 30% to about 75%, 40% to about 65%, or 50% to about 55% of the level of UNC13A protein expressed in wild-type cells. In some embodiments, the increase in the expression of UNC13A protein in the cell is about 25% to about 1000%, about 50% to about 800%, about 100% to about 700%, about 200% to about 600%, or about 300% to about 500%. In some embodiments, where an antisense oligonucleotide is provided to a cell, the expression of UNC13A protein is increased to a level sufficient to improve the function of one or more neurons selected from the group consisting of: neuronal signaling, neuronal morphology, neurite length, neurotransmitter release, action potential, neuronal synaptic plasticity, and neuronal lifespan. In some embodiments, the antisense oligonucleotide consists of the sequence shown in any one of SEQ ID NO: 1059 - 1074, 1075 - 1089, 1090 - 1103, 1248 - 1270, 1271 - 1292, and 1293 - 1313. In some embodiments, the antisense oligonucleotide consists of the sequence shown in any one of SEQ ID NO: 1059 - 1074, 1075 - 1089, and 1090 - 1103. In some embodiments, the antisense oligonucleotide consists of the sequence shown in any one of SEQ ID NO: 1248 - 1270, 1271 - 1292, and 1293 - 1313. In some embodiments, the antisense oligonucleotide comprises a modification. In some embodiments, the modification is selected from the group consisting of: 2'-OMe antisense oligonucleotide, 2'-O-methoxyethyl antisense oligonucleotide, phosphorothioate antisense oligonucleotide, and locked nucleic acid (LNA) antisense oligonucleotide.

[0223] In some embodiments, the antisense oligonucleotide comprises 23 nucleotides complementary to the sequence shown in SEQ ID NO:1, and wherein the antisense oligonucleotide comprises the sequence shown in any one of SEQ ID No:52, 67, 82, 97, 112, 127, 142, 157, 172, 187, 202, 217, 232, 247, 262, 277, 292, 307, 321, 322, 336, 337, 351, 352, 820, 831, 847, 857, 873, 885, 886, 893, 906, 915-918, 930 and 942-945. In some embodiments, the antisense oligonucleotide consists of 23 nucleotides complementary to the sequence shown in SEQ ID NO:1, and wherein the antisense oligonucleotide consists of the sequence shown in any one of SEQ ID No:52, 67, 82, 97, 112, 127, 142, 157, 172, 187, 202, 217, 232, 247, 262, 277, 292, 307, 321, 322, 336, 337, 351, 352, 820, 831, 847, 857, 873, 885, 886, 893, 906, 915-918, 930 and 942-945. In some embodiments, wherein the antisense oligonucleotide is provided to the cell, the expression of UNC13A mRNA in the cell is increased to at least 25%, at least 30%, at least 35%, at least 40%, at least 45% or at least 50% of the level of UNC13A mRNA expressed in wild-type cells. In some embodiments, the increase in the expression of UNC13A mRNA in the cell is about 10% to about 95%, 20% to about 85%, 30% to about 75%, 40% to about 65% or 50% to about 55% of the level of UNC13A mRNA expressed in wild-type cells. In some embodiments, the increase in the expression of UNC13A mRNA in the cell is about 25% to about 1000%, about 50% to about 800%, about 100% to about 700%, about 200% to about 600% or about 300% to about 500%. In some embodiments, wherein the antisense oligonucleotide is provided to the cell, the expression of UNC13A cryptic exon mRNA in the cell is decreased by about 10% to about 90%, about 20% to about 80%, about 30% to about 70% or about 40% to about 60%. In some embodiments, wherein the antisense oligonucleotide is provided to the cell, the expression of UNC13A protein in the cell is increased to at least 25%, at least 30%, at least 35%, at least 40%, at least 45% or at least 50% of the level of UNC13A protein expressed in wild-type cells.In some embodiments, the increased expression of UNC13A protein in the cell is about 10% to about 95%, 20% to about 85%, 30% to about 75%, 40% to about 65%, or 50% to about 55% of the level of UNC13A protein expressed in wild-type cells. In some embodiments, the increased expression of UNC13A protein in the cell is about 25% to about 1000%, about 50% to about 800%, about 100% to about 700%, about 200% to about 600%, or about 300% to about 500%. In some embodiments, wherein an antisense oligonucleotide is provided to the cell, the expression of UNC13A protein is increased to a level sufficient to improve one or more neuronal functions selected from the group consisting of: neuronal signaling, neuronal morphology, neurite length, neurotransmitter release, action potential, neuronal synaptic plasticity, and neuronal lifespan. In some embodiments, the antisense oligonucleotide consists of a sequence shown in any one of SEQ ID NO: 1059-1074, 1075-1089, 1090-1103, 1248-1270, 1271-1292, and 1293-1313. In some embodiments, the antisense oligonucleotide consists of a sequence shown in any one of SEQ ID NO: 1059-1074, 1075-1089, and 1090-1103. In some embodiments, the antisense oligonucleotide consists of a sequence shown in any one of SEQ ID NO: 1248-1270, 1271-1292, and 1293-1313. In some embodiments, the antisense oligonucleotide comprises a modification. In some embodiments, the modification is selected from the group consisting of: 2'-OMe antisense oligonucleotide, 2'-O-methoxyethyl antisense oligonucleotide, phosphorothioate antisense oligonucleotide, and locked nucleic acid (LNA) antisense oligonucleotide.

[0224] In some embodiments, the antisense oligonucleotide comprises 24 nucleotides complementary to the sequence shown in SEQ ID NO:1, and wherein the antisense oligonucleotide comprises the sequence shown in any one of SEQ ID NO:53, 54, 68, 69, 83, 84, 98, 99, 113, 114, 128, 129, 143, 144, 158, 159, 173, 174, 188, 189, 203, 204, 218, 219, 233, 234, 248, 249, 263, 264, 278, 279, 293, 294, 308, 309, 323, 338, 353, 821, 822, 832, 848, 858, 874, 887, 907, 919, 920 - 923, 931, 946 - 951, 1104 - 1115, and 1314 - 1332. In some embodiments, the antisense oligonucleotide consists of 24 nucleotides complementary to the sequence shown in SEQ ID NO:1, and wherein the antisense oligonucleotide consists of the sequence shown in any one of SEQ ID NO:53, 54, 68, 69, 83, 84, 98, 99, 113, 114, 128, 129, 143, 144, 158, 159, 173, 174, 188, 189, 203, 204, 218, 219, 233, 234, 248, 249, 263, 264, 278, 279, 293, 294, 308, 309, 323, 338, 353, 821, 822, 832, 848, 858, 874, 887, 907, 919, 920 - 923, 931, 946 - 951, 1104 - 1115, and 1314 - 1332. In some embodiments, wherein the antisense oligonucleotide is provided to a cell, the expression of UNC13A mRNA in the cell is increased to at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, or at least 50% of the level of UNC13A mRNA expressed in wild - type cells. In some embodiments, the increase in the expression of UNC13A mRNA in the cell is from about 10% to about 95%, 20% to about 85%, 30% to about 75%, 40% to about 65%, or 50% to about 55% of the level of UNC13A mRNA expressed in wild - type cells. In some embodiments, the increase in the expression of UNC13A mRNA in the cell is about 25% to about 1000%, about 50% to about 800%, about 100% to about 700%, about 200% to about 600%, or about 300% to about 500%. In some embodiments, wherein the antisense oligonucleotide is provided to a cell, the expression of UNC13A cryptic exon mRNA in the cell is reduced by about 10% to about 90%, about 20% to about 80%, about 30% to about 70%, or about 40% to about 60%.In some embodiments, where an antisense oligonucleotide is provided to a cell, the expression of UNC13A protein in the cell is increased to at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, or at least 50% of the level of UNC13A protein expressed in wild-type cells. In some embodiments, the increase in the expression of UNC13A protein in the cell is from about 10% to about 95%, 20% to about 85%, 30% to about 75%, 40% to about 65%, or 50% to about 55% of the level of UNC13A protein expressed in wild-type cells. In some embodiments, the increase in the expression of UNC13A protein in the cell is about 25% to about 1000%, about 50% to about 800%, about 100% to about 700%, about 200% to about 600%, or about 300% to about 500%. In some embodiments, where an antisense oligonucleotide is provided to a cell, the expression of UNC13A protein is increased to a level sufficient to improve the function of one or more neurons selected from the group consisting of: neuronal signaling, neuronal morphology, neurite length, neurotransmitter release, action potential, neuronal synaptic plasticity, and neuronal lifespan. In some embodiments, the antisense oligonucleotide consists of a sequence shown in any one of SEQ ID NO: 1059-1074, 1075-1089, 1090-1103, 1248-1270, 1271-1292, and 1293-1313. In some embodiments, the antisense oligonucleotide consists of a sequence shown in any one of SEQ ID NO: 1059-1074, 1075-1089, and 1090-1103. In some embodiments, the antisense oligonucleotide consists of a sequence shown in any one of SEQ ID NO: 1248-1270, 1271-1292, and 1293-1313. In some embodiments, the antisense oligonucleotide comprises a modification. In some embodiments, the modification is selected from the group consisting of: 2' OMe antisense oligonucleotide, 2' O-methoxyethyl antisense oligonucleotide, phosphorothioate antisense oligonucleotide, and locked nucleic acid (LNA) antisense oligonucleotide.

[0225] In some embodiments, the antisense oligonucleotide comprises 25 nucleotides complementary to the sequence shown in SEQ ID NO:1, and wherein the antisense oligonucleotide comprises the sequence shown in any one of SEQ ID No:55, 70, 85, 100, 115, 130, 145, 160, 175, 190, 205, 220, 235, 250, 265, 280, 295, 310, 324, 325, 339, 340, 354, 355, 823, 824, 825, 833, 834, 837, 840, 849, 850, 859, 860, 863, 866, 875, 876, 888 - 890, 894, 898, 899, 908, 909, 924 - 929, 932, 933, 952 and 953 - 957. In some embodiments, the antisense oligonucleotide consists of 25 nucleotides complementary to the sequence shown in SEQ ID NO:1, and wherein the antisense oligonucleotide consists of the sequence shown in any one of SEQ ID No:55, 70, 85, 100, 115, 130, 145, 160, 175, 190, 205, 220, 235, 250, 265, 280, 295, 310, 324, 325, 339, 340, 354, 355, 823, 824, 825, 833, 834, 837, 840, 849, 850, 859, 860, 863, 866, 875, 876, 888 - 890, 894, 898, 899, 908, 909, 924 - 929, 932, 933, 952 and 953 - 957. In some embodiments, wherein the antisense oligonucleotide is provided to the cell, the expression of UNC13A mRNA in the cell is increased to at least 25%, at least 30%, at least 35%, at least 40%, at least 45% or at least 50% of the level of UNC13A mRNA expressed in wild - type cells. In some embodiments, the increase in the expression of UNC13A mRNA in the cell is from about 10% to about 95%, 20% to about 85%, 30% to about 75%, 40% to about 65% or 50% to about 55% of the level of UNC13A mRNA expressed in wild - type cells. In some embodiments, the increase in the expression of UNC13A mRNA in the cell is about 25% to about 1000%, about 50% to about 800%, about 100% to about 700%, about 200% to about 600% or about 300% to about 500%. In some embodiments, wherein the antisense oligonucleotide is provided to the cell, the expression of UNC13A cryptic exon mRNA in the cell is decreased by about 10% to about 90%, about 20% to about 80%, about 30% to about 70% or about 40% to about 60%.In some embodiments, where an antisense oligonucleotide is provided to a cell, the expression of UNC13A protein in the cell is increased to at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, or at least 50% of the level of UNC13A protein expressed in wild-type cells. In some embodiments, the increase in the expression of UNC13A protein in the cell is from about 10% to about 95%, 20% to about 85%, 30% to about 75%, 40% to about 65%, or 50% to about 55% of the level of UNC13A protein expressed in wild-type cells. In some embodiments, the increase in the expression of UNC13A protein in the cell is about 25% to about 1000%, about 50% to about 800%, about 100% to about 700%, about 200% to about 600%, or about 300% to about 500%. In some embodiments, where an antisense oligonucleotide is provided to a cell, the expression of UNC13A protein is increased to a level sufficient to improve the function of one or more neurons selected from the group consisting of: neuronal signaling, neuronal morphology, neurite length, neurotransmitter release, action potential, neuronal synaptic plasticity, and neuronal lifespan. In some embodiments, the antisense oligonucleotide consists of the sequence shown in any one of SEQ ID NO: 1059 - 1074, 1075 - 1089, 1090 - 1103, 1248 - 1270, 1271 - 1292, and 1293 - 1313. In some embodiments, the antisense oligonucleotide consists of the sequence shown in any one of SEQ ID NO: 1059 - 1074, 1075 - 1089, and 1090 - 1103. In some embodiments, the antisense oligonucleotide consists of the sequence shown in any one of SEQ ID NO: 1248 - 1270, 1271 - 1292, and 1293 - 1313. In some embodiments, the antisense oligonucleotide comprises a modification. In some embodiments, the modification is selected from the group consisting of: 2'-OMe antisense oligonucleotide, 2'-O-methoxyethyl antisense oligonucleotide, phosphorothioate antisense oligonucleotide, and locked nucleic acid (LNA) antisense oligonucleotide.

[0226] In some embodiments, the antisense oligonucleotide comprises a modification. In some embodiments, the 5'-end and / or 3'-end of the antisense oligonucleotide comprises a modification. In some embodiments, the 5'-end or 3'-end of the antisense oligonucleotide comprises a modification. In some embodiments, the 5'-end and 3'-end of the antisense oligonucleotide comprise a modification. In some embodiments, the modification is selected from the group consisting of: 2'-OMe antisense oligonucleotide, 2'-O-methoxyethyl antisense oligonucleotide, phosphorothioate antisense oligonucleotide, and LNA antisense oligonucleotide. In some embodiments, the modification is a 2'-OMe antisense oligonucleotide.

[0227] In some embodiments, the 5'-end of the antisense oligonucleotide comprises a modification, and 1, 2, 3, 4, or 5 nucleotides are modified. In some embodiments, the 5'-end of the antisense oligonucleotide comprises a modification, and at least 1, at least 2, at least 3, at least 4, or at least 5 nucleotides are modified. In some embodiments, the 3'-end of the antisense oligonucleotide comprises a modification, and 1, 2, 3, 4, or 5 nucleotides are modified. In some embodiments, the 3'-end of the antisense oligonucleotide comprises a modification, and at least 1, at least 2, at least 3, at least 4, or at least 5 nucleotides are modified. In some embodiments, the 5'-end and 3'-end of the antisense oligonucleotide comprise a modification, and 1, 2, 3, 4, or 5 nucleotides are modified at each of the 5'-end and 3'-end of the antisense oligonucleotide. In some embodiments, the 5'-end and 3'-end of the antisense oligonucleotide comprise a modification, and at least 1, at least 2, at least 3, at least 4, or at least 5 nucleotides are modified at each of the 5'-end and 3'-end of the antisense oligonucleotide. In some embodiments, the modification is selected from the group consisting of: 2'-OMe antisense oligonucleotide, 2'-O-methoxyethyl antisense oligonucleotide, phosphorothioate antisense oligonucleotide, and LNA antisense oligonucleotide. In some embodiments, the modification is a 2'-OMe antisense oligonucleotide.

[0228] In some embodiments, the antisense oligonucleotide comprises a modification, and at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 95%, or at least 98% of the nucleotides comprise a modification. In some embodiments, 30% to 100%, 40% to 90%, 50% to 80%, or 60% to 70% of the nucleotides comprise a modification. In some embodiments, about 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 98%, 99%, or 100% of the nucleotides comprise a modification. In some embodiments, the modification is selected from the group consisting of: 2'-OMe antisense oligonucleotide, 2'-O-methoxyethyl antisense oligonucleotide, phosphorothioate antisense oligonucleotide, and LNA antisense oligonucleotide. In some embodiments, the modification is a 2'-OMe antisense oligonucleotide.

[0229] In some embodiments, the antisense oligonucleotide contains one or more modified nucleotides, such as nucleotides other than ribonucleotides (2'-hydroxy nucleotides), including but not limited to deoxynucleotides, nucleotide analogs, abasic nucleotides, 2'-modified nucleotides, 3'-to-3' linked (inverted) nucleotides, non-natural base-containing nucleotides, bridged nucleotides, peptide nucleic acids, 2',3'-dideoxy nucleotide analogs, locked nucleotides, 3'-O-methoxy nucleotides, 2'-F-arabinonucleotides, morpholino nucleotides, vinyl phosphonate deoxyribonucleotides and vinyl phosphonate nucleotides, 2'-O-methyl nucleotides, 2'-deoxy-2'-fluoro nucleotides, 2'-deoxy nucleotides, 2'-methoxyethyl nucleotides, 2'-amino nucleotides and 2'-alkyl nucleotides.

[0230] In embodiments, the antisense oligonucleotide is a modified antisense oligonucleotide. In embodiments, the modified antisense oligonucleotide comprises phosphoramidate morpholino oligonucleotides, phosphorodiamidate morpholino oligonucleotides, thiophosphate-modified oligonucleotides, 2'O-methyl (2'O-Me)-modified oligonucleotides, peptide nucleic acids (PNA), locked nucleic acids (LNA), dithiophosphate oligonucleotides, 2'O-methoxyethyl (2'-MOE)-modified oligonucleotides, 2'-fluoro-modified oligonucleotides, 2'O,4'C-ethylene-bridged nucleic acids (ENA), tricyclic-DNA, tricyclic-DNA thiophosphate nucleotides, constrained ethyl-bridged nucleic acids, 2'-O-[2-(N-methylcarbamoyl)ethyl]-modified oligonucleotides, morpholino oligonucleotides and peptide-conjugated phosphoramidate morpholino oligonucleotides (PPMO) or any combination thereof. In some embodiments, the modification of the antisense oligonucleotide increases the stability of the oligonucleotide, reduces the efficacy of inclusion of the cryptic exon or reduces the efficiency of inclusion of the cryptic exon. In some embodiments, the modification of the antisense oligonucleotide improves the delivery of the oligonucleotide to the UNC13 mRNA molecule.

[0231] In some embodiments, the ASO contains modifications that enhance the pharmacokinetic or biodistribution properties of the ASO trigger or the conjugate to which it is attached to improve the cell-specific or tissue-specific distribution and cell-specific uptake of the conjugate. In some cases, the binding of a moiety to a cell or cell receptor can initiate endocytosis. The modification can be monovalent, divalent, trivalent, tetravalent or of higher valency. Representative targeting moieties include but are not limited to compounds having an affinity for cell surface molecules, cell receptor ligands, haptens, antibodies, monoclonal antibodies, antibody fragments and antibody mimetics having an affinity for cell surface molecules.

[0232] In some embodiments, modification includes modifying ribonucleic acid (RNA) into deoxyribonucleic acid (DNA), threose nucleic acid (TNA), glycol nucleic acid (GNA), peptide nucleic acid (PNA), locked nucleic acid (LNA), or hybrids thereof.

[0233] In some embodiments, the ASO includes a modified sugar moiety. In some embodiments, the ASO includes a sugar substitute. In certain such embodiments, the oxygen atom of the sugar moiety is replaced by, for example, a sulfur, carbon, or nitrogen atom. In certain such embodiments, such modified sugar moieties further include bridging and / or non-bridging substituents. In some embodiments, morpholino is modified by adding or changing various substituent groups from the above morpholino structure.

[0234] In some embodiments, the ASO includes modified nucleoside variants that maintain proper base pairing. In some embodiments, the ASO includes modified A, modified C, modified G, or modified U. In one embodiment, the modified ASO includes modified C, such as 5-methylcytosine or 5-hydroxymethylcytosine; modified U, such as 5-methyluridine or replaced with thymine; or modified A, such as N6-methyladenine. In one embodiment, the ASO can include a mixture of unmodified nucleosides and modified nucleosides.

[0235] In some embodiments, the ASO includes a modified internucleoside linkage. In some embodiments, the backbone phosphate groups are modified by replacing one or more oxygen atoms with different substituents. In some embodiments, the backbone is formed by: mRNA (alternating phosphates and riboses), LNA (locked nucleic acid), tcDNA (tricyclic DNA), cEt (constrained ethyl-bridged nucleic acid); ENA (ethyl-bridged nucleic acid), HNA (hexitol nucleic acid), TNA (threose nucleic acid), PMO (phosphorodiamidate morpholino oligomer) PMO, PNA (peptide nucleic acid), 2'-OMe-RNA, 2'-O-methoxyethyl (MOE) nucleic acid, or 2-O-(2-methylcarbamoyl (MCE) nucleotides, or any combination thereof.

[0236] In some embodiments, the antisense oligonucleotides are used in a method of treatment or in the preparation of a medicament for treatment, wherein the antisense oligonucleotides are administered to a patient in need of increased UNC13A mRNA expression. In some embodiments, the expression of UNC13A mRNA in the cells is increased by at least 50%, at least 100%, at least 200%, at least 300%, at least 400% or at least 500%. In some embodiments, the expression of UNC13A mRNA in the cells is increased by about 25% to about 1000%, about 50% to about 800%, about 100% to about 700%, about 200% to about 600% or about 300% to about 500%. In some embodiments, the expression of UNC13A mRNA in the cells is increased by about 100%, about 200%, about 300%, about 400% or about 500%. In some embodiments, the antisense oligonucleotides increase the expression of UNC13A protein in the cells to at least 25%, at least 30%, at least 35%, at least 40%, at least 45% or at least 50% of the level of UNC13A mRNA expressed in wild-type cells. In some embodiments, the expression of UNC13A mRNA in the cells is increased by about 10% to about 95%, 20% to about 85%, 30% to about 75%, 40% to about 65% or 50% to about 55% of the level of UNC13A mRNA expressed in wild-type cells. In some embodiments, the antisense oligonucleotides comprise or consist of the sequences shown in any one of SEQ ID NO: 20-355, 400-638, 810-959 or 961-1332. In some embodiments, the antisense oligonucleotides consist of the sequences shown in any one of SEQ ID NO: 1059-1074, 1075-1089, 1090-1103, 1248-1270, 1271-1292 and 1293-1313. In some embodiments, the antisense oligonucleotides consist of the sequences shown in any one of SEQ ID NO: 1059-1074, 1075-1089 and 1090-1103. In some embodiments, the antisense oligonucleotides consist of the sequences shown in any one of SEQ ID NO: 1248-1270, 1271-1292 and 1293-1313.

[0237] In some embodiments, the antisense oligonucleotide restores the expression of UNC13A mRNA in the cell to at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, or at least 50% of the level of UNC13A mRNA expressed in wild-type cells. In some embodiments, the expression of UNC13A protein in the cell is restored to about 10% to about 95%, 20% to about 85%, 30% to about 75%, 40% to about 65%, or 50% to about 55% of the level of UNC13A protein expressed in wild-type cells. In some embodiments, the antisense oligonucleotide comprises or consists of the sequence shown in any one of SEQ ID NO: 20 - 355, 400 - 638, 810 - 959, or 961 - 1332. In some embodiments, the antisense oligonucleotide consists of the sequence shown in any one of SEQ ID NO: 1059 - 1074, 1075 - 1089, 1090 - 1103, 1248 - 1270, 1271 - 1292, and 1293 - 1313. In some embodiments, the antisense oligonucleotide consists of the sequence shown in any one of SEQ ID NO: 1059 - 1074, 1075 - 1089, and 1090 - 1103. In some embodiments, the antisense oligonucleotide consists of the sequence shown in any one of SEQ ID NO: 1248 - 1270, 1271 - 1292, and 1293 - 1313.

[0238] In some embodiments, the antisense oligonucleotides are used in a method of treatment or in the preparation of a medicament for treatment, wherein the antisense oligonucleotides are administered to a patient in need of reducing the expression of UNC13A cryptic exon mRNA. In some embodiments, the expression of UNC13A cryptic exon mRNA in a cell is reduced by at least 10%, at least 20%, at least 30%, at least 40% or at least 50%. In some embodiments, the expression of UNC13A cryptic exon mRNA in a cell is reduced by about 10% to about 90%, about 20% to about 80%, about 30% to about 70% or about 40% to about 60%. In some embodiments, the expression of UNC13A cryptic exon mRNA in a cell is reduced by about 10%, about 20%, about 30%, about 40% or about 50%. In some embodiments, the antisense oligonucleotide comprises or consists of the sequence shown in any one of SEQ ID NO: 20-355, 400-638, 810-959 or 961-1332. In some embodiments, the antisense oligonucleotide consists of the sequence shown in any one of SEQ ID NO: 1059-1074, 1075-1089, 1090-1103, 1248-1270, 1271-1292 and 1293-1313. In some embodiments, the antisense oligonucleotide consists of the sequence shown in any one of SEQ ID NO: 1059-1074, 1075-1089 and 1090-1103. In some embodiments, the antisense oligonucleotide consists of the sequence shown in any one of SEQ ID NO: 1248-1270, 1271-1292 and 1293-1313.

[0239] In some embodiments, the antisense oligonucleotide reduces the expression of the UNC13A cryptic exon mRNA by at least 10%, at least 20%, at least 30%, at least 40%, or at least 50% of the level of the UNC13A cryptic exon mRNA expressed in cells treated with shRNA targeting TARDBP. In some embodiments, the antisense oligonucleotide comprises or consists of the sequence shown in any one of SEQ ID NOs: 20-355, 400-638, 810-959, or 961-1332. In some embodiments, the antisense oligonucleotide consists of the sequence shown in any one of SEQ ID NOs: 1059-1074, 1075-1089, 1090-1103, 1248-1270, 1271-1292, and 1293-1313. In some embodiments, the antisense oligonucleotide consists of the sequence shown in any one of SEQ ID NOs: 1059-1074, 1075-1089, and 1090-1103. In some embodiments, the antisense oligonucleotide consists of the sequence shown in any one of SEQ ID NOs: 1248-1270, 1271-1292, and 1293-1313.

[0240] In some embodiments, the antisense oligonucleotides are used in a method of treatment or in the preparation of a medicament for treatment, wherein the antisense oligonucleotides are administered to a patient in need of increasing the expression of the UNC13A protein. In some embodiments, the expression of the UNC13A protein in the cell is increased by at least 50%, at least 100%, at least 200%, at least 300%, at least 400% or at least 500%. In some embodiments, the expression of the UNC13A protein in the cell is increased by about 25% to about 1000%, about 50% to about 800%, about 100% to about 700%, about 200% to about 600% or about 300% to about 500%. In some embodiments, the expression of the UNC13A protein in the cell is increased by about 100%, about 200%, about 300%, about 400% or about 500%. In some embodiments, the antisense oligonucleotide comprises or consists of the sequence shown in any one of SEQ ID NOs: 20-355, 400-638, 810-959 or 961-1332. In some embodiments, the antisense oligonucleotide consists of the sequence shown in any one of SEQ ID NOs: 1059-1074, 1075-1089, 1090-1103, 1248-1270, 1271-1292 and 1293-1313. In some embodiments, the antisense oligonucleotide consists of the sequence shown in any one of SEQ ID NOs: 1059-1074, 1075-1089 and 1090-1103. In some embodiments, the antisense oligonucleotide consists of the sequence shown in any one of SEQ ID NOs: 1248-1270, 1271-1292 and 1293-1313.

[0241] In some embodiments, the antisense oligonucleotides are used in a method of treatment or in the preparation of a medicament for treatment, wherein the antisense oligonucleotides are administered to a patient in need of increasing the expression of UNC13A protein to a level sufficient to improve one or more neuronal functions. In some embodiments, the one or more neuronal functions are selected from the group consisting of: neuronal signaling, neuronal morphology, neurite length, neurotransmitter release, action potential, neuronal synaptic plasticity, and neuronal lifespan. In some embodiments, the antisense oligonucleotides improve 1, 2, 3, 4, 5, or 6 neuronal functions selected from the group consisting of: neuronal signaling, neuronal morphology, neurite length, neurotransmitter release, action potential, neuronal synaptic plasticity, and neuronal lifespan. In some embodiments, the antisense oligonucleotides improve at least 1, at least 2, at least 3, at least 4, or at least 5 neuronal functions selected from the group consisting of: neuronal signaling, neuronal morphology, neurite length, neurotransmitter release, action potential, neuronal synaptic plasticity, and neuronal lifespan. In some embodiments, the antisense oligonucleotide comprises or consists of the sequence shown in any one of SEQ ID NO: 20-355, 400-638, 810-959, or 961-1332. In some embodiments, the antisense oligonucleotide consists of the sequence shown in any one of SEQ ID NO: 1059-1074, 1075-1089, 1090-1103, 1248-1270, 1271-1292, and 1293-1313. In some embodiments, the antisense oligonucleotide consists of the sequence shown in any one of SEQ ID NO: 1059-1074, 1075-1089, and 1090-1103. In some embodiments, the antisense oligonucleotide consists of the sequence shown in any one of SEQ ID NO: 1248-1270, 1271-1292, and 1293-1313.

[0242] In some embodiments, the antisense oligonucleotide restores the expression of UNC13A protein in the cell to at least 10%, at least 20%, at least 30%, at least 40%, or at least 50% of the level of UNC13A protein expressed in wild-type cells. In some embodiments, the antisense oligonucleotide comprises or consists of the sequence shown in any one of SEQ ID NO: 20-355, 400-638, 810-959, or 961-1332. In some embodiments, the antisense oligonucleotide consists of the sequence shown in any one of SEQ ID NO: 1059-1074, 1075-1089, 1090-1103, 1248-1270, 1271-1292, and 1293-1313. In some embodiments, the antisense oligonucleotide consists of the sequence shown in any one of SEQ ID NO: 1059-1074, 1075-1089, and 1090-1103. In some embodiments, the antisense oligonucleotide consists of the sequence shown in any one of SEQ ID NO: 1248-1270, 1271-1292, and 1293-1313.

[0243] In some embodiments, the antisense oligonucleotide increases UNC13A mRNA expression at about 4 days, about 8 days, about 12 days, about 16 days, about 20 days, or about 24 days after the cell contacts the ASO. In some embodiments, the antisense oligonucleotide is provided to the cell in multiple doses. In some embodiments, the antisense oligonucleotide increases UNC13A mRNA expression at about 4 days, about 8 days, about 12 days, about 16 days, about 20 days, or about 24 days after administering the ASO. In some embodiments, the antisense oligonucleotide increases UNC13A mRNA expression such that UNC13A expression is fully restored after administering the ASO.

[0244] In some embodiments, the antisense oligonucleotide decreases UNC13A cryptic exon mRNA expression at about 4 days, about 8 days, about 12 days, about 16 days, about 20 days, or about 24 days after the cell contacts the ASO. In some embodiments, the antisense oligonucleotide is provided to the cell in multiple doses. In some embodiments, the antisense oligonucleotide decreases UNC13A cryptic exon mRNA expression at about 4 days, about 8 days, about 12 days, about 16 days, about 20 days, or about 24 days after administering the ASO. In some embodiments, the antisense oligonucleotide reduces UNC13A cryptic exon mRNA expression such that UNC13A cryptic exon expression is fully reduced after administering the ASO.

[0245] In some embodiments, the antisense oligonucleotide increases UNC13A protein expression at about 4 days, about 8 days, about 12 days, about 16 days, about 20 days, about 24 days after the cell contacts the ASO. In some embodiments, the antisense oligonucleotide is provided to the cell in multiple doses. In some embodiments, the antisense oligonucleotide increases UNC13A protein expression at about 4 days, about 8 days, about 12 days, about 16 days, about 20 days, about 24 days after administering the ASO. In some embodiments, the antisense oligonucleotide increases UNC13A protein expression such that the increase in UNC13A protein expression is fully restored after administering the ASO.

[0246] In some embodiments, an antisense oligonucleotide is provided to a cell, wherein the cell is a human cell or a murine cell. In some embodiments, the cell is a human cell. In some embodiments, the cell is a murine cell. In some embodiments, an antisense oligonucleotide is provided to a cell, wherein the cell is a neuron. In some embodiments, the cell is a human neuron. In some embodiments, the cell is a murine neuron. In some embodiments, the neuron is a motor neuron, a cortical neuron, or an excitatory neuron. In some embodiments, the cell is heterozygous or homozygous for a risk allele. In some embodiments, the risk allele is selected from the group consisting of rs12608932 (hg38 chr19:17,641,880A→C), rs12973192 (hg38 chr19:17,642,430C→G), rs56041637 (hg38 chr19:17,642,033-17,642,056 0-2CATC repeat→3-5CATC repeat), and rs62121687 (hg38 chr19:17,642,351C→A) or any combination thereof. In some embodiments, the cell contains a mutation associated with amyotrophic lateral sclerosis (ALS) or frontotemporal dementia (FTD). In some embodiments, the antisense oligonucleotide comprises or consists of the sequence shown in any one of SEQ ID NOs: 20-355, 400-638, 810-959, or 961-1332.

[0247] In some embodiments, an antisense oligonucleotide is provided to a cell, wherein the cell is a human cell. In some embodiments, the cell comprises a mutation associated with amyotrophic lateral sclerosis (ALS) or frontotemporal dementia (FTD). In some embodiments, the mutation is a mutation in the TDP-43 protein. In some embodiments, the mutation in the TDP-43 protein is G298S. In some embodiments, the mutation in the TDP-43 protein is A382T. In some embodiments, the cell comprises a single nucleotide polymorphism (SNP). In some embodiments, the SNP is associated with the UNC13A protein. In some embodiments, the SNP is a risk allele, wherein the cell is heterozygous or homozygous for the risk allele. In some embodiments, the risk allele is selected from the group consisting of rs12608932 (hg38chr19:17.641,880A→C), rs12973192 (hg38chr19:17,642,430C→G), rs56041637 (hg38chr19:17,642,033-17,642,056 0-2CATC repeat→3-5CATC repeat), and rs62121687 (hg38chr19:17,642,351C→A) or any combination thereof. In some embodiments, the risk allele is rs12973192 (hg38 chr19:17,642,430C→G). In some embodiments, an antisense oligonucleotide is provided to a cell comprising an SNP, wherein the SNP is rs12973192 (hg38 chr19:17,642,430C→G). In some embodiments, the cell is selected from the group consisting of GM25256 (GM or GM cell line), NDS00262 (005 or 005 cell line), NDS00209 (4048 or 4048 cell line), and NDS00235 (4157 or 4157 cell line). In some embodiments, the cell is a healthy cell selected from the group consisting of GM25256 (GM or GM cell line) and NDS00262 (005 or 005 cell line). In some embodiments, the cell is a cell comprising a mutation in the TDP-43 protein selected from the group consisting of NDS00209 (4048 or 4048 cell line) and NDS00235 (4157 or 4157 cell line). In some embodiments, the cell is NDS00209 (4048 or 4048 cell line), wherein the mutation in the TDP-43 protein is G298S. In some embodiments, the cell is NDS00235 (4157 or 4157 cell line), wherein the mutation in the TDP-43 protein is A382T.In some embodiments, the antisense oligonucleotide comprises or consists of the sequence shown in any one of SEQ ID NOs: 20-355, 400-638, 810-959 or 961-1332.

[0248] In some embodiments, the antisense oligonucleotide is active in one or more iPSC neurons. In some embodiments, the antisense oligonucleotide is active in multiple iPSC neurons having different genetic backgrounds, disease mutations, or UNC13A SNP statuses. In some embodiments, the antisense oligonucleotide is active in multiple iPSC neurons having different genetic backgrounds, disease mutations, and UNC13A SNP statuses. In some embodiments, the antisense oligonucleotide comprises or consists of the sequence shown in any one of SEQ ID NOs: 20-355, 400-638, 810-959 or 961-1332.

[0249] In some embodiments, the antisense oligonucleotide is provided to a cell that expresses a low level of TDP-43 protein. In some embodiments, the cell expresses less than 50%, less than 45%, less than 40%, less than 35%, less than 30%, less than 25%, less than 20%, less than 15%, less than 10%, less than 5%, or less than 1% TDP-43 protein. In some embodiments, the antisense oligonucleotide comprises or consists of the sequence shown in any one of SEQ ID NOs: 20-355, 400-638, 810-959 or 961-1332.

[0250] In some embodiments, the antisense oligonucleotide is provided to the cell by transfection. In some embodiments, the antisense oligonucleotide is provided to the cell by other delivery methods known in the art.

[0251] In some embodiments, the antisense oligonucleotide does not cause any cell morphological changes. In some embodiments, the antisense oligonucleotide does not cause neuronal toxicity. In some embodiments, the antisense oligonucleotide does not bind to genomic sequences other than the sequence shown in SEQ ID NO: 1. In some embodiments, the antisense oligonucleotide does not have off-target effects. In some embodiments, the antisense oligonucleotide comprises or consists of the sequence shown in any one of SEQ ID NOs: 20-355, 400-638, 810-959 or 961-1332.

[0252] In some embodiments, the antisense oligonucleotides described herein reduce the expression of UNC13A cryptic exon mRNA. In some cases, this reduction of UNC13A cryptic exon mRNA alone does not indicate an upregulation of full-length UNC13A mRNA. In some cases, the antisense oligonucleotides interfere with the correct splicing of introns.

[0253] In some embodiments, the antisense oligonucleotides occupy overlapping regions with other antisense oligonucleotides described herein. In some cases, two antisense oligonucleotides targeting adjacent and / or overlapping regions of UNC13A mRNA have different activities, wherein the first antisense oligonucleotide is active and the second antisense oligonucleotide is inactive. In some cases, two antisense oligonucleotides targeting adjacent and / or overlapping regions of UNC13A mRNA have the same nucleotide length or different nucleotide lengths. In some cases, a single nucleotide shift in the target sequence affects the ability of the antisense oligonucleotide to block the splicing mechanism. In some cases, a single nucleotide shift in the target sequence results in significantly different antisense oligonucleotide activities, wherein the first antisense oligonucleotide is active and the second antisense oligonucleotide is inert.

[0254] In some cases, the antisense oligonucleotides described herein that consist of at least 18 nucleotides are more active. In some cases, antisense oligonucleotides consisting of at least 20 nucleotides are the most effective, particularly those consisting of 20, 21, or 22 nucleotides. In some cases, selected antisense oligonucleotides consisting of 13 to 17 nucleotides, particularly 13 or 15 nucleotides, are active. In some embodiments, this indicates the minimum region that needs to be blocked to have the desired effect. This highlights the importance of both the location and length of the antisense oligonucleotide on UNC13A mRNA for optimal performance.

[0255] In some cases, the antisense oligonucleotides are provided to the cells for a continuously extended period of time. In some cases, the antisense oligonucleotides restore UNC13A levels to those of wild-type cells. In some cases, the cells are neurons from one or more different iPSC cell lines, such as GM25256 (GM or GM cell line) and NDS00262 (005 or 005 cell line).

[0256] III. Pharmaceutical Compositions

[0257] In some aspects, the present disclosure provides a pharmaceutical composition comprising a UNC13A cryptic exon splicing variant specific inhibitor as described herein and a pharmaceutically acceptable carrier. As used herein, the term "pharmaceutically acceptable" refers to those compounds, materials, compositions, and / or dosage forms that are suitable, within the scope of sound medical judgment, for use in contact with cells and / or tissues without excessive toxicity, irritation, allergic response, or other problems or complications and are commensurate with a reasonable benefit / risk ratio.

[0258] As used herein, the term "pharmaceutically acceptable carrier" means a pharmaceutically acceptable material, composition, or vehicle, such as a liquid or solid filler, stabilizer, dispersing agent, suspending agent, diluent, excipient, thickening agent, solvent, or encapsulating material, involved in carrying or transporting a compound useful in the present invention within a patient or to a patient such that the compound can perform its intended function. Each carrier must be "acceptable" in the sense of being compatible with the other ingredients of the formulation and not injurious to the contacted cells or tissues. Additional components that may be included in the pharmaceutical compositions used in the practice of the present invention are known in the art and are described in, for example, Remington's Pharmaceutical Sciences (Genaro, editor, Mack Publishing Co., Easton, PA, 1985), which is incorporated herein by reference.

[0259] As is well known in the medical arts, the dosage for any given patient depends on a number of factors, including the patient's size, weight, body surface area, age, the level of the UNC13A cryptic exon splicing variant specific inhibitor required to achieve a therapeutic effect, the stability of the UNC13A cryptic exon splicing variant specific inhibitor, the particular disease being treated, the stage of the disease, sex, time and route of administration, general health, and other drugs being administered concurrently.

[0260] As determined by one of ordinary skill in the art, the pharmaceutical composition can be administered in a manner suitable for the disease or condition to be treated (or prevented). The appropriate dosage of the composition and the appropriate duration and frequency of administration will be determined by such factors as the patient's health status, the patient's body size (i.e., weight, mass, or body surface area), the type and severity of the patient's disease, the specific form of the active ingredient, and the method of administration. Generally, the appropriate dosage and treatment regimen provide the composition in an amount sufficient to provide a therapeutic and / or prophylactic benefit (as described herein, including improved clinical outcomes such as more frequent complete or partial remission, or longer disease-free and / or overall survival or reduced symptom severity). For prophylactic use, the dosage should be sufficient to prevent, delay the onset of, or reduce the severity of a disease associated with a disorder or condition. The prophylactic benefit of the composition administered according to the methods described herein can be determined by conducting preclinical (including in vitro and in vivo animal studies) and clinical studies and analyzing the data obtained therefrom by appropriate statistical, biological, and clinical methods and techniques, all of which can be readily practiced by one of ordinary skill in the art.

[0261] The composition (e.g., pharmaceutical composition) can be administered by any route, including enteral (e.g., oral), parenteral, intravenous, intramuscular, intraarterial, intramedullary, intrathecal, subpial, parenchymal, intracerebral, intraventricular, intraocular, intraventricular, lumbar, subcutaneous, transdermal, intradermal, rectal, intravaginal, intraperitoneal, topical (such as by powder, ointment, cream, and / or drops), mucosal, nasal, buccal, sublingual; by intratracheal instillation, bronchial instillation, and / or inhalation; and / or as an oral spray, nasal spray, and / or aerosol. Generally, the most appropriate route of administration will depend on various factors, including the nature of the agent (e.g., its stability in the gastrointestinal environment) and / or the condition of the subject. In some embodiments, the composition is directly injected into the CNS of the subject. In some embodiments, the direct injection into the CNS is intracerebral injection, parenchymal injection, intrathecal injection, intracerebral injection, subpial injection, or any combination thereof. In some embodiments, the direct injection into the CNS is directly injected into the cerebrospinal fluid (CSF) of the subject, optionally wherein the direct injection is intracisternal injection, intraventricular injection, and / or lumbar injection.

[0262] IV. Methods of Using UNC13A Cryptic Splice Variant Inhibitors

[0263] The antisense oligonucleotides provided herein have many beneficial properties that make them useful for increasing UNC13A protein expression, reducing the inclusion of UNC13A cryptic exons, and treating neurodegenerative diseases, as further described below.

[0264] A. Methods of Using ASOs to Increase UNC13A Protein Expression

[0265] The present invention provides a method for increasing the expression of UNC13A protein in cells, the method comprising contacting the cells with an antisense oligonucleotide provided herein. In some embodiments, after the cells are contacted with the antisense oligonucleotide, the expression of UNC13A protein is increased by at least 10%. In some embodiments, the expression of UNC13A protein is increased by at least 20%, at least 30%, at least 40%, at least 50% or at least 75% compared to the level before contact with the antisense oligonucleotide. In some embodiments, the expression of UNC13A is increased by at least 1.1-fold, at least 1.2-fold, at least 1.3-fold, at least 1.5-fold, at least 1.5-fold or at least 2-fold compared to the level before contact with the antisense oligonucleotide. In some embodiments, the amount of full-length UNC13A protein is increased. In some embodiments, the ASO targets regions 1-4 of the UNC13A mRNA transcript identified herein. In some embodiments, the ASO targets region 2 or 3 of the UNC13A mRNA transcript. In some embodiments, the antisense oligonucleotide comprises or consists of the sequences shown in SEQ ID NO: 20-355 or SEQ ID NO: 20-40 or SEQ ID NO: 41-355 or SEQ ID NO: 810-957. In some embodiments, the antisense oligonucleotide comprises or consists of any one of the sequences shown in SEQ ID NO: 20-355, 400-638, 810-959 or 961-1332. In some embodiments, the antisense oligonucleotide comprises or consists of any one of the sequences shown in SEQ ID NO: 1059-1074, 1075-1089, 1090-1103, 1248-1270, 1271-1292 and 1293-1313. In some embodiments, the antisense oligonucleotide comprises or consists of any one of the sequences shown in SEQ ID NO: 1059-1074, 1075-1089 and 1090-1103. In some embodiments, the antisense oligonucleotide comprises or consists of any one of the sequences shown in SEQ ID NO: 1248-1270, 1271-1292 and 1293-1313. In some embodiments, the antisense oligonucleotide has a modification.

[0266] In some embodiments, after the cells are contacted with the antisense oligonucleotide, the expression of the UNC13A protein increases to a normal level. In some embodiments, the expression of the UNC13A protein increases to the level of an individual without a mutation in TDP-43. In some embodiments, the expression of the UNC13A protein increases to the level of an individual without a decrease in TDP-43 protein expression. In some embodiments, the expression of the UNC13A protein increases to the level of an individual without a risk allele associated with ALS or FTD. In some embodiments, the expression of UNC13A increases to the level of an individual without ALS or FTD. In some embodiments, the antisense oligonucleotide comprises or consists of the sequences shown in SEQ ID NO: 20-355 or SEQ ID NO: 20-40 or SEQ ID NO: 41-355 or SEQ ID NO: 810-957. In some embodiments, the antisense oligonucleotide comprises or consists of the sequences shown in any one of SEQ ID NO: 20-355, 400-638, 810-959, or 961-1332. In some embodiments, the antisense oligonucleotide comprises or consists of the sequences shown in any one of SEQ ID NO: 1059-1074, 1075-1089, 1090-1103, 1248-1270, 1271-1292, and 1293-1313. In some embodiments, the antisense oligonucleotide comprises or consists of the sequences shown in any one of SEQ ID NO: 1059-1074, 1075-1089, and 1090-1103. In some embodiments, the antisense oligonucleotide comprises or consists of the sequences shown in any one of SEQ ID NO: 1248-1270, 1271-1292, and 1293-1313. In some embodiments, the antisense oligonucleotide has a modification.

[0267] In some embodiments, prior to contact of the cell with the antisense oligonucleotide, the cell has a low level of UNC13A protein expression. In some embodiments, the cell has a mutation in TDP-43 or reduced expression of the TDP-43 protein. In some embodiments, the cell has a level of UNC13A protein expression associated with a TDP-43 proteinopathy. In some embodiments, the cell has a level of UNC13A protein associated with a risk allele associated with ALS or FTD. In some embodiments, the cell has a level of UNC13A associated with ALS or FTD. In some embodiments, the antisense oligonucleotide comprises or consists of the sequences shown in SEQ ID NO: 20-355 or SEQ ID NO: 20-40 or SEQ ID NO: 41-355 or SEQ ID NO: 810-957. In some embodiments, the antisense oligonucleotide comprises or consists of the sequences shown in any one of SEQ ID NO: 20-355, 400-638, 810-959 or 961-1332. In some embodiments, the antisense oligonucleotide has a modification.

[0268] In some embodiments, compared to normal or wild-type cells, prior to contact of the cell with the antisense oligonucleotide, the cell has a reduced level of UNC13A protein expression. In some embodiments, normal cells are cells that do not have a mutation in TDP-43, have a normal expression level of the TDP-43 protein level, do not have a risk allele associated with ALS or FTD, and / or are not obtained from an individual with ALSO or FTD. In some embodiments, prior to contact with the ASO, the cell has an undetectable level of UNC13A protein expression. In some embodiments, the cell expresses less than 80%, less than 70%, less than 60% or less than 50% of the normal level of the UNC13A protein. In some embodiments, the antisense oligonucleotide comprises or consists of the sequences shown in SEQ ID NO: 20-355 or SEQ ID NO: 20-40 or SEQ ID NO: 41-355 or SEQ ID NO: 810-957. In some embodiments, the antisense oligonucleotide comprises or consists of the sequences shown in any one of SEQ ID NO: 20-355, 400-638, 810-959 or 961-1332. In some embodiments, the antisense oligonucleotide has a modification.

[0269] In some aspects, provided herein is a method of increasing the expression of UNC13A protein in a cell by at least 10%, the method comprising contacting the cell with an antisense oligonucleotide comprising 13 to 30 nucleotides, wherein the antisense oligonucleotide is complementary to the sequence shown in SEQ ID NO:1. In some aspects, provided herein is a method of increasing the expression of UNC13A protein in a cell by at least 10%, the method comprising contacting the cell with an antisense oligonucleotide comprising 13 to 30 nucleotides, wherein the antisense oligonucleotide is complementary to the sequence shown in any one of SEQ ID NOs: 1339-1342 or a portion thereof. In some embodiments, the antisense oligonucleotide that modulates UNC13A has a sequence complementary to the sequence of UNC13A mRNA located between the 5' splice donor site and the 3' splice acceptor site of a cryptic exon located between exons 20 and 21 of the UNC13A gene.

[0270] In some embodiments, the antisense oligonucleotide is complementary to the sequence shown in SEQ ID NO:1. In some embodiments, the antisense oligonucleotide has at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 95% or at least 98% complementarity with the sequence shown in SEQ ID NO:1.

[0271] In some embodiments, the antisense oligonucleotide is complementary to the sequence shown in any one of SEQ ID NOs: 700 - 703. In some embodiments, the antisense oligonucleotide is complementary to the sequence shown in SEQ ID NO: 701 or 703. In some embodiments, the antisense oligonucleotide has at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 95% or at least 98% complementarity with the sequence shown in SEQ ID NO: 701 or 703. In some embodiments, the antisense oligonucleotide is completely complementary to the sequence shown in SEQ ID NO: 701 or 703. In some embodiments, the antisense oligonucleotide is complementary to the sequence shown in any one of SEQ ID NOs: 700. In some embodiments, the antisense oligonucleotide has at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 95% or at least 98% complementarity with the sequence shown in SEQ ID NO: 700. In some embodiments, the antisense oligonucleotide is completely complementary to the sequence shown in SEQ ID NO: 700. In some embodiments, the antisense oligonucleotide is complementary to the sequence shown in any one of SEQ ID NOs: 701. In some embodiments, the antisense oligonucleotide has at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 95% or at least 98% complementarity with the sequence shown in SEQ ID NO: 701. In some embodiments, the antisense oligonucleotide is completely complementary to the sequence shown in SEQ ID NO: 701. In some embodiments, the antisense oligonucleotide is complementary to the sequence shown in any one of SEQ ID NOs: 702. In some embodiments, the antisense oligonucleotide has at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 95% or at least 98% complementarity with the sequence shown in SEQ ID NO: 702. In some embodiments, the antisense oligonucleotide is completely complementary to the sequence shown in SEQ ID NO: 702. In some embodiments, the antisense oligonucleotide is complementary to the sequence shown in any one of SEQ ID NOs: 703. In some embodiments, the antisense oligonucleotide has at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 95% or at least 98% complementarity with the sequence shown in SEQ ID NO: 703. In some embodiments, the antisense oligonucleotide is completely complementary to the sequence shown in SEQ ID NO: 703.

[0272] In some embodiments, the antisense oligonucleotide comprises at least 80%, at least 90%, at least 95%, at least 98% or 100% complementarity to the sequences shown in SEQ ID NO: 700-703. In some embodiments, the antisense oligonucleotide comprises at least 80%, at least 90%, at least 95%, at least 98% or 100% complementarity to the sequence shown in SEQ ID NO: 701. In some embodiments, the antisense oligonucleotide comprises at least 80%, at least 90%, at least 95%, at least 98% or 100% complementarity to the sequence shown in SEQ ID NO: 703.

[0273] In some cases, the methods provided herein comprise contacting a cell with an antisense oligonucleotide that is complementary to a sequence of UNC13A mRNA, wherein the sequence of the UNC13A mRNA is between the 5' splice donor site and the 3' splice acceptor site of a cryptic exon located between exons 20 and 21 of the UNC13A gene, and wherein the sequence of UNC13A is uniquely sensitive to antisense oligonucleotide binding. In some cases, regions of the UNC13A mRNA are identified as being more responsive than other regions. Here, these regions are determined to be sensitive to a variety of factors, including but not limited to antisense oligonucleotide length, composition, and localization on the UNC13A mRNA. The smallest regions of the UNC13A mRNA are specifically targeted to maximize the activity of antisense oligonucleotides that regulate UNC13A mRNA splicing. In some embodiments, these highly responsive smallest regions of the UNC13A mRNA are referred to as hotspots.

[0274] In some embodiments, the minimal region of UNC13A mRNA between the 5' splice donor site and the 3' splice acceptor site of a cryptic exon located between exons 20 and 21 of the UNC13A gene is identified as being highly responsive to binding with an antisense oligonucleotide or another similar nucleic acid probe. In some embodiments, an ASO that binds to such a minimal region is effective in preventing the inclusion of the UNC13A cryptic exon. In some embodiments, the minimal region is 3 to 30, 4 to 26, 5 to 22, 6 to 18, 7 to 14, or 8 to 10 nucleotides in length. In some embodiments, the minimal region is no more than 30, no more than 25, no more than 20, or no more than 15 nucleotides in length. In some embodiments, the minimal region is 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 nucleotides in length. In some embodiments, the minimal region is 18 nucleotides in length. In some embodiments, the minimal region is 29 nucleotides in length. In some embodiments, the minimal region comprises the sequence shown in any of SEQ ID NOs: 1339-1342 or a portion thereof. In some embodiments, the minimal region comprises the sequence shown in SEQ ID NO: 1339 or a portion thereof. In some embodiments, the minimal region comprises the sequence shown in SEQ ID NO: 1340 or a portion thereof. In some embodiments, the minimal region comprises the sequence shown in SEQ ID NO: 1341 or a portion thereof. In some embodiments, the minimal region comprises the sequence shown in SEQ ID NO: 1342 or a portion thereof. In some embodiments, binding of the region or a portion thereof to an antisense oligonucleotide enhances splicing regulatory activity. In some embodiments, binding of the minimal region or a portion thereof to an antisense oligonucleotide increases UNC13A mRNA expression. In some embodiments, binding of the minimal region or a portion thereof to an antisense oligonucleotide decreases UNC13A cryptic exon mRNA expression. In some embodiments, binding of the minimal region or a portion thereof to an antisense oligonucleotide increases UNC13A protein expression.

[0275] In some cases, the methods provided herein involve contacting a cell with an antisense oligonucleotide comprising 13 to 30 nucleotides, wherein the antisense oligonucleotide is complementary to the sequence shown in any one of SEQ ID NOs: 1339 - 1342 or a portion thereof. In some embodiments, the antisense oligonucleotide is complementary to the sequence shown in SEQ ID NO: 1339 or a portion thereof. In some embodiments, the antisense oligonucleotide is complementary to the sequence shown in SEQ ID NO: 1340 or a portion thereof. In some embodiments, the antisense oligonucleotide is complementary to the sequence shown in SEQ ID NO: 1341 or a portion thereof. In some embodiments, the antisense oligonucleotide is complementary to the sequence shown in SEQ ID NO: 1342 or a portion thereof. In some embodiments, the antisense oligonucleotide is complementary to at least 1, at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, at least 11, at least 12, or at least 13 nucleotides of the sequence shown in any one of SEQ ID NOs: 1339 - 1342. In some embodiments, the antisense oligonucleotide is complementary to at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, at least 11, at least 12, or at least 13 consecutive nucleotides of the sequence shown in any one of SEQ ID NOs: 1339 - 1342. In some embodiments, the antisense oligonucleotide is complementary to 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, or 13 nucleotides of the sequence shown in any one of SEQ ID NOs: 1339 - 1342. In some embodiments, the antisense oligonucleotide is complementary to 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, or 13 consecutive nucleotides of the sequence shown in any one of SEQ ID NOs: 1339 - 1342. In some embodiments, the antisense oligonucleotide is at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99%, or 100% complementary to the sequence shown in SEQ ID NO: 1339. In some embodiments, the antisense oligonucleotide is at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99%, or 100% complementary to the sequence shown in SEQ ID NO: 1340. In some embodiments, the antisense oligonucleotide is at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99%, or 100% complementary to the sequence shown in SEQ ID NO: 1341.In some embodiments, the antisense oligonucleotide is at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% complementary to the sequence shown in SEQ ID NO: 1342.

[0276] In some embodiments, the methods provided herein comprise contacting a cell with an antisense oligonucleotide comprising 13 to 30 nucleotides, wherein binding of the antisense oligonucleotide to the sequence or a portion thereof shown in any one of SEQ ID NOs: 1339 - 1342 enhances UNC13A splicing regulatory activity. In some embodiments, binding of the antisense oligonucleotide to the sequence or a portion thereof shown in any one of SEQ ID NOs: 1339 - 1342 increases UNC13A mRNA expression. In some embodiments, the expression of UNC13A mRNA in the cell is increased to at least 25%, at least 30%, at least 35%, at least 40%, at least 45% or at least 50% of the level of UNC13A mRNA expressed in wild - type cells. In some embodiments, the expression of UNC13A mRNA in the cell is increased by about 10% to about 95%, 20% to about 85%, 30% to about 75%, 40% to about 65% or 50% to about 55% of the level of UNC13A mRNA expressed in wild - type cells. In some embodiments, the expression of UNC13A mRNA in the cell is increased by at least 50%, at least 100%, at least 200%, at least 300%, at least 400% or at least 500%. In some embodiments, the expression of UNC13A mRNA in the cell is increased by about 25% to about 1000%, about 50% to about 800%, about 100% to about 700%, about 200% to about 600% or about 300% to about 500%. In some embodiments, binding of the antisense oligonucleotide to the sequence or a portion thereof shown in any one of SEQ ID NOs: 1339 - 1342 decreases the expression of UNC13A cryptic exon mRNA. In some embodiments, the expression of UNC13A cryptic exon mRNA in the cell is reduced by at least 10%, at least 20%, at least 30%, at least 40% or at least 50%. In some embodiments, the expression of UNC13A cryptic exon mRNA in the cell is reduced by about 10% to about 90%, about 20% to about 80%, about 30% to about 70% or about 40% to about 60%.

[0277] In some embodiments, the wild - type cell is a cell from an individual not suffering from amyotrophic lateral sclerosis (ALS) or frontotemporal dementia (FTD). In some embodiments, the wild - type cell is a cell that expresses normal levels of the TDP - 43 protein. In some embodiments, the wild - type cell is a cell that does not produce a transcript of UNC13A containing a cryptic exon.

[0278] In some aspects, the present disclosure provides methods for increasing the expression of UNC13A protein in a cell by at least 10%, the method comprising contacting the cell with an antisense oligonucleotide, wherein the antisense oligonucleotide comprises a sequence shown in any one of SEQ ID NO: 20-355, 400-638, 810-959, or 961-1332. In some embodiments, the antisense oligonucleotide comprises or consists of 13 to 30 nucleotides. In some embodiments, the antisense oligonucleotide comprises or consists of 13 to 30, 15 to 28, 17 to 26, 19 to 24, or 21 to 23 nucleotides. In some embodiments, the antisense oligonucleotide comprises or consists of 13 to 25 nucleotides. In some embodiments, the antisense oligonucleotide comprises or consists of 13, 15, 17, 18, 19, 20, 21, 22, or 24 nucleotides. In some embodiments, the antisense oligonucleotide comprises or consists of 19, 20, 21, or 22 nucleotides. In some embodiments, the antisense oligonucleotide comprises or consists of at least 13, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, or 25 nucleotides. In some embodiments, the antisense oligonucleotide comprises or consists of no more than 25, 24, 23, 22, 21, 20, 19, 18, 17, 16, 15, or 13 nucleotides. In some embodiments, the antisense oligonucleotide comprises or consists of 13 nucleotides. In some embodiments, the antisense oligonucleotide comprises or consists of 15 nucleotides. In some embodiments, the antisense oligonucleotide comprises or consists of 17 nucleotides. In some embodiments, the antisense oligonucleotide comprises or consists of 18 nucleotides. In some embodiments, the antisense oligonucleotide comprises or consists of 19 nucleotides. In some embodiments, the antisense oligonucleotide comprises or consists of 20 nucleotides. In some embodiments, the antisense oligonucleotide comprises or consists of 21 nucleotides. In some embodiments, the antisense oligonucleotide comprises or consists of 22 nucleotides. In some embodiments, the antisense oligonucleotide comprises or consists of 23 nucleotides. In some embodiments, the antisense oligonucleotide comprises or consists of 24 nucleotides. In some embodiments, the antisense oligonucleotide comprises or consists of 25 nucleotides. In some embodiments, wherein the antisense oligonucleotide is provided to the cell, the expression of UNC13A mRNA in the cell is increased to at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, or at least 50% of the level of UNC13A mRNA expressed in wild-type cells. In some embodiments, the increase in the expression of UNC13A mRNA in the cell is about 10% to about 95%, 20% to about 85%, 30% to about 75%, 40% to about 65%, or 50% to about 55% of the level of UNC13A mRNA expressed in wild-type cells.In some embodiments, the expression of UNC13A mRNA in the cell is increased by about 25% to about 1000%, about 50% to about 800%, about 100% to about 700%, about 200% to about 600%, or about 300% to about 500%. In some embodiments, wherein an antisense oligonucleotide is provided to the cell, the expression of UNC13A cryptic exon mRNA in the cell is decreased by about 10% to about 90%, about 20% to about 80%, about 30% to about 70%, or about 40% to about 60%. In some embodiments, wherein an antisense oligonucleotide is provided to the cell, the expression of UNC13A protein in the cell is increased to at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, or at least 50% of the level of UNC13A protein expressed in wild-type cells. In some embodiments, the expression of UNC13A protein in the cell is increased by about 10% to about 95%, 20% to about 85%, 30% to about 75%, 40% to about 65%, or 50% to about 55% of the level of UNC13A protein expressed in wild-type cells. In some embodiments, the expression of UNC13A protein in the cell is increased by about 25% to about 1000%, about 50% to about 800%, about 100% to about 700%, about 200% to about 600%, or about 300% to about 500%. In some embodiments, wherein an antisense oligonucleotide is provided to the cell, the expression of UNC13A protein is increased to a level sufficient to improve the function of one or more neurons selected from the group consisting of: neuronal signaling, neuronal morphology, neurite length, neurotransmitter release, action potential, neuronal synaptic plasticity, and neuronal lifespan. In some embodiments, the antisense oligonucleotide consists of the sequence shown in any one of SEQ ID NO: 1059-1074, 1075-1089, 1090-1103, 1248-1270, 1271-1292, and 1293-1313. In some embodiments, the antisense oligonucleotide consists of the sequence shown in any one of SEQ ID NO: 1059-1074, 1075-1089, and 1090-1103. In some embodiments, the antisense oligonucleotide consists of the sequence shown in any one of SEQ ID NO: 1248-1270, 1271-1292, and 1293-1313. In some embodiments, the antisense oligonucleotide comprises a modification. In some embodiments, the modification is selected from the group consisting of: 2'-OMe antisense oligonucleotide, 2'-O-methoxyethyl antisense oligonucleotide, phosphorothioate antisense oligonucleotide, and LNA antisense oligonucleotide.

[0279] In some embodiments, the method of increasing the expression of UNC13A protein in a cell by at least 10% comprises contacting the cell with an antisense oligonucleotide, wherein the antisense oligonucleotide is complementary to the sequence shown in SEQ ID NO:1. In some embodiments, the antisense oligonucleotide comprises or consists of the sequence shown in any one of SEQ ID NO:20-355, 400-638, 810-959 or 961-1332. In some embodiments, the antisense oligonucleotide comprises or consists of 13 to 30 nucleotides. In some embodiments, the antisense oligonucleotide comprises a modification.

[0280] In some embodiments, the method of increasing the expression of UNC13A protein in a cell by at least 10% comprises contacting the cell with an antisense oligonucleotide, wherein the antisense oligonucleotide consists of the sequence shown in any one of SEQ ID NO:20-355, 400-638, 810-959 or 961-1332. In some embodiments, the antisense oligonucleotide comprises or consists of 13 to 30 nucleotides. In some embodiments, the antisense oligonucleotide comprises a modification.

[0281] In some embodiments, the method of increasing the expression of UNC13A protein in a cell by at least 10% comprises contacting the cell with an antisense oligonucleotide, wherein the antisense oligonucleotide comprises 13 nucleotides complementary to the sequence shown in SEQ ID NO:1, and wherein the antisense oligonucleotide comprises the sequence shown in any of SEQ ID NOs: 961 - 983 and 1116 - 1145. In some embodiments, the antisense oligonucleotide consists of 13 nucleotides complementary to the sequence shown in SEQ ID NO:1, and wherein the antisense oligonucleotide consists of the sequence shown in any of SEQ ID NOs: 961 - 983 and 1116 - 1145. In some embodiments, wherein the antisense oligonucleotide is provided to the cell, the expression of UNC13A mRNA in the cell is increased to at least 25%, at least 30%, at least 35%, at least 40%, at least 45% or at least 50% of the level of UNC13A mRNA expressed in wild-type cells. In some embodiments, the increase in the expression of UNC13A mRNA in the cell is from about 10% to about 95%, 20% to about 85%, 30% to about 75%, 40% to about 65% or 50% to about 55% of the level of UNC13A mRNA expressed in wild-type cells. In some embodiments, the increase in the expression of UNC13A mRNA in the cell is from about 25% to about 1000%, about 50% to about 800%, about 100% to about 700%, about 200% to about 600% or about 300% to about 500%. In some embodiments, wherein the antisense oligonucleotide is provided to the cell, the expression of UNC13A cryptic exon mRNA in the cell is decreased by about 10% to about 90%, about 20% to about 80%, about 30% to about 70% or about 40% to about 60%. In some embodiments, wherein the antisense oligonucleotide is provided to the cell, the expression of UNC13A protein in the cell is increased to at least 25%, at least 30%, at least 35%, at least 40%, at least 45% or at least 50% of the level of UNC13A protein expressed in wild-type cells. In some embodiments, the increase in the expression of UNC13A protein in the cell is from about 10% to about 95%, 20% to about 85%, 30% to about 75%, 40% to about 65% or 50% to about 55% of the level of UNC13A protein expressed in wild-type cells. In some embodiments, the increase in the expression of UNC13A protein in the cell is from about 25% to about 1000%, about 50% to about 800%, about 100% to about 700%, about 200% to about 600% or about 300% to about 500%. In some embodiments, wherein the antisense oligonucleotide is provided to the cell, the expression of UNC13A protein is increased to a level sufficient to improve the function of one or more neurons selected from the group consisting of: neuronal signaling, neuronal morphology, neurite length, neurotransmitter release, action potential, neuronal synaptic plasticity and neuronal lifespan.In some embodiments, the antisense oligonucleotide consists of the sequence shown in any one of SEQ ID NO: 1059 - 1074, 1075 - 1089, 1090 - 1103, 1248 - 1270, 1271 - 1292, and 1293 - 1313. In some embodiments, the antisense oligonucleotide consists of the sequence shown in any one of SEQ ID NO: 1059 - 1074, 1075 - 1089, and 1090 - 1103. In some embodiments, the antisense oligonucleotide consists of the sequence shown in any one of SEQ ID NO: 1248 - 1270, 1271 - 1292, and 1293 - 1313. In some embodiments, the antisense oligonucleotide comprises a modification. In some embodiments, the modification is selected from the group consisting of: 2'-OMe antisense oligonucleotide, 2'-O-methoxyethyl antisense oligonucleotide, phosphorothioate antisense oligonucleotide, and LNA antisense oligonucleotide.

[0282] In some embodiments, the method of increasing the expression of UNC13A protein in a cell by at least 10% comprises contacting the cell with an antisense oligonucleotide, wherein the antisense oligonucleotide comprises 15 nucleotides complementary to the sequence shown in SEQ ID NO:1, and wherein the antisense oligonucleotide comprises the sequence shown in any of SEQ ID NOs: 984 - 1004 and 1146 - 1173. In some embodiments, the antisense oligonucleotide consists of 15 nucleotides complementary to the sequence shown in SEQ ID NO:1, and wherein the antisense oligonucleotide consists of the sequence shown in any of SEQ ID NOs: 984 - 1004 and 1146 - 1173. In some embodiments, wherein an antisense oligonucleotide is provided to a cell, the expression of UNC13A mRNA in the cell is increased to at least 25%, at least 30%, at least 35%, at least 40%, at least 45% or at least 50% of the level of UNC13A mRNA expressed in wild - type cells. In some embodiments, the increase in the expression of UNC13A mRNA in the cell is from about 10% to about 95%, 20% to about 85%, 30% to about 75%, 40% to about 65% or 50% to about 55% of the level of UNC13A mRNA expressed in wild - type cells. In some embodiments, the increase in the expression of UNC13A mRNA in the cell is from about 25% to about 1000%, about 50% to about 800%, about 100% to about 700%, about 200% to about 600% or about 300% to about 500%. In some embodiments, wherein an antisense oligonucleotide is provided to a cell, the expression of UNC13A cryptic exon mRNA in the cell is reduced by about 10% to about 90%, about 20% to about 80%, about 30% to about 70% or about 40% to about 60%. In some embodiments, wherein an antisense oligonucleotide is provided to a cell, the expression of UNC13A protein in the cell is increased to at least 25%, at least 30%, at least 35%, at least 40%, at least 45% or at least 50% of the level of UNC13A protein expressed in wild - type cells. In some embodiments, the increase in the expression of UNC13A protein in the cell is from about 10% to about 95%, 20% to about 85%, 30% to about 75%, 40% to about 65% or 50% to about 55% of the level of UNC13A protein expressed in wild - type cells. In some embodiments, the increase in the expression of UNC13A protein in the cell is from about 25% to about 1000%, about 50% to about 800%, about 100% to about 700%, about 200% to about 600% or about 300% to about 500%. In some embodiments, wherein an antisense oligonucleotide is provided to a cell, the expression of UNC13A protein is increased to a level sufficient to improve the function of one or more neurons selected from the group consisting of: neuronal signaling, neuronal morphology, neurite length, neurotransmitter release, action potential, neuronal synaptic plasticity, and neuronal lifespan.In some embodiments, the antisense oligonucleotide consists of the sequence shown in any one of SEQ ID NO: 1059 - 1074, 1075 - 1089, 1090 - 1103, 1248 - 1270, 1271 - 1292, and 1293 - 1313. In some embodiments, the antisense oligonucleotide consists of the sequence shown in any one of SEQ ID NO: 1059 - 1074, 1075 - 1089, and 1090 - 1103. In some embodiments, the antisense oligonucleotide consists of the sequence shown in any one of SEQ ID NO: 1248 - 1270, 1271 - 1292, and 1293 - 1313. In some embodiments, the antisense oligonucleotide comprises a modification. In some embodiments, the modification is selected from the group consisting of: 2'-OMe antisense oligonucleotide, 2'-O-methoxyethyl antisense oligonucleotide, phosphorothioate antisense oligonucleotide, and LNA antisense oligonucleotide.

[0283] In some embodiments, the method of increasing the expression of UNC13A protein in a cell by at least 10% comprises contacting the cell with an antisense oligonucleotide, wherein the antisense oligonucleotide comprises 16 nucleotides complementary to the sequence shown in SEQ ID NO:1, and wherein the antisense oligonucleotide comprises the sequence shown in any one of SEQ ID NO:41, 42, 56, 57, 71, 72, 86, 87, 101, 102, 116, 117, 131, 132, 146, 147, 161, 162, 176, 177, 191, 192, 206, 207, 221, 222, 236, 237, 251, 252, 266, 267, 281, 282, 296, 297, 311, 326, 341, 810, 811, 812, 813, 826, 827, 835, 838, 841, 842, 851, 852, 861, 864, 867, 868, 877, 878, 895, 900, 901, 910, 934 and 935. In some embodiments, the antisense oligonucleotide consists of 16 nucleotides complementary to the sequence shown in SEQ ID NO:1, and wherein the antisense oligonucleotide consists of the sequence shown in any one of SEQ ID NO:41, 42, 56, 57, 71, 72, 86, 87, 101, 102, 116, 117, 131, 132, 146, 147, 161, 162, 176, 177, 191, 192, 206, 207, 221, 222, 236, 237, 251, 252, 266, 267, 281, 282, 296, 297, 311, 326, 341, 810, 811, 812, 813, 826, 827, 835, 838, 841, 842, 851, 852, 861, 864, 867, 868, 877, 878, 895, 900, 901, 910, 934 and 935. In some embodiments, wherein the antisense oligonucleotide is provided to the cell, the expression of UNC13A mRNA in the cell is increased to at least 25%, at least 30%, at least 35%, at least 40%, at least 45% or at least 50% of the level of UNC13A mRNA expressed in wild-type cells. In some embodiments, the increase in the expression of UNC13A mRNA in the cell is from about 10% to about 95%, 20% to about 85%, 30% to about 75%, 40% to about 65% or 50% to about 55% of the level of UNC13A mRNA expressed in wild-type cells. In some embodiments, the increase in the expression of UNC13A mRNA in the cell is about 25% to about 1000%, about 50% to about 800%, about 100% to about 700%, about 200% to about 600% or about 300% to about 500%.In some embodiments, where an antisense oligonucleotide is provided to a cell, the expression of UNC13A cryptic exon mRNA in the cell is reduced by about 10% to about 90%, about 20% to about 80%, about 30% to about 70%, or about 40% to about 60%. In some embodiments, where an antisense oligonucleotide is provided to a cell, the expression of UNC13A protein in the cell is increased to at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, or at least 50% of the level of UNC13A protein expressed in wild-type cells. In some embodiments, the expression of UNC13A protein in the cell is increased by about 10% to about 95%, 20% to about 85%, 30% to about 75%, 40% to about 65%, or 50% to about 55% of the level of UNC13A protein expressed in wild-type cells. In some embodiments, the expression of UNC13A protein in the cell is increased by about 25% to about 1000%, about 50% to about 800%, about 100% to about 700%, about 200% to about 600%, or about 300% to about 500%. In some embodiments, where an antisense oligonucleotide is provided to a cell, the expression of UNC13A protein is increased to a level sufficient to improve the function of one or more neurons selected from the group consisting of: neuronal signaling, neuronal morphology, neurite length, neurotransmitter release, action potential, neuronal synaptic plasticity, and neuronal lifespan. In some embodiments, the antisense oligonucleotide consists of the sequence shown in any one of SEQ ID NO: 1059-1074, 1075-1089, 1090-1103, 1248-1270, 1271-1292, and 1293-1313. In some embodiments, the antisense oligonucleotide consists of the sequence shown in any one of SEQ ID NO: 1059-1074, 1075-1089, and 1090-1103. In some embodiments, the antisense oligonucleotide consists of the sequence shown in any one of SEQ ID NO: 1248-1270, 1271-1292, and 1293-1313. In some embodiments, the antisense oligonucleotide comprises a modification. In some embodiments, the modification is selected from the group consisting of: 2' OMe antisense oligonucleotide, 2'-O-methoxyethyl antisense oligonucleotide, phosphorothioate antisense oligonucleotide, and LNA antisense oligonucleotide.

[0284] In some embodiments, the method of increasing the expression of UNC13A protein in a cell by at least 10% comprises contacting the cell with an antisense oligonucleotide, wherein the antisense oligonucleotide comprises 17 nucleotides complementary to the sequence shown in SEQ ID NO:1, and wherein the antisense oligonucleotide comprises the sequence shown in any one of SEQ ID NO:43, 58, 73, 88, 103, 118, 133, 148, 163, 178, 193, 208, 223, 238, 253, 268, 283, 298, 312, 313, 327, 328, 342, 343, 814, 815, 816, 828, 829, 836, 839, 843, 844, 853, 854, 862, 865, 869, 870, 879, 880, 881, 891, 896, 897, 902, 903, 1005 - 1023 and 1174 - 1199. In some embodiments, the antisense oligonucleotide consists of 17 nucleotides complementary to the sequence shown in SEQ ID NO:1, and wherein the antisense oligonucleotide consists of the sequence shown in any one of SEQ ID NO:43, 58, 73, 88, 103, 118, 133, 148, 163, 178, 193, 208, 223, 238, 253, 268, 283, 298, 312, 313, 327, 328, 342, 343, 814, 815, 816, 828, 829, 836, 839, 843, 844, 853, 854, 862, 865, 869, 870, 879, 880, 881, 891, 896, 897, 902, 903, 1005 - 1023 and 1174 - 1199. In some embodiments, wherein the antisense oligonucleotide is provided to the cell, the expression of UNC13A mRNA in the cell is increased to at least 25%, at least 30%, at least 35%, at least 40%, at least 45% or at least 50% of the level of UNC13A mRNA expressed in wild - type cells. In some embodiments, the increase in the expression of UNC13A mRNA in the cell is about 10% to about 95%, 20% to about 85%, 30% to about 75%, 40% to about 65% or 50% to about 55% of the level of UNC13A mRNA expressed in wild - type cells. In some embodiments, the increase in the expression of UNC13A mRNA in the cell is about 25% to about 1000%, about 50% to about 800%, about 100% to about 700%, about 200% to about 600% or about 300% to about 500%. In some embodiments, wherein the antisense oligonucleotide is provided to the cell, the expression of UNC13A cryptic exon mRNA in the cell is reduced by about 10% to about 90%, about 20% to about 80%, about 30% to about 70% or about 40% to about 60%.In some embodiments, where an antisense oligonucleotide is provided to a cell, the expression of UNC13A protein in the cell is increased to at least 25%, at least 30%, at least 35%, at least 40%, at least 45% or at least 50% of the level of UNC13A protein expressed in wild-type cells. In some embodiments, the increase in the expression of UNC13A protein in the cell is from about 10% to about 95%, 20% to about 85%, 30% to about 75%, 40% to about 65% or 50% to about 55% of the level of UNC13A protein expressed in wild-type cells. In some embodiments, the increase in the expression of UNC13A protein in the cell is from about 25% to about 1000%, about 50% to about 800%, about 100% to about 700%, about 200% to about 600% or about 300% to about 500%. In some embodiments, where an antisense oligonucleotide is provided to a cell, the expression of UNC13A protein is increased to a level sufficient to improve the function of one or more neurons selected from the group consisting of: neuronal signaling, neuronal morphology, neurite length, neurotransmitter release, action potential, neuronal synaptic plasticity and neuronal lifespan. In some embodiments, the antisense oligonucleotide consists of a sequence shown in any one of SEQ ID NO: 1059-1074, 1075-1089, 1090-1103, 1248-1270, 1271-1292 and 1293-1313. In some embodiments, the antisense oligonucleotide consists of a sequence shown in any one of SEQ ID NO: 1059-1074, 1075-1089 and 1090-1103. In some embodiments, the antisense oligonucleotide consists of a sequence shown in any one of SEQ ID NO: 1248-1270, 1271-1292 and 1293-1313. In some embodiments, the antisense oligonucleotide comprises a modification. In some embodiments, the modification is selected from the group consisting of: 2'OMe antisense oligonucleotide, 2'O-methoxyethyl antisense oligonucleotide, phosphorothioate antisense oligonucleotide and LNA antisense oligonucleotide.

[0285] In some embodiments, the method of increasing the expression of UNC13A protein in a cell by at least 10% comprises contacting the cell with an antisense oligonucleotide, wherein the antisense oligonucleotide comprises 18 nucleotides complementary to the sequence shown in SEQ ID NO:1, and wherein the antisense oligonucleotide comprises any one of the sequences shown in SEQ ID NO:38 - 40, 44, 45, 59, 60, 74, 75, 89, 90, 104, 105, 119, 120, 134, 135, 149, 150, 164, 165, 179, 180, 194, 195, 209, 210, 224, 225, 239, 240, 254, 255, 269, 270, 284, 285, 299, 300, 314, 329, 344, 518 - 638, 817, 818, 845, 855, 871, 882, 904, 958, 959, 960, 1024 - 1041, and 1200 - 1224. In some embodiments, the antisense oligonucleotide consists of 18 nucleotides complementary to the sequence shown in SEQ ID NO:1, and wherein the antisense oligonucleotide consists of any one of the sequences shown in SEQ ID NO:38 - 40, 44, 45, 59, 60, 74, 75, 89, 90, 104, 105, 119, 120, 134, 135, 149, 150, 164, 165, 179, 180, 194, 195, 209, 210, 224, 225, 239, 240, 254, 255, 269, 270, 284, 285, 299, 300, 314, 329, 344, 518 - 638, 817, 818, 845, 855, 871, 882, 904, 958, 959, 960, 1024 - 1041, and 1200 - 1224. In some embodiments, wherein the antisense oligonucleotide is provided to the cell, the expression of UNC13A mRNA in the cell is increased to at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, or at least 50% of the level of UNC13A mRNA expressed in wild - type cells. In some embodiments, the increase in the expression of UNC13A mRNA in the cell is from about 10% to about 95%, 20% to about 85%, 30% to about 75%, 40% to about 65%, or 50% to about 55% of the level of UNC13A mRNA expressed in wild - type cells. In some embodiments, the increase in the expression of UNC13A mRNA in the cell is about 25% to about 1000%, about 50% to about 800%, about 100% to about 700%, about 200% to about 600%, or about 300% to about 500%.In some embodiments, where an antisense oligonucleotide is provided to a cell, the expression of the UNC13A cryptic exon mRNA in the cell is reduced by about 10% to about 90%, about 20% to about 80%, about 30% to about 70%, or about 40% to about 60%. In some embodiments, where an antisense oligonucleotide is provided to a cell, the expression of the UNC13A protein in the cell is increased to at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, or at least 50% of the level of the UNC13A protein expressed in wild-type cells. In some embodiments, the expression of the UNC13A protein in the cell is increased by about 10% to about 95%, 20% to about 85%, 30% to about 75%, 40% to about 65%, or 50% to about 55% of the level of the UNC13A protein expressed in wild-type cells. In some embodiments, the expression of the UNC13A protein in the cell is increased by about 25% to about 1000%, about 50% to about 800%, about 100% to about 700%, about 200% to about 600%, or about 300% to about 500%. In some embodiments, where an antisense oligonucleotide is provided to a cell, the expression of the UNC13A protein is increased to a level sufficient to improve the function of one or more neurons selected from the group consisting of: neuronal signaling, neuronal morphology, neurite length, neurotransmitter release, action potential, neuronal synaptic plasticity, and neuronal lifespan. In some embodiments, the antisense oligonucleotide consists of the sequence shown in any one of SEQ ID NO: 1059-1074, 1075-1089, 1090-1103, 1248-1270, 1271-1292, and 1293-1313. In some embodiments, the antisense oligonucleotide consists of the sequence shown in any one of SEQ ID NO: 1059-1074, 1075-1089, and 1090-1103. In some embodiments, the antisense oligonucleotide consists of the sequence shown in any one of SEQ ID NO: 1248-1270, 1271-1292, and 1293-1313. In some embodiments, the antisense oligonucleotide comprises a modification. In some embodiments, the modification is selected from the group consisting of: 2'OMe antisense oligonucleotide, 2'O-methoxyethyl antisense oligonucleotide, phosphorothioate antisense oligonucleotide, and LNA antisense oligonucleotide.

[0286] In some embodiments, the method of increasing the expression of UNC13A protein in a cell by at least 10% comprises contacting the cell with an antisense oligonucleotide, wherein the antisense oligonucleotide comprises 19 nucleotides complementary to the sequence shown in SEQ ID NO:1, and wherein the antisense oligonucleotide comprises the sequence shown in any of SEQ ID No:46, 61, 76, 91, 106, 121, 136, 151, 166, 181, 196, 211, 226, 241, 256, 271, 286, 301, 315, 316, 330, 331, 345, 346, 819, 830, 846, 856, 872, 883, 884, 892, 905, 1042 - 1058, and 1225 - 1247. In some embodiments, the antisense oligonucleotide consists of 19 nucleotides complementary to the sequence shown in SEQ ID NO:1, and wherein the antisense oligonucleotide consists of the sequence shown in any of SEQ ID No:46, 61, 76, 91, 106, 121, 136, 151, 166, 181, 196, 211, 226, 241, 256, 271, 286, 301, 315, 316, 330, 331, 345, 346, 819, 830, 846, 856, 872, 883, 884, 892, 905, 1042 - 1058, and 1225 - 1247. In some embodiments, wherein the antisense oligonucleotide is provided to the cell, the expression of UNC13A mRNA in the cell is increased to at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, or at least 50% of the level of UNC13A mRNA expressed in wild - type cells. In some embodiments, the increase in the expression of UNC13A mRNA in the cell is from about 10% to about 95%, 20% to about 85%, 30% to about 75%, 40% to about 65%, or 50% to about 55% of the level of UNC13A mRNA expressed in wild - type cells. In some embodiments, the increase in the expression of UNC13A mRNA in the cell is about 25% to about 1000%, about 50% to about 800%, about 100% to about 700%, about 200% to about 600%, or about 300% to about 500%. In some embodiments, wherein the antisense oligonucleotide is provided to the cell, the expression of UNC13A cryptic exon mRNA in the cell is reduced by about 10% to about 90%, about 20% to about 80%, about 30% to about 70%, or about 40% to about 60%. In some embodiments, wherein the antisense oligonucleotide is provided to the cell, the expression of UNC13A protein in the cell is increased to at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, or at least 50% of the level of UNC13A protein expressed in wild - type cells.In some embodiments, the increase in the expression of UNC13A protein in the cells is about 10% to about 95%, 20% to about 85%, 30% to about 75%, 40% to about 65%, or 50% to about 55% of the level of UNC13A protein expressed in wild-type cells. In some embodiments, the increase in the expression of UNC13A protein in the cells is about 25% to about 1000%, about 50% to about 800%, about 100% to about 700%, about 200% to about 600%, or about 300% to about 500%. In some embodiments, wherein an antisense oligonucleotide is provided to the cells, the expression of UNC13A protein is increased to a level sufficient to improve one or more neuronal functions selected from the group consisting of: neuronal signaling, neuronal morphology, neurite length, neurotransmitter release, action potential, neuronal synaptic plasticity, and neuronal lifespan. In some embodiments, the antisense oligonucleotide consists of the sequence shown in any one of SEQ ID NOs: 1059-1074, 1075-1089, 1090-1103, 1248-1270, 1271-1292, and 1293-1313. In some embodiments, the antisense oligonucleotide consists of the sequence shown in any one of SEQ ID NOs: 1059-1074, 1075-1089, and 1090-1103. In some embodiments, the antisense oligonucleotide consists of the sequence shown in any one of SEQ ID NOs: 1248-1270, 1271-1292, and 1293-1313. In some embodiments, the antisense oligonucleotide comprises a modification. In some embodiments, the modification is selected from the group consisting of: 2'-OMe antisense oligonucleotide, 2'-O-methoxyethyl antisense oligonucleotide, phosphorothioate antisense oligonucleotide, and LNA antisense oligonucleotide.

[0287] In some embodiments, the method of increasing the expression of UNC13A protein in a cell by at least 10% comprises contacting the cell with an antisense oligonucleotide, wherein the antisense oligonucleotide comprises 20 nucleotides complementary to the sequence shown in SEQ ID NO:1, and wherein the antisense oligonucleotide comprises the sequence shown in any of SEQ ID NO:47, 48, 62, 63, 77, 78, 92, 93, 107, 108, 122, 123, 137, 138, 152, 153, 167, 168, 182, 183, 197, 198, 212, 213, 227, 228, 242, 243, 257, 258, 272, 273, 287, 288, 302, 303, 317, 332, 347, 911, 936, 937, 1059 - 1074, and 1248 - 1270. In some embodiments, the antisense oligonucleotide consists of 20 nucleotides complementary to the sequence shown in SEQ ID NO:1, and wherein the antisense oligonucleotide consists of the sequence shown in any of SEQ ID NO:47, 48, 62, 63, 77, 78, 92, 93, 107, 108, 122, 123, 137, 138, 152, 153, 167, 168, 182, 183, 197, 198, 212, 213, 227, 228, 242, 243, 257, 258, 272, 273, 287, 288, 302, 303, 317, 332, 347, 911, 936, 937, 1059 - 1074, and 1248 - 1270. In some embodiments, wherein the antisense oligonucleotide is provided to the cell, the expression of UNC13A mRNA in the cell is increased to at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, or at least 50% of the level of UNC13A mRNA expressed in wild - type cells. In some embodiments, the increase in the expression of UNC13A mRNA in the cell is about 10% to about 95%, 20% to about 85%, 30% to about 75%, 40% to about 65%, or 50% to about 55% of the level of UNC13A mRNA expressed in wild - type cells. In some embodiments, the increase in the expression of UNC13A mRNA in the cell is about 25% to about 1000%, about 50% to about 800%, about 100% to about 700%, about 200% to about 600%, or about 300% to about 500%. In some embodiments, wherein the antisense oligonucleotide is provided to the cell, the expression of UNC13A cryptic exon mRNA in the cell is reduced by about 10% to about 90%, about 20% to about 80%, about 30% to about 70%, or about 40% to about 60%.In some embodiments, where an antisense oligonucleotide is provided to a cell, the expression of UNC13A protein in the cell is increased to at least 25%, at least 30%, at least 35%, at least 40%, at least 45% or at least 50% of the level of UNC13A protein expressed in wild-type cells. In some embodiments, the increase in the expression of UNC13A protein in the cell is from about 10% to about 95%, 20% to about 85%, 30% to about 75%, 40% to about 65% or 50% to about 55% of the level of UNC13A protein expressed in wild-type cells. In some embodiments, the increase in the expression of UNC13A protein in the cell is from about 25% to about 1000%, about 50% to about 800%, about 100% to about 700%, about 200% to about 600% or about 300% to about 500%. In some embodiments, where an antisense oligonucleotide is provided to a cell, the expression of UNC13A protein is increased to a level sufficient to improve the function of one or more neurons selected from the group consisting of: neuronal signaling, neuronal morphology, neurite length, neurotransmitter release, action potential, neuronal synaptic plasticity and neuronal lifespan. In some embodiments, the antisense oligonucleotide consists of a sequence shown in any one of SEQ ID NO: 1059 - 1074, 1075 - 1089, 1090 - 1103, 1248 - 1270, 1271 - 1292 and 1293 - 1313. In some embodiments, the antisense oligonucleotide consists of a sequence shown in any one of SEQ ID NO: 1059 - 1074, 1075 - 1089 and 1090 - 1103. In some embodiments, the antisense oligonucleotide consists of a sequence shown in any one of SEQ ID NO: 1248 - 1270, 1271 - 1292 and 1293 - 1313. In some embodiments, the antisense oligonucleotide comprises a modification. In some embodiments, the modification is selected from the group consisting of: 2'OMe antisense oligonucleotide, 2'O-methoxyethyl antisense oligonucleotide, phosphorothioate antisense oligonucleotide and LNA antisense oligonucleotide.

[0288] In some embodiments, the method of increasing the expression of UNC13A protein in a cell by at least 10% comprises contacting the cell with an antisense oligonucleotide, wherein the antisense oligonucleotide comprises 21 nucleotides complementary to the sequence shown in SEQ ID NO:1, and wherein the antisense oligonucleotide comprises any of the sequences shown in SEQ ID NOs: 20-37, 49, 64, 79, 94, 109, 124, 139, 154, 169, 184, 199, 214, 229, 244, 259, 274, 289, 304, 318, 319, 333, 334, 348, 349, 400-517, 1075-1089, and 1271-1292. In some embodiments, the antisense oligonucleotide consists of 21 nucleotides complementary to the sequence shown in SEQ ID NO:1, and wherein the antisense oligonucleotide consists of any of the sequences shown in SEQ ID NOs: 20-37, 49, 64, 79, 94, 109, 124, 139, 154, 169, 184, 199, 214, 229, 244, 259, 274, 289, 304, 318, 319, 333, 334, 348, 349, 400-517, 1075-1089, and 1271-1292. In some embodiments, wherein the antisense oligonucleotide is provided to the cell, the expression of UNC13A mRNA in the cell is increased to at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, or at least 50% of the level of UNC13A mRNA expressed in wild-type cells. In some embodiments, the increase in the expression of UNC13A mRNA in the cell is from about 10% to about 95%, 20% to about 85%, 30% to about 75%, 40% to about 65%, or 50% to about 55% of the level of UNC13A mRNA expressed in wild-type cells. In some embodiments, the increase in the expression of UNC13A mRNA in the cell is about 25% to about 1000%, about 50% to about 800%, about 100% to about 700%, about 200% to about 600%, or about 300% to about 500%. In some embodiments, wherein the antisense oligonucleotide is provided to the cell, the expression of UNC13A cryptic exon mRNA in the cell is decreased by about 10% to about 90%, about 20% to about 80%, about 30% to about 70%, or about 40% to about 60%. In some embodiments, wherein the antisense oligonucleotide is provided to the cell, the expression of UNC13A protein in the cell is increased to at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, or at least 50% of the level of UNC13A protein expressed in wild-type cells.In some embodiments, the increased expression of UNC13A protein in the cells is about 10% to about 95%, 20% to about 85%, 30% to about 75%, 40% to about 65%, or 50% to about 55% of the level of UNC13A protein expressed in wild-type cells. In some embodiments, the increased expression of UNC13A protein in the cells is about 25% to about 1000%, about 50% to about 800%, about 100% to about 700%, about 200% to about 600%, or about 300% to about 500%. In some embodiments, wherein an antisense oligonucleotide is provided to the cells, the expression of UNC13A protein is increased to a level sufficient to improve one or more neuronal functions selected from the group consisting of: neuronal signaling, neuronal morphology, neurite length, neurotransmitter release, action potential, neuronal synaptic plasticity, and neuronal lifespan. In some embodiments, the antisense oligonucleotide consists of the sequence shown in any one of SEQ ID NO: 1059-1074, 1075-1089, 1090-1103, 1248-1270, 1271-1292, and 1293-1313. In some embodiments, the antisense oligonucleotide consists of the sequence shown in any one of SEQ ID NO: 1059-1074, 1075-1089, and 1090-1103. In some embodiments, the antisense oligonucleotide consists of the sequence shown in any one of SEQ ID NO: 1248-1270, 1271-1292, and 1293-1313. In some embodiments, the antisense oligonucleotide comprises a modification. In some embodiments, the modification is selected from the group consisting of: 2'-OMe antisense oligonucleotide, 2'-O-methoxyethyl antisense oligonucleotide, phosphorothioate antisense oligonucleotide, and LNA antisense oligonucleotide.

[0289] In some embodiments, the method of increasing the expression of UNC13A protein in a cell by at least 10% comprises contacting the cell with an antisense oligonucleotide, wherein the antisense oligonucleotide comprises 22 nucleotides complementary to the sequence shown in SEQ ID NO:1, and wherein the antisense oligonucleotide comprises the sequence shown in any one of SEQ ID No:50, 51, 65, 66, 80, 81, 95, 96, 110, 111, 125, 126, 140, 141, 155, 156, 170, 171, 185, 186, 200, 201, 215, 216, 230, 231, 245, 246, 260, 261, 275, 276, 290, 291, 305, 306, 320, 335, 350, 912, 913, 914, 938, 939, 940, 941, 1090-1103 and 1293-1313. In some embodiments, the antisense oligonucleotide consists of 22 nucleotides complementary to the sequence shown in SEQ ID NO:1, and wherein the antisense oligonucleotide consists of the sequence shown in any one of SEQ ID NO:50, 51, 65, 66, 80, 81, 95, 96, 110, 111, 125, 126, 140, 141, 155, 156, 170, 171, 185, 186, 200, 201, 215, 216, 230, 231, 245, 246, 260, 261, 275, 276, 290, 291, 305, 306, 320, 335, 350, 912, 913, 914, 938, 939, 940, 941, 1090-1103 and 1293-1313. In some embodiments, wherein the antisense oligonucleotide is provided to the cell, the expression of UNC13A mRNA in the cell is increased to at least 25%, at least 30%, at least 35%, at least 40%, at least 45% or at least 50% of the level of UNC13A mRNA expressed in wild-type cells. In some embodiments, the increase in the expression of UNC13A mRNA in the cell is from about 10% to about 95%, 20% to about 85%, 30% to about 75%, 40% to about 65% or 50% to about 55% of the level of UNC13A mRNA expressed in wild-type cells. In some embodiments, the increase in the expression of UNC13A mRNA in the cell is about 25% to about 1000%, about 50% to about 800%, about 100% to about 700%, about 200% to about 600% or about 300% to about 500%. In some embodiments, wherein the antisense oligonucleotide is provided to the cell, the expression of UNC13A cryptic exon mRNA in the cell is decreased by about 10% to about 90%, about 20% to about 80%, about 30% to about 70% or about 40% to about 60%.In some embodiments, where an antisense oligonucleotide is provided to a cell, the expression of UNC13A protein in the cell is increased to at least 25%, at least 30%, at least 35%, at least 40%, at least 45% or at least 50% of the level of UNC13A protein expressed in wild-type cells. In some embodiments, the increase in the expression of UNC13A protein in the cell is from about 10% to about 95%, 20% to about 85%, 30% to about 75%, 40% to about 65% or 50% to about 55% of the level of UNC13A protein expressed in wild-type cells. In some embodiments, the increase in the expression of UNC13A protein in the cell is about 25% to about 1000%, about 50% to about 800%, about 100% to about 700%, about 200% to about 600% or about 300% to about 500%. In some embodiments, where an antisense oligonucleotide is provided to a cell, the expression of UNC13A protein is increased to a level sufficient to improve the function of one or more neurons selected from the group consisting of: neuronal signaling, neuronal morphology, neurite length, neurotransmitter release, action potential, neuronal synaptic plasticity, and neuronal lifespan. In some embodiments, the antisense oligonucleotide consists of the sequence shown in any one of SEQ ID NO: 1059-1074, 1075-1089, 1090-1103, 1248-1270, 1271-1292, and 1293-1313. In some embodiments, the antisense oligonucleotide consists of the sequence shown in any one of SEQ ID NO: 1059-1074, 1075-1089, and 1090-1103. In some embodiments, the antisense oligonucleotide consists of the sequence shown in any one of SEQ ID NO: 1248-1270, 1271-1292, and 1293-1313. In some embodiments, the a...

Claims

1. A method for increasing the expression of UNC13A protein in a cell by at least 10%, the method comprising contacting the cell with an antisense oligonucleotide comprising 13 to 30 nucleotides, wherein the antisense oligonucleotide is complementary to the sequence shown in SEQ ID NO:

1.

2. A method for increasing the expression of UNC13A protein in a cell by at least 10%, the method comprising contacting the cell with an antisense oligonucleotide comprising 13 to 30 nucleotides, wherein the antisense oligonucleotide is complementary to the sequence shown in any one of SEQ ID NO:1339 - 1342 or a part thereof.

3. The method according to claim 1 or claim 2, wherein the antisense oligonucleotide comprises the sequence shown in any one of SEQ ID NO:20 - 355, 400 - 638, 810 - 957 or 961 - 1332.

4. The method according to any one of claims 1 to 3, wherein the antisense oligonucleotide consists of 13 to 25 nucleotides, and wherein the antisense oligonucleotide is complementary to the sequence shown in SEQ ID NO:

1.

5. The method according to any one of claims 1 to 4, wherein the antisense oligonucleotide consists of 13, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24 or 25 nucleotides, and wherein the antisense oligonucleotide is complementary to the sequence shown in SEQ ID NO:

1.

6. The method according to any one of claims 1 to 5, wherein the antisense oligonucleotide consists of 19, 20, 21 or 22 nucleotides, and wherein the antisense oligonucleotide is complementary to the sequence shown in SEQ ID NO:

1.

7. The method according to any one of claims 1 to 5, wherein the antisense oligonucleotide consists of 13 nucleotides complementary to the sequence shown in SEQ ID NO:1, and wherein the antisense oligonucleotide consists of the sequence shown in any one of SEQ ID NO:961 - 983 and 1116 - 1145.

8. The method according to any one of claims 1 to 5, wherein the antisense oligonucleotide consists of 15 nucleotides complementary to the sequence shown in SEQ ID NO:1, and wherein the antisense oligonucleotide consists of the sequence shown in any one of SEQ ID NO:984 - 1004 and 1146 - 1173.

9. The method according to any one of claims 1 to 5, wherein the antisense oligonucleotide consists of 16 nucleotides complementary to the sequence shown in SEQ ID NO:1, and wherein the antisense oligonucleotide consists of the sequence shown in any one of SEQ ID NO:41, 42, 56, 57, 71, 72, 86, 87, 101, 102, 116, 117, 131, 132, 146, 147, 161, 162, 176, 177, 191, 192, 206, 207, 221, 222, 236, 237, 251, 252, 266, 267, 281, 282, 296, 297, 311, 326, 341, 810, 811, 812, 813, 826, 827, 835, 838, 841, 842, 851, 852, 861, 864, 867, 868, 877, 878, 895, 900, 901, 910, 934 and 935.

10. The method according to any one of claims 1 to 5, wherein the antisense oligonucleotide consists of 17 nucleotides complementary to the sequence shown in SEQ ID NO:1, and wherein the antisense oligonucleotide consists of the sequence shown in any one of SEQ ID NO:43, 58, 73, 88, 103, 118, 133, 148, 163, 178, 193, 208, 223, 238, 253, 268, 283, 298, 312, 313, 327, 328, 342, 343, 814, 815, 816, 828, 829, 836, 839, 843, 844, 853, 854, 862, 865, 869, 870, 879, 880, 881, 891, 896, 897, 902, 903, 1005 - 1023 and 1174 - 1199.

11. The method according to any one of claims 1 to 5, wherein the antisense oligonucleotide consists of 18 nucleotides complementary to the sequence shown in SEQ ID NO:1, and wherein the antisense oligonucleotide consists of the sequence shown in any one of SEQ ID NO:38 - 40, 44, 45, 59, 60, 74, 75, 89, 90, 104, 105, 119, 120, 134, 135, 149, 150, 164, 165, 179, 180, 194, 195, 209, 210, 224, 225, 239, 240, 254, 255, 269, 270, 284, 285, 299, 300, 314, 329, 344, 518 - 638, 817, 818, 845, 855, 871, 882, 904, 958, 959, 960, 1024 - 1041 and 1200 - 1224.

12. The method according to any one of claims 1 to 6, wherein the antisense oligonucleotide consists of 19 nucleotides complementary to the sequence shown in SEQ ID NO:1, and wherein the antisense oligonucleotide consists of the sequence shown in any one of SEQ ID No:46, 61, 76, 91, 106, 121, 136, 151, 166, 181, 196, 211, 226, 241, 256, 271, 286, 301, 315, 316, 330, 331, 345, 346, 819, 830, 846, 856, 872, 883, 884, 892, 905, 1042 - 1058 and 1225 - 1247.

13. The method according to any one of claims 1 to 6, wherein the antisense oligonucleotide consists of 20 nucleotides complementary to the sequence shown in SEQ ID NO:1, and wherein the antisense oligonucleotide consists of the sequence shown in any one of SEQ ID NO:47, 48, 62, 63, 77, 78, 92, 93, 107, 108, 122, 123, 137, 138, 152, 153, 167, 168, 182, 183, 197, 198, 212, 213, 227, 228, 242, 243, 257, 258, 272, 273, 287, 288, 302, 303, 317, 332, 347, 911, 936, 937, 1059 - 1074 and 1248 - 1270.

14. The method according to any one of claims 1 to 6, wherein the antisense oligonucleotide consists of 21 nucleotides complementary to the sequence shown in SEQ ID NO:1, and wherein the antisense oligonucleotide consists of the sequence shown in any one of SEQ ID NO:20 - 37, 49, 64, 79, 94, 109, 124, 139, 154, 169, 184, 199, 214, 229, 244, 259, 274, 289, 304, 318, 319, 333, 334, 348, 349, 400 - 517, 1075 - 1089 and 1271 - 1292.

15. The method according to any one of claims 1 to 6, wherein the antisense oligonucleotide consists of 22 nucleotides complementary to the sequence shown in SEQ ID NO:1, and wherein the antisense oligonucleotide consists of the sequence shown in any one of SEQ ID NO:50, 51, 65, 66, 80, 81, 95, 96, 110, 111, 125, 126, 140, 141, 155, 156, 170, 171, 185, 186, 200, 201, 215, 216, 230, 231, 245, 246, 260, 261, 275, 276, 290, 291, 305, 306, 320, 335, 350, 912, 913, 914, 938, 939, 940, 941, 1090 - 1103 and 1293 - 1313.

16. The method according to any one of claims 1 to 5, wherein the antisense oligonucleotide consists of 23 nucleotides complementary to the sequence shown in SEQ ID NO:1, and wherein the antisense oligonucleotide consists of the sequence shown in any one of SEQ ID No:52, 67, 82, 97, 112, 127, 142, 157, 172, 187, 202, 217, 232, 247, 262, 277, 292, 307, 321, 322, 336, 337, 351, 352, 820, 831, 847, 857, 873, 885, 886, 893, 906, 915 - 918, 930 and 942 - 945.

17. The method according to any one of claims 1 to 5, wherein the antisense oligonucleotide consists of 24 nucleotides complementary to the sequence shown in SEQ ID NO:1, and wherein the antisense oligonucleotide consists of the sequence shown in any one of SEQ ID NO:53, 54, 68, 69, 83, 84, 98, 99, 113, 114, 128, 129, 143, 144, 158, 159, 173, 174, 188, 189, 203, 204, 218, 219, 233, 234, 248, 249, 263, 264, 278, 279, 293, 294, 308, 309, 323, 338, 353, 821, 822, 832, 848, 858, 874, 887, 907, 919, 920 - 923, 931, 946 - 951, 1104 - 1115 and 1314 - 1332.

18. The method according to any one of claims 1 to 5, wherein the antisense oligonucleotide consists of 25 nucleotides complementary to the sequence shown in SEQ ID NO:1, and wherein the antisense oligonucleotide consists of the sequence shown in any one of SEQ ID No:55, 70, 85, 100, 115, 130, 145, 160, 175, 190, 205, 220, 235, 250, 265, 280, 295, 310, 324, 325, 339, 340, 354, 355, 823, 824, 825, 833, 834, 837, 840, 849, 850, 859, 860, 863, 866, 875, 876, 888 - 890, 894, 898, 899, 908, 909, 924 - 929, 932, 933, 952 and 953 - 957.

19. The method according to any one of claims 1 to 18, wherein the antisense oligonucleotide is complementary to the sequence shown in any one of SEQ ID NO:700 - 703.

20. The method according to any one of claims 1 to 19, wherein the antisense oligonucleotide has at least 80%, at least 90%, at least 95%, at least 98% or 100% complementarity with the sequence shown in any one of SEQ ID NO:700 - 703.

21. The method according to any one of claims 1 to 20, wherein the antisense oligonucleotide has at least 80%, at least 90%, at least 95%, at least 98% or 100% complementarity with the sequence shown in SEQ ID NO:

701.

22. The method according to any one of claims 1 to 21, wherein the antisense oligonucleotide has at least 80%, at least 90%, at least 95%, at least 98% or 100% complementarity with the sequence shown in SEQ ID NO:

703.

23. The method according to any one of claims 1 to 22, wherein the antisense oligonucleotide is complementary to the sequence shown in any one of SEQ ID NO:1339 - 1342 or a part thereof.

24. The method according to any one of claims 1 to 21 and 23, wherein the antisense oligonucleotide is complementary to the sequence shown in SEQ IDNO:1339 or a part thereof.

25. The method according to any one of claims 1 to 21 and 23 to 24, wherein the antisense oligonucleotide is complementary to the sequence shown in SEQID NO:1340 or a part thereof.

26. The method according to any one of claims 1 to 20 and 22 to 23, wherein the antisense oligonucleotide is complementary to the sequence shown in SEQID NO:1341 or a part thereof.

27. The method according to any one of claims 1 to 20, 22 to 23 and 26, wherein the antisense oligonucleotide is complementary to the sequence shown in SEQ ID NO:1342 or a part thereof.

28. The method according to any one of claims 1 to 27, wherein the antisense oligonucleotide is complementary to at least 1, at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, at least 11, at least 12, or at least 13 nucleotides of the sequence shown in any one of SEQ ID NOs: 1339 - 1342.

29. The method according to any one of claims 1 to 28, wherein the antisense oligonucleotide is complementary to at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, at least 11, at least 12, or at least 13 consecutive nucleotides of the sequence shown in any one of SEQ ID NOs: 1339 - 1342.

30. The method according to claim 29, wherein the antisense oligonucleotide is at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99%, or 100% complementary to the sequence shown in SEQ ID NO: 1339.

31. The method according to claim 29, wherein the antisense oligonucleotide is at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99%, or 100% complementary to the sequence shown in SEQ ID NO: 1340.

32. The method according to claim 29, wherein the antisense oligonucleotide is at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99%, or 100% complementary to the sequence shown in SEQ ID NO: 1341.

33. The method according to claim 29, wherein the antisense oligonucleotide is at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99%, or 100% complementary to the sequence shown in SEQ ID NO: 1342.

34. The method according to any one of claims 1 to 33, wherein the cell expresses a low level of UNC13A before contacting the cell with the antisense oligonucleotide.

35. The method according to any one of claims 1 to 34, further comprising measuring the level of UNC13A protein in the cell.

36. The method according to claim 35, wherein the level of UNC13A protein is measured by Western blot, imaging, ELISA, fluorescence reporter assay, luminescence assay, or immunohistochemistry.

37. The method according to any one of claims 1 to 36, wherein the secondary structure of UNC13A mRNA is altered by the binding of the antisense oligonucleotide to the UNC13A transcript.

38. The method according to any one of claims 1 to 37, wherein the antisense oligonucleotide sterically blocks the inclusion of the UNC13A cryptic exon.

39. The method according to any one of claims 1 to 38, wherein the antisense oligonucleotide has at least 80%, at least 90%, at least 95%, at least 98% or 100% complementarity with the sequence shown in SEQ ID NO:

1.

40. The method according to any one of claims 1 to 39, wherein the antisense oligonucleotide restores the expression of UNC13A mRNA in the cell to at least 25%, at least 30%, at least 35%, at least 40%, at least 45% or at least 50% of the level of UNC13A mRNA expressed in wild-type cells.

41. The method according to any one of claims 1 to 40, wherein the antisense oligonucleotide increases the expression of UNC13A mRNA in the cell by at least 50%, at least 100%, at least 200%, at least 300%, at least 400% or at least 500%.

42. The method according to claim 40 or claim 41, wherein the antisense oligonucleotide consists of the sequence shown in any one of SEQ ID NO: 1059 - 1074, 1075 - 1089, 1090 - 1103, 1248 - 1270, 1271 - 1292 and 1293 - 1313.

43. The method according to any one of claims 1 to 42, wherein the antisense oligonucleotide reduces the expression of UNC13A cryptic exon splice variant mRNA by at least 10%.

44. The method according to any one of claims 1 to 43, wherein the antisense oligonucleotide reduces the expression of UNC13A cryptic exon mRNA by at least 10%, at least 20%, at least 30%, at least 40% or at least 50% of the level of UNC13A cryptic exon mRNA expressed in cells treated with shRNA, miRNA, antisense oligonucleotide targeting TARDBP.

45. The method according to any one of claims 1 to 44, wherein the antisense oligonucleotide reduces the expression of UNC13A cryptic exon mRNA in the cell by at least 10%, at least 20%, at least 30%, at least 40% or at least 50%.

46. The method according to claim 44 or claim 45, wherein the antisense oligonucleotide consists of the sequence shown in any one of SEQ ID NO: 1059 - 1074, 1075 - 1089, 1090 - 1103, 1248 - 1270, 1271 - 1292 and 1293 - 1313.

47. The method according to any one of claims 38 to 46, wherein the UNC13A cryptic exon splice variant contains the sequence shown in SEQ ID NO: 7 or SEQ ID NO:

9.

48. The method according to any one of claims 38 to 47, wherein after contacting the cell with the antisense oligonucleotide, the ratio of UNC13A transcripts not containing the cryptic exon splice variant to UNC13A transcripts containing the cryptic exon splice variant is at least 10:

1.

49. The method according to any one of claims 38 to 49, wherein the presence and / or amount of the cryptic exon splicing variant is measured using qPCR.

50. The method according to any one of claims 1 to 49, wherein the cell is a neuron.

51. The method according to any one of claims 1 to 50, wherein contacting the cell with the antisense oligonucleotide increases the expression of the UNC13A protein to a level sufficient to improve one or more neuronal functions.

52. The method according to any one of claims 1 to 51, wherein the antisense oligonucleotide restores the expression of the UNC13A protein in the cell to at least 10%, at least 20%, at least 30%, at least 40% or at least 50% of the level of the UNC13A protein expressed in wild-type cells.

53. The method according to any one of claims 1 to 52, wherein the antisense oligonucleotide increases the expression of the UNC13A protein in the cell by at least 50%, at least 100%, at least 200%, at least 300%, at least 400% or at least 500%.

54. The method according to claim 52 or claim 53, wherein the antisense oligonucleotide consists of the sequence shown in any one of SEQ ID NO: 1059 - 1074, 1075 - 1089, 1090 - 1103, 1248 - 1270, 1271 - 1292 and 1293 - 1313.

55. The method according to any one of claims 1 to 54, wherein the antisense oligonucleotide increases the expression of UNC13A mRNA in the cell by at least 50%, decreases the expression of UNC13A cryptic exon mRNA in the cell by at least 25%, and increases the expression of UNC13A protein in the cell by at least 50%.

56. The method according to any one of claims 1 to 6, 13 to 15 and 19 to 55, wherein the antisense oligonucleotide consists of the sequence shown in any one of SEQ ID NO: 1059 - 1074, 1075 - 1089, 1090 - 1103, 1248 - 1270, 1271 - 1292 and 1293 - 1313.

57. The method according to any one of claims 1 to 6, 13 to 15 and 19 to 56, wherein the antisense oligonucleotide consists of the sequence shown in any one of SEQ ID NO: 1059 - 1074, 1075 - 1089 and 1090 - 1103.

58. The method according to any one of claims 1 to 6, 13 to 15 and 19 to 56, wherein the antisense oligonucleotide consists of the sequence shown in any one of SEQ ID NO: 1248 - 1270, 1271 - 1292 and 1293 - 1313.

59. The method according to any one of claims 50 to 58, wherein the neuron is a motor neuron, a cortical neuron, an interneuron or an excitatory neuron.

60. The method according to any one of claims 51 to 59, wherein the one or more neuronal functions are selected from the group consisting of: neuronal signaling, neuronal morphology, neurite length, neurotransmitter release, action potential, neuronal synaptic plasticity, and neuronal lifespan.

61. The method according to any one of claims 1 to 60, wherein the cell is heterozygous or homozygous for the risk allele.

62. The method according to claim 61, wherein the risk allele is selected from the group consisting of: rs12608932 (hg38 chr19:17,641,880A→C), rs12973192 (hg38 chr19:17,642,430C→G), rs56041637 (hg38 chr19:17,642,033 - 17,642,056 0 - 2CATC repeat→3 - 5CATC repeat), and rs62121687 (hg38 chr19:17,642,351C→A) or any combination thereof.

63. The method according to any one of claims 1 to 62, wherein the cell comprises a mutation associated with amyotrophic lateral sclerosis (ALS) or frontotemporal dementia (FTD).

64. The method according to any one of claims 1 to 63, wherein the antisense oligonucleotide is provided to the cell by transfection.

65. The method according to any one of claims 1 to 64, wherein the cell expresses a low level of the TDP - 43 protein.

66. The method according to any one of claims 1 to 65, wherein the cell is a human cell or a murine cell.

67. The method according to any one of claims 1 to 66, wherein the antisense oligonucleotide does not cause any change in cell morphology.

68. The method according to any one of claims 1 to 67, wherein the antisense oligonucleotide does not cause neuronal toxicity.

69. The method according to any one of claims 1 to 68, wherein the antisense oligonucleotide does not bind to genomic sequences other than the sequence shown in SEQ ID NO:

1.

70. A method of treating amyotrophic lateral sclerosis (ALS) or frontotemporal dementia (FTD) in an individual, the method comprising administering to the individual an antisense oligonucleotide comprising 13 to 30 nucleotides, wherein the antisense oligonucleotide is complementary to the sequence shown in SEQ ID NO:1, and wherein the individual is homozygous for the reference allele, homozygous for the risk allele, heterozygous for the risk allele, or comprises a mutation associated with ALS or FTD.

71. The method according to claim 70, wherein the risk allele is selected from the group consisting of: rs12608932 (hg38 chr19:17,641,880A→C), rs12973192 (hg38 chr19:17,642,430C→G), rs56041637 (hg38 chr19:17,642,033-17,642,056 0-2CATC repeat→3-5CATC repeat), and rs62121687 (hg38 chr19:17,642,351C→A) or any combination thereof.

72. The method according to claim 70, wherein the mutation associated with ALS or FTD is a mutation in TDP43.

73. The method according to any one of claims 1 to 72, wherein the antisense oligonucleotide has a sequence complementary to the sequence of UNC13A mRNA, the sequence of UNC13A mRNA being between the 5' splice donor site and the 3' splice acceptor site of a cryptic exon located between exons 20 and 21 of the UNC13A gene.

74. The method according to any one of claims 1 to 73, wherein the antisense oligonucleotide comprises a 2'-OMe antisense oligonucleotide, a 2'-O-methoxyethyl antisense oligonucleotide, a phosphorothioate antisense oligonucleotide, or a locked nucleic acid (LNA) antisense oligonucleotide.

75. An antisense oligonucleotide for regulating UNC13A splicing, wherein the antisense oligonucleotide has a sequence complementary to the sequence of UNC13A mRNA, the sequence of UNC13A mRNA being between the 5' splice donor site and the 3' splice acceptor site of a cryptic exon located between exons 20 and 21 of the UNC13A gene.

76. An antisense oligonucleotide for regulating UNC13A splicing and comprising 13 to 30 nucleotides, wherein the antisense oligonucleotide comprises the sequence shown in any one of SEQ ID NOs: 20-355, 400-638, 810-959, or 961-1332.

77. The antisense oligonucleotide according to claim 75 or claim 76, wherein the antisense oligonucleotide is complementary to the sequence shown in SEQ ID NO:

1.

78. The antisense oligonucleotide according to any one of claims 75 to 77, wherein the antisense oligonucleotide is complementary to the sequence shown in any one of SEQ ID NOs: 700-703.

79. The antisense oligonucleotide according to any one of claims 75 to 78, which comprises at least 80%, at least 90%, at least 95%, at least 98%, or 100% complementarity to the sequence shown in any one of SEQ ID NOs: 700-703.

80. The antisense oligonucleotide according to any one of claims 75 to 79, which comprises at least 80%, at least 90%, at least 95%, at least 98% or 100% complementarity to the sequence shown in SEQ ID NO:

701.

81. The antisense oligonucleotide according to any one of claims 75 to 79, which comprises at least 80%, at least 90%, at least 95%, at least 98% or 100% complementarity to the sequence shown in SEQ ID NO:

703.

82. The antisense oligonucleotide according to any one of claims 75 to 81, wherein the antisense oligonucleotide is complementary to the sequence or a part thereof shown in any one of SEQ ID NOs: 1339 - 1342.

83. The antisense oligonucleotide according to any one of claims 75 to 80 and 82 to 83, wherein the antisense oligonucleotide is complementary to the sequence or a part thereof shown in SEQ ID NO: 1339.

84. The antisense oligonucleotide according to any one of claims 75 to 80 and 82 to 84, wherein the antisense oligonucleotide is complementary to the sequence or a part thereof shown in SEQ ID NO: 1340.

85. The antisense oligonucleotide according to any one of claims 75 to 79 and 81 to 82, wherein the antisense oligonucleotide is complementary to the sequence or a part thereof shown in SEQ ID NO: 1341.

86. The antisense oligonucleotide according to any one of claims 75 to 79, 81 to 82 and 85, wherein the antisense oligonucleotide is complementary to the sequence or a part thereof shown in SEQ ID NO: 1342.

87. The antisense oligonucleotide according to any one of claims 75 to 86, wherein the antisense oligonucleotide is complementary to at least 1, at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, at least 11, at least 12 or at least 13 nucleotides of the sequence shown in any one of SEQ ID NOs: 1339 - 1342.

88. The antisense oligonucleotide according to any one of claims 75 to 87, wherein the antisense oligonucleotide is complementary to at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, at least 11, at least 12 or at least 13 consecutive nucleotides of the sequence shown in any one of SEQ ID NOs: 1339 - 1342.

89. The antisense oligonucleotide according to claim 88, wherein the antisense oligonucleotide is at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% complementary to the sequence shown in SEQ ID NO: 1339.

90. The antisense oligonucleotide according to claim 88, wherein the antisense oligonucleotide is at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% complementary to the sequence shown in SEQ ID NO: 1340.

91. The antisense oligonucleotide according to claim 88, wherein the antisense oligonucleotide is at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% complementary to the sequence shown in SEQ ID NO: 1341.

92. The antisense oligonucleotide according to claim 88, wherein the antisense oligonucleotide is at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100% complementary to the sequence shown in SEQ ID NO: 1342.

93. An antisense oligonucleotide comprising the sequence shown in any one of SEQ ID NO: 20-355, 400-638, 810-959 or 961-1332.

94. The antisense oligonucleotide according to any one of claims 75 to 93, wherein the antisense oligonucleotide consists of 13-30 nucleotides.

95. The antisense oligonucleotide according to any one of claims 75 to 94, wherein the antisense oligonucleotide consists of 13-25 nucleotides.

96. The antisense oligonucleotide according to any one of claims 75 to 95, wherein the antisense oligonucleotide consists of 13, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24 or 25 nucleotides.

97. The antisense oligonucleotide according to any one of claims 75 to 96, wherein the antisense oligonucleotide consists of 19, 20, 21 or 22 nucleotides.

98. The antisense oligonucleotide according to any one of claims 75 to 97, wherein the antisense oligonucleotide is complementary to the sequence shown in SEQ ID NO:

1.

99. An antisense oligonucleotide consisting of the sequence shown in any one of SEQ ID NO: 20-355, 400-638, 810-959 or 961-1332.

100. An antisense oligonucleotide, wherein the antisense oligonucleotide is complementary to the sequence shown in any one of SEQ ID NO: 1339-1342 or a part thereof.

101. The antisense oligonucleotide according to any one of claims 75 to 96 and 98 to 100, wherein the antisense oligonucleotide consists of 13 nucleotides complementary to the sequence shown in SEQ ID NO: 1, and wherein the antisense oligonucleotide consists of the sequence shown in any one of SEQ ID NO: 961-983 and 1116-1145.

102. An antisense oligonucleotide according to any one of claims 75 to 96 and 98 to 100, wherein the antisense oligonucleotide consists of 15 nucleotides complementary to the sequence shown in SEQ ID NO: 1, and wherein the antisense oligonucleotide consists of the sequence shown in any one of SEQ ID NOs: 984-1004 and 1146-1173.

103. An antisense oligonucleotide according to any one of claims 75 to 96 and 98 to 100, wherein the antisense oligonucleotide consists of 16 nucleotides complementary to the sequence shown in SEQ ID NO: 1, and wherein the antisense oligonucleotide consists of the sequence shown in any one of SEQ ID NOs: 41, 42, 56, 57, 71, 72, 86, 87, 101, 102, 116, 117, 131, 132, 146, 147, 161, 162, 176, 177, 191, 192, 206, 207, 221, 222, 236, 237, 251, 252, 266, 267, 281, 282, 296, 297, 311, 326, 341, 810, 811, 812, 813, 826, 827, 835, 838, 841, 842, 851, 852, 861, 864, 867, 868, 877, 878, 895, 900, 901, 910, 934 and 935.

104. An antisense oligonucleotide according to any one of claims 75 to 96 and 98 to 100, wherein the antisense oligonucleotide consists of 17 nucleotides complementary to the sequence shown in SEQ ID NO: 1, and wherein the antisense oligonucleotide consists of the sequence shown in any one of SEQ ID NOs: 43, 58, 73, 88, 103, 118, 133, 148, 163, 178, 193, 208, 223, 238, 253, 268, 283, 298, 312, 313, 327, 328, 342, 343, 814, 815, 816, 828, 829, 836, 839, 843, 844, 853, 854, 862, 865, 869, 870, 879, 880, 881, 891, 896, 897, 902, 903, 1005-1023 and 1174-1199.

105. An antisense oligonucleotide according to any one of claims 75 to 96 and 98 to 100, wherein the antisense oligonucleotide consists of 18 nucleotides complementary to the sequence shown in SEQ ID NO: 1, and wherein the antisense oligonucleotide consists of the sequence shown in any one of SEQ ID NOs: 38 - 40, 44, 45, 59, 60, 74, 75, 89, 90, 104, 105, 119, 120, 134, 135, 149, 150, 164, 165, 179, 180, 194, 195, 209, 210, 224, 225, 239, 240, 254, 255, 269, 270, 284, 285, 299, 300, 314, 329, 344, 518 - 638, 817, 818, 845, 855, 871, 882, 904, 958, 959, 960, 1024 - 1041, and 1200 - 1224.

106. An antisense oligonucleotide according to any one of claims 75 to 100, wherein the antisense oligonucleotide consists of 19 nucleotides complementary to the sequence shown in SEQ ID NO: 1, and wherein the antisense oligonucleotide consists of the sequence shown in any one of SEQ ID NOs: 46, 61, 76, 91, 106, 121, 136, 151, 166, 181, 196, 211, 226, 241, 256, 271, 286, 301, 315, 316, 330, 331, 345, 346, 819, 830, 846, 856, 872, 883, 884, 892, 905, 1042 - 1058, and 1225 - 1247.

107. An antisense oligonucleotide according to any one of claims 75 to 100, wherein the antisense oligonucleotide consists of 20 nucleotides complementary to the sequence shown in SEQ ID NO: 1, and wherein the antisense oligonucleotide consists of the sequence shown in any one of SEQ ID NOs: 47, 48, 62, 63, 77, 78, 92, 93, 107, 108, 122, 123, 137, 138, 152, 153, 167, 168, 182, 183, 197, 198, 212, 213, 227, 228, 242, 243, 257, 258, 272, 273, 287, 288, 302, 303, 317, 332, 347, 911, 936, 937, 1059 - 1074, and 1248 - 1270.

108. The antisense oligonucleotide according to any one of claims 75 to 100, wherein the antisense oligonucleotide consists of 21 nucleotides complementary to the sequence shown in SEQ ID NO: 1, and wherein the antisense oligonucleotide consists of the sequence shown in any one of SEQ ID NOs: 20 - 37, 49, 64, 79, 94, 109, 124, 139, 154, 169, 184, 199, 214, 229, 244, 259, 274, 289, 304, 318, 319, 333, 334, 348, 349, 400 - 517, 1075 - 1089, and 1271 - 1292.

109. The antisense oligonucleotide according to any one of claims 75 to 100, wherein the antisense oligonucleotide consists of 22 nucleotides complementary to the sequence shown in SEQ ID NO: 1, and wherein the antisense oligonucleotide consists of the sequence shown in any one of SEQ ID NOs: 50, 51, 65, 66, 80, 81, 95, 96, 110, 111, 125, 126, 140, 141, 155, 156, 170, 171, 185, 186, 200, 201, 215, 216, 230, 231, 245, 246, 260, 261, 275, 276, 290, 291, 305, 306, 320, 335, 350, 912, 913, 914, 938, 939, 940, 941, 1090 - 1103, and 1293 - 1313.

110. The antisense oligonucleotide according to any one of claims 75 to 96 and 98 to 100, wherein the antisense oligonucleotide consists of 23 nucleotides complementary to the sequence shown in SEQ ID NO: 1, and wherein the antisense oligonucleotide consists of the sequence shown in any one of SEQ ID NOs: 52, 67, 82, 97, 112, 127, 142, 157, 172, 187, 202, 217, 232, 247, 262, 277, 292, 307, 321, 322, 336, 337, 351, 352, 820, 831, 847, 857, 873, 885, 886, 893, 906, 915 - 918, 930, and 942 - 945.

111. An antisense oligonucleotide according to any one of claims 75 to 96 and 98 to 100, wherein the antisense oligonucleotide consists of 24 nucleotides complementary to the sequence shown in SEQ ID NO: 1, and wherein the antisense oligonucleotide consists of the sequence shown in any one of SEQ ID NO: 53, 54, 68, 69, 83, 84, 98, 99, 113, 114, 128, 129, 143, 144, 158, 159, 173, 174, 188, 189, 203, 204, 218, 219, 233, 234, 248, 249, 263, 264, 278, 279, 293, 294, 308, 309, 323, 338, 353, 821, 822, 832, 848, 858, 874, 887, 907, 919, 920 - 923, 931, 946 - 951, 1104 - 1115 and 1314 - 1332.

112. An antisense oligonucleotide according to any one of claims 75 to 96 and 98 to 100, wherein the antisense oligonucleotide consists of 25 nucleotides complementary to the sequence shown in SEQ ID NO: 1, and wherein the antisense oligonucleotide consists of the sequence shown in any one of SEQ ID No: 55, 70, 85, 100, 115, 130, 145, 160, 175, 190, 205, 220, 235, 250, 265, 280, 295, 310, 324, 325, 339, 340, 354, 355, 823, 824, 825, 833, 834, 837, 840, 849, 850, 859, 860, 863, 866, 875, 876, 888 - 890, 894, 898, 899, 908, 909, 924 - 929, 932, 933, 952 and 953 - 957.

113. An antisense oligonucleotide according to any one of claims 75 to 80 and 82 to 112, wherein the antisense oligonucleotide is complementary to the sequence shown in SEQ ID NO: 1339 or a part thereof.

114. An antisense oligonucleotide according to any one of claims 75 to 80 and 82 to 113, wherein the antisense oligonucleotide is complementary to the sequence shown in SEQ ID NO: 1340 or a part thereof.

115. An antisense oligonucleotide according to any one of claims 75 to 79 and 81 to 112, wherein the antisense oligonucleotide is complementary to the sequence shown in SEQ ID NO: 1341 or a part thereof.

116. An antisense oligonucleotide according to any one of claims 75 to 79, 81 to 112 and 115, wherein the antisense oligonucleotide is complementary to the sequence shown in SEQ ID NO: 1342 or a part thereof.

117. The antisense oligonucleotide according to any one of claims 75 to 116, wherein the antisense oligonucleotide is complementary to at least 1, at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, at least 11, at least 12, or at least 13 nucleotides of the sequence shown in any one of SEQ ID NOs: 1339 - 1342.

118. The antisense oligonucleotide according to any one of claims 75 to 117, wherein the antisense oligonucleotide is complementary to at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, at least 11, at least 12, or at least 13 consecutive nucleotides of the sequence shown in any one of SEQ ID NOs: 1339 - 1342.

119. The antisense oligonucleotide according to claim 118, wherein the antisense oligonucleotide is at least 80%, 85%, at least 90%, at least 95%, at least 98%, at least 99%, or 100% complementary to the sequence shown in SEQ ID NO: 1339.

120. The antisense oligonucleotide according to claim 118, wherein the antisense oligonucleotide is at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99%, or 100% complementary to the sequence shown in SEQ ID NO: 1340.

121. The antisense oligonucleotide according to claim 118, wherein the antisense oligonucleotide is at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99%, or 100% complementary to the sequence shown in SEQ ID NO: 1341.

122. The antisense oligonucleotide according to claim 118, wherein the antisense oligonucleotide is at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99%, or 100% complementary to the sequence shown in SEQ ID NO: 1342.

123. The antisense oligonucleotide according to any one of claims 75 to 100, 107 to 109, and 113 to 122, wherein the antisense oligonucleotide consists of the sequence shown in any one of SEQ ID NOs: 1059 - 1074, 1075 - 1089, 1090 - 1103, 1248 - 1270, 1271 - 1292, and 1293 - 1313.

124. The antisense oligonucleotide according to any one of claims 75 to 123, wherein the antisense oligonucleotide contains a modification.

125. The antisense oligonucleotide according to any one of claims 75 to 124, wherein the 5'-end and / or 3'-end of the antisense oligonucleotide contains a modification.

126. An antisense oligonucleotide according to any one of claims 75 to 125, wherein the 5' and 3' ends of the antisense oligonucleotide comprise modifications, wherein 2, 3, 4, or 5 nucleotides are modified at each of the 5' and 3' ends of the antisense oligonucleotide.

127. An antisense oligonucleotide according to any one of claims 75 to 126, wherein at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 95%, or at least 98% of the nucleotides comprise modifications.

128. An antisense oligonucleotide according to any one of claims 75 to 127, wherein the modifications are selected from the group consisting of: 2'-OMe antisense oligonucleotides, 2'-O-methoxyethyl antisense oligonucleotides, phosphorothioate antisense oligonucleotides, and locked nucleic acid (LNA) antisense oligonucleotides.

129. An antisense oligonucleotide according to any one of claims 75 to 128, wherein the modification is a 2'-OMe antisense oligonucleotide.

130. A pharmaceutical composition comprising an antisense oligonucleotide according to any one of claims 75 to 129.

131. The pharmaceutical composition according to claim 130, which comprises an excipient and / or a buffer.

132. A kit comprising an antisense oligonucleotide according to any one of claims 75 to 129 or a pharmaceutical composition according to claim 130 or claim 131.

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