UNC13A antisense oligonucleotides and uses thereof

Abstract

The present disclosure, in some aspects, relates to compositions of UNCI3A antisense oligonucleotides (ASOs) and methods of using ASOs to alter UNCI3A cryptic exon splicing and increase UNCI3A protein expression. Such ASOs can be used to treat neurodegenerative disorders, particularly those associated with TAR-DNA binding protein-43 (TDP-43) pathologies, such as amyotrophic lateral sclerosis (ALS) or frontotemporal dementia (FTD).

Description

[Technical Field]

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS This application claims priority from U.S. Provisional Application No. 63 / 378,473, filed October 5, 2022, the contents of which are incorporated by reference in their entirety.

[0002] Submission of sequence listing as an ASCII text file The contents of the electronic sequence listing (792752001540seqlist.xml; size: 1,083,815 bytes, and creation date: October 4, 2023) are incorporated herein by reference in their entirety.

[0003] The present disclosure relates, in some aspects, to compositions and methods using antisense oligonucleotides targeted to the UNC13A cryptic exon locus to increase UNC13A protein expression. [Background technology]

[0004] A characteristic pathological feature of the neurodegenerative diseases amyotrophic lateral sclerosis (ALS) and frontotemporal dementia (FTD) is the depletion of the RNA-binding protein TDP-43 from neuronal nuclei in the brain and spinal cord. TDP-43, encoded by TARDBP, is an abundant and ubiquitously expressed RNA-binding protein normally localized in the nucleus. It plays a role in fundamental 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). Depletion of TDP-43 from an otherwise normal adult nervous system alters the splicing or expression levels of over 1,500 RNAs, including long intron-containing transcripts (2). The primary splicing regulatory function of TDP-43 is to suppress the inclusion of cryptic exons during splicing (4-7). Unlike normal conserved exons, these cryptic exons are hidden within introns and are normally excluded from mature mRNAs. When TDP-43 is depleted from cells, these hidden exons are spliced ​​into messenger RNA, often resulting in frameshifts, premature termination, and even nonsense-mediated decay of mRNA. However, the hidden splicing events that are key to disease remain to be identified. Therefore, there is a need to discover hidden splicing targets that are regulated by TDP-43 and also play a role in the pathogenesis of TDP-43 protein abnormalities as therapeutic targets. Methods and compositions that address these and other needs are provided herein. Summary of the Invention [Means for solving the problem]

[0005] Provided herein are antisense oligonucleotides (ASOs) that modulate UNC13A splicing, wherein the ASO sequence is complementary to a 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. The ASOs provided herein are 13 to 30 nucleotides in length. Also provided are modified ASOs containing one or more modified nucleotides, 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 herein. In some embodiments, provided herein are methods for increasing the expression of full-length UNC13A protein in cells by at least 10%. In other embodiments, provided herein are methods for treating a clinical disease in an individual, such as amyotrophic lateral sclerosis (ALS) or frontotemporal dementia (FTD). Also provided herein are pharmaceutical compositions and kits that include one or more of the ASOs described herein.

[0006] Provided herein is a method for increasing 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 that are complementary to the sequence set forth in SEQ ID NO:1.

[0007] Provided herein is a method for increasing expression of UNC13A protein in a cell by at least 10%, comprising contacting the cell with an antisense oligonucleotide comprising 13 to 30 nucleotides, wherein the antisense oligonucleotide is complementary to the sequence set forth in SEQ ID NO: 1. Also provided herein is a method for increasing expression of UNC13A protein in a cell by at least 10%, comprising contacting the cell with an antisense oligonucleotide comprising 13 to 30 nucleotides, wherein the antisense oligonucleotide is complementary to the sequence set forth in any one of SEQ ID NOs: 1339 to 1342 or a portion thereof. In some embodiments, the antisense oligonucleotide comprises the sequence set forth in any one of SEQ ID NOs: 20 to 355, 400 to 638, 810 to 957, or 961 to 1332.

[0008] In some embodiments, the antisense oligonucleotide consists of 13 to 25 nucleotides, and the antisense oligonucleotide is complementary to the sequence set forth 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, and the antisense oligonucleotide is complementary to the sequence set forth in SEQ ID NO: 1. In some embodiments, the antisense oligonucleotide consists of 19, 20, 21, or 22 nucleotides, and the antisense oligonucleotide is complementary to the sequence set forth in SEQ ID NO: 1.

[0009] In some embodiments, the antisense oligonucleotide consists of 13 nucleotides complementary to the sequence set forth in SEQ ID NO:1, wherein the antisense oligonucleotide consists of the sequence set forth 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 set forth in SEQ ID NO:1, wherein the antisense oligonucleotide consists of the sequence set forth 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 set forth in SEQ ID NO:1, wherein the antisense oligonucleotide consists of the sequence set forth 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 set forth in SEQ ID NO: 1, and the antisense oligonucleotide consists of any one of the sequences set forth in 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 set forth in SEQ ID NO: 1, and the antisense oligonucleotide is selected from the group consisting 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, 190, 200, 210, 212, 214, 216, 218, 220, 222, 224, 226, 228, 230, 232, 234, 235, 249, 150, 164, 165, 179, 180, 191, 219, 225, 232, 233, 234, 235, 249, 150, 164, 165, 179, 180, 192, 235, 246, 247, 248, 250, 252, 254, 256, 258, 260, 262, 264, 265, 270, 272, 274, 275, 276, 278, 279, 280, 281, 282, 283, 284, 285, 286, 287, 288, 289, 300, 301, 302, 303, 304, 305, 4, 195, 209, 210, 224, 225, 239, 240, 254, 255, 269, 270, 284, 285, 299, 300, 314, 329, 344, 518 to 638, 817, 818, 845, 855, 871, 882, 904, 958, 959, 960, 1024 to 1041, and 1200 to 1224. In some embodiments, the antisense oligonucleotide consists of 19 nucleotides complementary to the sequence set forth in SEQ ID NO: 1, and the antisense oligonucleotide consists of the sequence set forth 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 set forth in SEQ ID NO: 1, and the antisense oligonucleotide consists of the sequence set forth 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 set forth in SEQ ID NO: 1, and the antisense oligonucleotide consists of a sequence set forth 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 set forth in SEQ ID NO: 1, and the antisense oligonucleotide consists of any one of the sequences set forth in 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 set forth in SEQ ID NO: 1, and the antisense oligonucleotide consists of any one of the sequences set forth in 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 set forth in SEQ ID NO: 1, and the antisense oligonucleotide consists of any of SEQ ID NOs: 53-54, 68, 69, 83, 84, 98, 99, 113, 114, 128, 129, 143, 144, 158, 159, 173, 174, 188, 189, 203, 204, 205, 206, 207, 208, 209, 210, 211, 212, 213, 214, 215, 216, 217, 218, 219, 220, 221, 222, 223, 224, 225, 226, 227, 228, 229, 230, 231, 232, 233, 234, 235, 236, 237, 238, 239, 240, 241, 242, 243, 244, 245, 246, 247, 248, 249, 250, 251, 252, 253, 254, 255, 256, 257, 258, 259, 260, 261, 262, 263, 264, 265, 266, 267, 268, 269, 270, 271, 272, 273, 274, 275, 276, 2 18, 219, 233, 234, 248, 249, 263, 264, 278, 279, 293, 294, 308, 309, 323, 338, 353, 821, 822, 832 to 848, 858, 874, 887, 907, 919, 920, 923, 931, 946, 951, 1104 to 1115, and 1314 to 1332. In some embodiments, the antisense oligonucleotide consists of 25 nucleotides complementary to the sequence set forth in SEQ ID NO: 1, and the antisense oligonucleotide consists of any one of the sequences set forth in SEQ ID NOs: 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.

[0010] In some embodiments, the antisense oligonucleotide of the methods provided herein is complementary to the sequence set forth in any one of SEQ ID NOs: 700-703. In some embodiments, the antisense oligonucleotide has at least 80%, at least 90%, at least 95%, at least 98%, or 100% complementarity to the sequence set forth in any one of SEQ ID NOs: 700-703. In some embodiments, the antisense oligonucleotide has at least 80%, at least 90%, at least 95%, at least 98%, or 100% complementarity to the sequence set forth 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 to the sequence set forth in SEQ ID NO: 703.

[0011] In some embodiments, the antisense oligonucleotide of the methods provided herein is complementary to the sequence set forth in any one of SEQ ID NOs: 1339-1342, or a portion thereof. In some embodiments, the antisense oligonucleotide is complementary to the sequence set forth in SEQ ID NO: 1339, or a portion thereof. In some embodiments, the antisense oligonucleotide is complementary to the sequence set forth in SEQ ID NO: 1340, or a portion thereof. In some embodiments, the antisense oligonucleotide is complementary to the sequence set forth in SEQ ID NO: 1341, or a portion thereof. In some embodiments, the antisense oligonucleotide is complementary to the sequence set forth 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 set forth 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 set forth 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 set forth 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 set forth 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 set forth 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 set forth in SEQ ID NO: 1342.

[0012] In some embodiments, the cells express low levels of UNC13A before contacting the cells with the antisense oligonucleotide. In some embodiments, the method includes measuring the level of UNC13A protein in the cells. In some embodiments, the UNC13A protein level is measured by Western blot, imaging, ELISA, fluorescent reporter assay, luminescence assay, or immunohistochemistry.

[0013] In some embodiments, the secondary structure of UNC13A mRNA is altered by binding of the antisense oligonucleotide to the UNC13A transcript. In some embodiments, the antisense oligonucleotide sterically blocks the inclusion of an 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 to the sequence set forth in SEQ ID NO:1.

[0014] In some embodiments, the antisense oligonucleotides of the methods provided herein restore expression of UNC13A mRNA in a 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 a wild-type cell. In some embodiments, the antisense oligonucleotides increase expression of UNC13A mRNA in a cell 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 a sequence set forth in any one of SEQ ID NOs: 1059-1074, 1075-1089, 1090-1103, 1248-1270, 1271-1292, and 1293-1313.

[0015] In some embodiments, the antisense oligonucleotides of the methods provided herein reduce expression of UNC13A cryptic exon splice variant mRNA by at least 10%. In some embodiments, the antisense oligonucleotides reduce 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, or antisense oligonucleotides targeting TARDBP. In some embodiments, the antisense oligonucleotides reduce 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 any one of the sequences set forth in SEQ ID NOs: 1059-1074, 1075-1089, 1090-1103, 1248-1270, 1271-1292, and 1293-1313.

[0016] In some embodiments, the UNC13A cryptic exon splice variant comprises the sequence set forth in SEQ ID NO:7 or SEQ ID NO:9.

[0017] In some embodiments, after contacting the cells with the antisense oligonucleotide, the ratio of UNC13A transcripts that do not contain the cryptic exon splice variant to UNC13A transcripts that contain the cryptic exon splice variant is at least 10: 1. In some embodiments, the presence and / or amount of the cryptic exon splice variant is measured using qPCR.

[0018] In some embodiments, the cell is a neuron, hi some embodiments, the neuron is a motor neuron, a cortical neuron, an interneuron, or an excitatory neuron.

[0019] In some embodiments, contacting a cell with the antisense oligonucleotide of the methods provided herein increases 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 potentials, neuronal synaptic plasticity, and neuronal longevity.

[0020] In some embodiments, the antisense oligonucleotides of the methods provided herein restore expression of UNC13A protein in a 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 a wild-type cell. In some embodiments, the antisense oligonucleotides increase expression of UNC13A protein in a cell 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 a sequence set forth in any one of SEQ ID NOs: 1059-1074, 1075-1089, 1090-1103, 1248-1270, 1271-1292, and 1293-1313.

[0021] In some embodiments, the antisense oligonucleotide of the methods provided herein increases expression of UNC13A mRNA in a cell by at least 50%, decreases expression of UNC13A cryptic exon mRNA in a cell by at least 25%, and increases expression of UNC13A protein in a cell by at least 50%. In some embodiments, the antisense oligonucleotide consists of the sequence set forth 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 set forth in any one of SEQ ID NOs: 1059-1074, 1075-1089, and 1090-1103. In some embodiments, the antisense oligonucleotide consists of the sequence set forth in any one of SEQ ID NOs: 1248-1270, 1271-1292, and 1293-1313.

[0022] 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 repeats→3-5 CATC repeats), and rs62121687 (hg38 chr19:17,642,351 C→A), or any combination thereof. In some embodiments, the cells comprise a mutation associated with amyotrophic lateral sclerosis (ALS) or frontotemporal dementia (FTD).

[0023] In some embodiments, the antisense oligonucleotide is provided to the cell by transfection. In some embodiments, the cell expresses low levels of TDP-43 protein. In some embodiments, the cell is a human cell or a mouse cell.

[0024] In some embodiments, the antisense oligonucleotide of the method provided herein does not cause any changes in cell morphology.In some embodiments, the antisense oligonucleotide does not cause neurotoxicity.In some embodiments, the antisense oligonucleotide does not bind to genome sequences other than the sequence set forth in SEQ ID NO: 1.

[0025] Also provided herein is a method of treating amyotrophic lateral sclerosis (ALS) or frontotemporal dementia (FTD) in an individual, comprising administering to the individual an antisense oligonucleotide comprising 13 to 30 nucleotides, wherein the antisense oligonucleotide is complementary to the sequence set forth in SEQ ID NO:1, and 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 chr19:17,641,880 A→C), rs12973192 (hg38 chr19:17,642,430 C→G), rs56041637 (hg38 chr19:17,642,033-17,642,056 0-2 CATC repeats→3-5 CATC repeats), and rs62121687 (hg38 chr19:17,642,351 C→A), or any combination thereof. In some embodiments, the mutation associated with ALS or FTD is a mutation in TDP43.

[0026] In some embodiments, the antisense oligonucleotide has a sequence that is complementary to a sequence in the 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. 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.

[0027] Also provided herein is an antisense oligonucleotide that regulates UNC13A splicing, wherein the antisense oligonucleotide has a sequence complementary to a 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. Also provided herein is an antisense oligonucleotide that regulates UNC13A splicing, comprising 13 to 30 nucleotides, wherein the antisense oligonucleotide comprises a sequence set forth in any one of SEQ ID NOs: 20 to 355, 400 to 638, 810 to 959, or 961 to 1332.

[0028] In some embodiments, the antisense oligonucleotide is complementary to the sequence set forth in SEQ ID NO: 1. In some embodiments, the antisense oligonucleotide is complementary to the sequence set forth 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 set forth 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 set forth 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 set forth in SEQ ID NO: 703.

[0029] In some embodiments, the antisense oligonucleotide is complementary to the sequence set forth in any one of SEQ ID NOs: 1339-1342, or a portion thereof. In some embodiments, the antisense oligonucleotide is complementary to the sequence set forth in SEQ ID NO: 1339, or a portion thereof. In some embodiments, the antisense oligonucleotide is complementary to the sequence set forth in SEQ ID NO: 1340, or a portion thereof. In some embodiments, the antisense oligonucleotide is complementary to the sequence set forth in SEQ ID NO: 1341, or a portion thereof. In some embodiments, the antisense oligonucleotide is complementary to the sequence set forth 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 set forth 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 set forth 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 set forth 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 set forth 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 set forth 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 set forth in SEQ ID NO:1342.

[0030] Also provided herein are antisense oligonucleotides comprising a sequence set forth in any one of SEQ ID NOs: 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 set forth in SEQ ID NO: 1.

[0031] Also provided herein is an antisense oligonucleotide consisting of a sequence set forth in any one of SEQ ID NOs: 20 to 355, 400 to 638, 810 to 959, or 961 to 1332. Also provided herein is an antisense oligonucleotide that is complementary to a sequence set forth in any one of SEQ ID NOs: 1339 to 1342 or a portion thereof.

[0032] In some embodiments, the antisense oligonucleotide consists of 13 nucleotides complementary to the sequence set forth in SEQ ID NO:1, wherein the antisense oligonucleotide consists of the sequence set forth 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 set forth in SEQ ID NO:1, wherein the antisense oligonucleotide consists of the sequence set forth 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 set forth in SEQ ID NO:1, wherein the antisense oligonucleotide consists of the sequence set forth 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 set forth in SEQ ID NO: 1, and the antisense oligonucleotide consists of any one of the sequences set forth in 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 set forth in SEQ ID NO: 1, and the antisense oligonucleotide is selected from the group consisting 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, 190, 200, 210, 212, 214, 216, 218, 220, 222, 224, 226, 228, 230, 232, 234, 235, 249, 150, 164, 165, 179, 180, 191, 219, 225, 232, 233, 234, 235, 249, 150, 164, 165, 179, 180, 192, 235, 246, 247, 248, 250, 252, 254, 256, 258, 260, 262, 264, 265, 270, 272, 274, 275, 276, 278, 279, 280, 281, 282, 283, 284, 285, 286, 287, 288, 289, 300, 301, 302, 303, 304, 305, 4, 195, 209, 210, 224, 225, 239, 240, 254, 255, 269, 270, 284, 285, 299, 300, 314, 329, 344, 518 to 638, 817, 818, 845, 855, 871, 882, 904, 958, 959, 960, 1024 to 1041, and 1200 to 1224. In some embodiments, the antisense oligonucleotide consists of 19 nucleotides complementary to the sequence set forth in SEQ ID NO: 1, and the antisense oligonucleotide consists of the sequence set forth 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 set forth in SEQ ID NO: 1, and the antisense oligonucleotide consists of the sequence set forth 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 set forth in SEQ ID NO: 1, and the antisense oligonucleotide consists of a sequence set forth 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 set forth in SEQ ID NO: 1, and the antisense oligonucleotide consists of any one of the sequences set forth in 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 set forth in SEQ ID NO: 1, and the antisense oligonucleotide consists of any one of the sequences set forth in 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 set forth in SEQ ID NO: 1, and the antisense oligonucleotide consists of any of SEQ ID NOs: 53, 54, 68, 69, 83, 84, 98, 99, 113, 114, 128, 129, 143, 144, 158, 159, 173, 174, 188, 189, 203, 204, 205, 206, 207, 208, 209, 210, 211, 212, 213, 214, 215, 216, 217, 218, 219, 220, 221, 222, 223, 224, 225, 226, 227, 228, 229, 230, 231, 232, 233, 234, 235, 236, 237, 238, 239, 240, 241, 242, 243, 244, 245, 246, 247, 248, 249, 250, 251, 252, 253, 254, 255, 256, 257, 258, 259, 260, 261, 262, 263, 264, 265, 266, 267, 268, 269, 270, 271, 272, 273, 274, 275, 276, 18, 219, 233, 234, 248, 249, 263, 264, 278, 279, 293, 294, 308, 309, 323, 338, 353, 821, 822, 832 to 848, 858, 874, 887, 907, 919, 920 to 923, 931, 946 to 951, 1104 to 1115, and 1314 to 1332. In some embodiments, the antisense oligonucleotide consists of 25 nucleotides complementary to the sequence set forth in SEQ ID NO: 1, and the antisense oligonucleotide consists of any one of the sequences set forth in SEQ ID NOs: 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.

[0033] In some embodiments, the antisense oligonucleotide is complementary to the sequence set forth in SEQ ID NO: 1339, or a portion thereof. In some embodiments, the antisense oligonucleotide is complementary to the sequence set forth in SEQ ID NO: 1340, or a portion thereof. In some embodiments, the antisense oligonucleotide is complementary to the sequence set forth in SEQ ID NO: 1341, or a portion thereof. In some embodiments, the antisense oligonucleotide is complementary to the sequence set forth 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 set forth in any one of SEQ ID NOs: 1339-1342.

[0034] 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 set forth 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 set forth 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 set forth 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 set forth 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 set forth in SEQ ID NO: 1342. In some embodiments, the antisense oligonucleotide consists of the sequence set forth in any one of SEQ ID NOs: 1059-1074, 1075-1089, 1090-1103, 1248-1270, 1271-1292, and 1293-1313.

[0035] 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 / or 3'-end of the antisense oligonucleotide comprises a modification, and 2, 3, 4, or 5 nucleotides are modified at each of the 5'-end and / or 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 oligonucleotides, 2'O-methoxyethyl antisense oligonucleotides, phosphorothioate antisense oligonucleotides, and locked nucleic acid (LNA) antisense oligonucleotides. In some embodiments, the modification is a 2'Ome antisense oligonucleotide.

[0036] Also provided herein is a pharmaceutical composition comprising any one of the antisense oligonucleotides of the embodiments disclosed herein. In some embodiments, the pharmaceutical composition comprises an excipient and / or a buffer.

[0037] Also provided herein are kits comprising the antisense oligonucleotides of some embodiments disclosed herein or the pharmaceutical compositions of some embodiments disclosed herein.

[0038] All publications, including patent documents, scientific articles, and databases, referred to in this application are incorporated by reference in their entirety for all purposes to the same extent as if each individual publication were individually incorporated by reference. To the extent that a definition set forth herein contradicts or otherwise conflicts with a definition set forth in a patent, application, published application, or other publication incorporated herein by reference, the definition set forth herein takes precedence over the definition incorporated herein by reference.

[0039] The section headings used herein are for organizational purposes only and are not to be construed as limiting the subject matter described. [Brief explanation of the drawings]

[0040] 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.

[0041] [Figure 1A] Figure 1 shows the percent UNC13A mRNA upon treatment with ASO compared to treatment with a control ASO, which has no effect on UNC13A mRNA in motor neurons derived from TDP-43 knockdown iPSCs. UNC13A mRNA was detected using qPCR. The position of the ASO target is plotted from 5' to 3'. [Figure 1B] Same as above. [Figure 2A] Figure 1 shows the percent of UNC13A mRNA containing the cryptic exon (CE) upon treatment with ASO compared to treatment with a control ASO, which has no effect on UNC13A CE mRNA in motor neurons derived from TDP-43 knockdown iPSCs. qPCR was used to detect UNC13A CE mRNA. The position of the ASO target is plotted from 5' to 3'. [Figure 2B] Same as above. [Figure 3A] Figure 1 shows the percentage of UNC13A mRNA in motor neurons derived from TDP-43 knockdown iPSCs, the percentage of UNC13A mRNA containing the cryptic exon (CE), and the expression of the control transcript STMN2, which is not targeted by ASO, in motor neurons derived from TDP-43 knockdown iPSCs. The position of the ASO target is plotted from 5' to 3'. [Figure 3B] Same as above. [Figure 4]The percentage of UNC13A cryptic exon transcript and the percentage of UNC13A full-length transcript for each ASO tested in motor neurons derived from TDP-43 knockdown iPSCs are shown. ASOs that reduced UNC13A CE and increased UNC13A full-length transcript are shown as squares. [Figure 5A] 5B is a map of the UNC13A cryptic exons showing the location of active oligos shown in dark grey and inactive oligos shown in light grey. FIG. 5B shows the four identified regulatory regions. [Figure 5B] Same as above. [Figure 6A] Effect of ASOs on UNC13A expression in motor neurons derived from TDP43-knockdown iPSCs. NT, non-targeting ASO; ASO434, inactive ASO; +NT, non-targeting shRNA; +shTDP43, shRNA against TARDBP. [Figure 6B] Same as above. [Figure 6C] Figure 1 shows the effect of ASO on TARDBP expression in iPSC-derived motor neurons treated with shRNA to TDP-43 compared with control cells treated with an inactive ASO. [Figure 6D] The effect of ASOs on STMN2 expression in iPSC-derived motor neurons treated with shRNA to TDP-43 compared with control cells treated with an inactive ASO is shown. NT, non-targeting ASO; ASO434, inactive ASO; +NT, non-targeting shRNA; +shTDP43, shRNA to TARDBP. [Figure 7A] 1 shows the effect of ASO422 on percent UNC13A mRNA expression in iPSC-derived motor neurons with TDP-43 knockdown in outgrowth cultures compared to ASOs that do not target UNC13A or control ASOs. [Figure 7B] 1 shows the effect of ASO422 on UNC13A cryptic exons (CEs) in iPSC-derived motor neurons with TDP-43 knockdown in outgrowth cultures compared to ASOs that do not target UNC13A or control ASOs. [Figure 7C]1 shows the effect of ASO422 on percent TARDBP expression in iPSC-derived motor neurons with TDP-43 knockdown in outgrowth cultures compared to ASOs not targeting UNC13A or control ASOs. [Figure 8A] Western blot of UNC13 protein levels in iPSC-derived motor neurons with TDP-43 knockdown treated with ASO or a control ASO (432) that does not target UNC13A. [Figure 8B] Quantitation of UNC13A protein levels as determined by Western blot is shown. [Figure 9A] Shown is the percentage of UNC13A mRNA expression after treatment with various doses of ASO (0.024-3 μM) compared to control ASO, which has no effect on UNC13A mRNA expression. [Figure 9B] Same as above. [Figure 9C] The percentage of UNC13A mRNA containing the cryptic exon (CE) after treatment with various doses of ASO (0.024–3 μM) compared with the control ASO, which has no effect on UNC13A mRNA expression, is shown. [Figure 9D] Same as above. [Figure 9E] Figure 1 shows TARDBP and STMN2 expression levels after treatment with various doses of ASO. After ASO delivery, cells were transduced with shRNA against TARDBP or a control. qPCR was used to detect UNC13A, UNC13A CE, STMN2, and TARDBP mRNA expression levels. ASO432, inactive ASO; +NT, non-targeting shRNA; +shTDP43, shRNA against TARDBP. [Figure 9F] Same as above. [Figure 10A] Shown is the percent of UNC13A protein expression for individual ASO treatments at various concentrations (0.024-3 µM) in iPSC-derived motor neurons with TDP-43 knockdown. [Figure 10B] Percent UNC13A protein expression for all ASO treatment conditions is shown. [Figure 10C] The rank order of HiBiT signal (UNC13A protein expression) for 3 μM ASO-treated neurons is shown. [Figure 10D] The percent of UNC13A mRNA and UNC13A protein expression for each control and UNC13A splice-correcting ASO tested is shown. [Figure 11A] Shown is the percentage of UNC13A mRNA expression after treatment with ASO (0.024 μM or 3 μM) compared to control ASO, which does not affect UNC13A mRNA expression in iPSC-derived motor neurons with TDP-43 knockdown. [Figure 11B] Shown is the percentage of UNC13A protein expression after treatment with ASO (0.024 μM or 3 μM) compared to control ASO, which does not affect UNC13A mRNA expression. [Figure 11C] Percentages of UNC13A mRNA and UNC13A protein expression are shown for each control and UNC13A splice-correcting ASO tested. After ASO delivery, cells were transduced with shRNA against TARDBP or control. qPCR was used to detect UNC13A, UNC13A CE, STMN2, and TARDBP mRNA expression levels. ASO432, inactive ASO; +NT, non-targeting shRNA; +shTDP43, shRNA against TARDBP. [Figure 12A] Figure 1 shows the percent of UNC13A and UNC13A CE mRNA expression after treatment with ASO (0.024 μM or 3 μM) compared to a control ASO, which 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. [Figure 12B] Same as above. [Figure 13A]Figure 1 shows the percent of UNC13A and UNC13A CE mRNA expression after treatment with ASO (0.024 μM or 3 μM) compared to a control ASO, which 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. [Figure 13B] Same as above. [Figure 14A] Figure 1 shows the percent of UNC13A and UNC13A CE mRNA expression after treatment with ASO (0.024 μM or 3 μM) compared to a control ASO, which 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. [Figure 14B] Same as above. [Figure 15A] Figure 1 shows the percent of UNC13A and UNC13A CE mRNA expression after treatment with ASO (0.024 μM or 3 μM) compared to a control ASO, which 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. [Figure 15B] Same as above. [Figure 16A] Cumulative UNC13A and UNC13A CE mRNA expression data from four iPSC lines (GM, 005, 4048, and 4157) are shown. Percentages of UNC13A and UNC13A CE mRNA expression after treatment with UNC13A splice-blocking ASO (0.024 μM or 3 μM) are plotted. After ASO delivery, cells were transduced with shRNA against TARDBP or a control. [Figure 16B] Same as above. [Figure 17A]Figures 17A and 17B show the percentages of UNC13A mRNA expression (Figures 17C and 17D), UNC13A CE mRNA expression (Figures 17E and 17G), and STMN2 mRNA expression (Figures 17F and 17H) after long-term culture and multiple doses of UNC13A splice-correcting ASO or control non-targeting ASO (432) in GM and 005 iPSC-derived motor neurons. GM and 005 cells were treated with a gapmer ASO against TARDBP or a non-targeting control gapmer before ASO administration to correct UNC13A splice. [Figure 17B] Same as above. [Figure 17C] Same as above. [Figure 17D] Same as above. [Figure 17E] Same as above. [Figure 17F] Same as above. [Figure 17G] Same as above. [Figure 17H] Same as above. [Figure 18A] Percent nuclear area of ​​neurons treated with UNC13A splice-blocking ASO is shown. [Figure 18B] Images of neurons after control or ASO treatment are shown. [Figure 19A] Figures 19A (13mer), 19C (15mer), 19E (17mer), 19G (18mer), 19I (19mer), 19K (20mer), 19M (21mer), 19O (22mer), and 19Q (24mer) show the percentage of UNC13A and UNC13A CE mRNA expression after treatment with ASOs of different lengths (3 μM) targeting region 2 of UNC13A CE. Figure 19B (13mer), Figure 19D (15mer), Figure 19F (17mer), Figure 19H (18mer), Figure 19J (19mer), Figure 19L (20mer), Figure 19N (21mer), Figure 19P (22mer), and Figure 19R (24mer) show the percent UNC13A CE mRNA expression. iPSC-derived motor neurons were transduced with shRNA versus TARDBP or control after ASO delivery. [Figure 19B] Same as above. [Figure 19C] Same as above. [Figure 19D] Same as above. [Figure 19E] Same as above. [Figure 19F] Same as above. [Figure 19G] Same as above. [Figure 19H] Same as above. [Figure 19I] Same as above. [Figure 19J] Same as above. [Figure 19K] Same as above. [Figure 19L] Same as above. [Figure 19M] Same as above. [Figure 19N] Same as above. [Figure 19O] Same as above. [Figure 19P] Same as above. [Figure 19Q] Same as above. [Figure 19R] Same as above. [Figure 20A] Figures 20A (13mer), 20C (15mer), 20E (17mer), 20G (18mer), 20I (19mer), 20K (20mer), 20M (21mer), 20O (22mer), and 20Q (24mer) show the percentage of UNC13A and UNC13A CE mRNA expression after treatment with ASOs of different lengths (3 μM) targeting region 4 of UNC13A CE. Figure 20B (13mer), Figure 20D (15mer), Figure 20F (17mer), Figure 20H (18mer), Figure 20J (19mer), Figure 20L (20mer), Figure 20N (21mer), Figure 20P (22mer), and Figure 20R (24mer) show the percent UNC13A CE mRNA expression. iPSC-derived motor neurons were transduced with shRNA versus control after TARDBP or ASO delivery. [Figure 20B] Same as above. [Figure 20C] Same as above. [Figure 20D] Same as above. [Figure 20E] Same as above. [Figure 20F] Same as above. [Figure 20G] Same as above. [Figure 20H] Same as above. [Figure 20I] Same as above. [Figure 20J] Same as above. [Figure 20K] Same as above. [Figure 20L] Same as above. [Figure 20M] Same as above. [Figure 20N] Same as above. [Figure 20O] Same as above. [Figure 20P] Same as above. [Figure 20Q] Same as above. [Figure 20R] Same as above. [Figure 21A] Shown is the average percentage of UNC13A mRNA expression from all cells treated with the indicated ASO lengths contained within the sequence on the left. Darker shading indicates higher UNC13A mRNA expression. [Figure 21B] The average difference in percent UNC13A mRNA expression between cells treated with the parental 24mer ASO shown on the left and all cells treated with the indicated daughter ASOs of the length contained within the parental sequence is shown. Lighter shading indicates increased UNC13A mRNA expression for the daughter ASO of that length compared to the parental 24mer, and darker shading indicates decreased UNC13A mRNA expression for the daughter ASO of that length compared to the parental 24mer. After ASO delivery, cells were transduced with shRNA against TARDBP or a control. [Figure 22A] Figure 22 shows the effect of ASOs ranging from 13 nucleotides (13mer) to 24 nucleotides (24mer) tiling region 2 (Figure 22A) and region 4 (Figure 22B) on UNC13A mRNA expression in cells transduced with shRNA against TARDBP after ASO delivery. The top of the panel plots the average percent UNC13A expression for all ASOs overlapping each indicated nucleotide, grouped by ASO length. [Figure 22B] Same as above. DETAILED DESCRIPTION OF THE INVENTION

[0042] TDP-43 depletion is associated with several neuropathologies, including ALS and FTD. TDP-43 depletion leads to incorrect splicing of UNC13A mRNA and the inclusion of a cryptic exon between exons 20 and 21. The inclusion of the UNC13A cryptic exon results in a decrease in UNC13A protein and is associated with such neuropathologies. Provided herein are antisense oligonucleotides targeting UNC13A mRNA that reduce the 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 located within the cryptic exon itself. The antisense oligonucleotides can restore UNC13A expression in neurons and increase the ratio of correctly spliced ​​to incorrectly spliced ​​UNC13A transcripts.

[0043] definition Before describing this disclosure in more detail, it may be helpful to an understanding thereof to provide definitions of certain terms used herein. Additional definitions are set forth throughout this disclosure.

[0044] In this description, unless otherwise indicated, any concentration range, percentage range, ratio range, or integer range should be understood to include any integer value within the recited range, and, where appropriate, fractions thereof (such as integer tenths and hundredths) or subranges. The description of a range includes the description of each endpoint of the range.

[0045] As used herein, the term "about" means ±20% of the indicated range, value, or structure, unless otherwise indicated.

[0046] As used herein, the terms "a" and "an" should be understood to refer to "one or more" of the listed components. The use of the alternative (e.g., "or") should be understood to mean either one, both, or any combination thereof of the alternatives.

[0047] As used herein, the terms "include," "having," and "comprise" are used interchangeably and such terms and variations are intended to be interpreted as open-ended.

[0048] "Optional" or "optionally" means that the described element, component, event, or circumstance may or may not occur, and the description includes instances in which the element, component, event, or circumstance occurs as well as instances in which it does not occur.

[0049] As used herein, "nucleic acid" or "nucleic acid molecule" or "polynucleotide" refers to any of deoxyribonucleic acid (DNA), ribonucleic acid (RNA), oligonucleotides, such as molecules produced by polymerase chain reaction (PCR) or by in vitro translation, and molecules produced by either ligation, cleavage, endonuclease action, exonuclease action, or mechanical action (e.g., shearing). Nucleic acids can be composed of multiple monomers that are naturally occurring nucleotides (such as deoxyribonucleotides and ribonucleotides), analogs of naturally occurring nucleotides (e.g., α-enantiomeric forms of naturally occurring nucleotides), or combinations of both. Modified nucleotides can have modifications or substitutions of sugar moieties, or pyrimidine or purine base moieties (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, phosphorodithioates, phosphoroselenates, phosphorodiselenates, phosphorothioates, phosphoroanylioates, phosphoranilidates, phosphoramidates, etc. Nucleic acid molecules can be either single-stranded or double-stranded.

[0050] As used herein, "protein" or "polypeptide" refers to a compound consisting of amino acid residues covalently linked by peptide bonds. The term "protein" may be synonymous with the term "polypeptide" or may additionally refer to a complex of two or more polypeptides. In certain embodiments, a polypeptide may be a fragment. As used herein, a "fragment" refers to 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 reference 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.

[0051] The term "isolated" means that a material, complex, compound, or molecule is removed from its original environment (e.g., the natural environment if it occurs in nature). 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 of the natural system is isolated. Such a nucleic acid may be part of a vector, and / or such a nucleic acid or polypeptide may be part of a composition (e.g., a cell lysate), but such a vector or composition is still isolated in that it is not part of the natural environment for the nucleic acid or polypeptide. The term "gene" means a segment of DNA involved in producing a polypeptide chain, including regions preceding and following the coding region "leader and trailer," as well as intervening sequences (introns), if present, between individual coding segments (exons).

[0052] 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 refers to a cell or microorganism that has been modified by human or machine introduction of an exogenous or heterologous nucleic acid molecule, or that has been modified by human or machine intervention such that expression of an endogenous nucleic acid molecule or gene is controlled, deregulated, or constitutive. Human-generated genetic modifications can include, for example, modifications that introduce additions, deletions, substitutions, or other functional disruptions or additions of nucleic acid molecules (which may include expression control elements such as promoters) encoding one or more proteins or enzymes, or other nucleic acid molecules of a cell's genetic material or encoded products. Exemplary human- or machine-introduced modifications include modifications in the coding region of a heterologous or homologous polypeptide or functional fragment thereof from a reference or parent molecule.

[0053] A "wild-type" gene or gene product is that which is most frequently observed in a population and is thus arbitrarily designed the "normal" or "reference" or "wild-type" form of the gene.

[0054] As used herein, a "mutation" refers to a change in the sequence of a nucleic acid molecule or polypeptide molecule compared to a reference or wild-type nucleic acid molecule or polypeptide molecule, respectively. Mutations can result in several different types of changes in the sequence, including nucleotide or amino acid substitutions, insertions, or deletions.

[0055] A "conservative substitution" refers to an amino acid substitution that does not significantly affect or alter the binding characteristics of a particular protein. Generally, a conservative substitution is one in which the substituted amino acid residue 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 by similar function, chemical structure, or composition (e.g., acidic, basic, aliphatic, aromatic, or sulfur-containing). For example, aliphatic groups can include, for substitution purposes, Gly, Ala, Val, Leu, and Ile. Other conservative substitution groups include sulfur-containing groups: Met and cysteine ​​(Cys or C); acidic groups: 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, WH Freeman and Company.

[0056] As used herein, the term "expression" refers to the process by which a polypeptide is produced based on a coding sequence of a nucleic acid molecule such as a gene. The process may include transcription, post-transcriptional regulation, post-transcriptional modification, translation, post-translational regulation, post-translational modification, or any combination thereof.

[0057] As used herein, "sequence identity" refers to the percentage of nucleotides (amino acid residues) in a sequence that are identical to those in another reference polynucleotide (polypeptide) sequence, after aligning the sequences and introducing gaps, if necessary, to achieve the maximum percent sequence identity, without considering conservative substitutions as part of the sequence identity.Percentage sequence identity values ​​can be generated using the NCBI BLAST 2.0 software, as defined by Altschul et al. (1997) "Gapped BLAST and PSI-BLAST: a new generation of protein database search programs," Nucleic Acids Res. 25:3389-3402, with parameters set to default values.

[0058] As used herein, "UNC13A" refers to a presynaptic protein found in central and neuromuscular synapses that regulates the release of neurotransmitters, peptides, and hormones. The UNC13A reference or wild-type mRNA transcript contains 44 exons encoding a 1,703 amino acid protein. In embodiments, the NCBI reference sequence: NP_001073890.2 (SEQ ID NO: 11) is an example of a wild-type or reference UNC13A protein. In embodiments, the NCBI reference sequence NM_001080421.3 (SEQ ID NO: 1) is an example of a wild-type or reference UNC13A mRNA transcript. In embodiments, UNC13A includes all forms of UNC13A, including wild-type, splice isoforms, variants, mutants, native conformations, misfolding, and post-translational modifications. In embodiments, UNC13A excludes UNC13A cryptic exon splice variants.

[0059] As used herein, the term "preprocessed mRNA" or "pre-mRNA" or "precursor mRNA" refers to a primary transcript synthesized from transcription of a DNA template that has not undergone processing to become a mature mRNA, e.g., splicing, addition of a 5' cap, and addition of a 3' polyA tail. Mature mRNA can be translated into protein by ribosomes.

[0060] As used herein, the term "cryptic exon" or "pseudo-exon" refers to an exon that is not present or detectably used in the wild-type pre-mRNA but is selected for in a variant isoform; cryptic exons can arise as a result of mutations that create new splice sites or remove existing binding sites for splicing repressors. Cryptic exons can also emerge from transposable elements (e.g., Alu elements).

[0061] As used herein, "UNC13A cryptic exon splice variant" refers to an mRNA, or a protein encoded by such an mRNA, that includes a cryptic exon between exons 20 and 21. The cryptic exon is obtained from introns 20-21 of the UNC13A gene. In embodiments, the cryptic exon has the nucleotide sequence of SEQ ID NO: 5 or SEQ ID NO: 6. In embodiments, the UNC13A cryptic exon splice variant may have the nucleotide sequence of SEQ ID NO: 7, which encodes the protein sequence of SEQ ID NO: 8, or the nucleotide sequence of SEQ ID NO: 9, which encodes the protein sequence of SEQ ID NO: 10.

[0062] As used herein, "transactivation response element DNA-binding protein 43" or "TAR-DNA-binding protein-43" or "TDP-43" refers to the typically 414 amino acid residue protein encoded by TARDBP, as well as the mRNA transcript of TARDBP. In certain instances, the mRNA encoded by the TARDBP gene is referred to herein as TDP-43. In embodiments, the wild-type TDP-43 amino acid sequence is provided by Uniprot Accession No. Q13148 (SEQ ID NO: 378). In embodiments, TDP-43 includes all forms of TDP-43, including wild-type, splice isoforms, variants, mutants, native conformations, misfolded, and post-translationally modified (e.g., ubiquitinated, phosphorylated, acetylated, sumoylated, or truncated to a C-terminal fragment) proteins.

[0063] 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. Cytoplasmic inclusions of hyperphosphorylated, ubiquitinated, and truncated forms of TDP-43 are pathological hallmarks of diseases, 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 neuropathy (FOSMN), hippocampal sclerosis (HS), limbic-dominant age-related TDP-43 encephalopathy (LATE), brain age-related TDP-43 with sclerosis (CARTS), Guam Parkinson-Dementia Complex (G-PDC), Guan ALS (G-ALS), multisystem proteinopathy (MSP), Perry disease, Alzheimer's disease (AD), and chronic traumatic encephalopathy (CTE).

[0064] The terms "complementary" and "complementarity" refer to polynucleotides (i.e., a sequence of nucleotides) related by the base-pairing rules. For example, the sequence "AGT" is complementary to the sequence "TCA." Complementarity can be "partial," where only some of the nucleic acids' bases match according to the base-pairing rules, or there can be "complete" or "total" complementarity between nucleic acids. The degree of complementarity between nucleic acid strands has a significant effect on the efficiency and strength of hybridization between nucleic acid strands. While perfect complementarity is often desired, some embodiments include one or more, but preferably 6, 5, 4, 3, 2, or 1, mismatches with respect to the target nucleic acid (e.g., RNA). Variation at any position within the oligomer can be included. In certain embodiments, sequence variation near the termini of the oligomer is generally preferred over internal variation, and, if present, is typically within about 6, 5, 4, 3, 2, or 1 nucleotide of the 5' and / or 3' end.

[0065] 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) by Watson-Crick base pairing to form a nucleic acid:oligomer heteroduplex within the target sequence. Antisense oligonucleotides may contain unmodified nucleotides, or may contain modified, non-natural, or analogous nucleotides, such as morpholinos, phosphorothioates, peptide nucleic acids, LNAs, 2'-O-Me RNA, 2'F-RNA, 2'-O-MOE-RNA, 2'F-ANA, or any combination thereof.

[0066] Such antisense oligomers can be designed to block or inhibit translation of mRNA, inhibit natural pre-mRNA splice processing, or induce degradation of the target mRNA, and may be said to be "directed" or "targeted" to the target sequence to which they hybridize. In embodiments, the target sequence is the region surrounding or including the AUG start codon of the mRNA, the 3' or 5' splice site of the pre-processed mRNA, or the branch point. The target sequence can be within an exon or an intron, or a combination thereof. A splice site target sequence can include an mRNA sequence having its 5' end 1 to about 25 base pairs downstream of the normal splice acceptor junction in the pre-processed mRNA. Exemplary splice site target sequences are any region of the pre-processed mRNA that includes the splice site, is entirely contained within the exon coding sequence, or spans the splice acceptor or donor site. More generally, in this disclosure, oligomers are said to be "targeted" to a biologically relevant target, such as the human UNC13A gene pre-mRNA encoding the UNC13A protein, which is targeted to the target nucleic acid in the manner described above. Exemplary targeting sequences include those listed in Tables 2-5.

[0067] The term "oligonucleotide analog" refers to an oligonucleotide having (i) a modified backbone structure, e.g., a backbone other than the standard phosphodiester linkage found in natural oligo- and polynucleotides, and (ii) an optionally modified sugar moiety, e.g., a morpholino moiety rather than a ribose or deoxyribose moiety. Oligonucleotide analogs bear bases that can hydrogen bond to standard polynucleotide bases through Watson-Crick base pairing, and the analog backbone presents the bases in a manner that allows such hydrogen bonding in a sequence-specific manner between the oligonucleotide analog molecule and the bases in a standard polynucleotide (e.g., single-stranded RNA or single-stranded DNA). Exemplary analogs are those that have a substantially uncharged phosphorus-containing backbone.

[0068] As used herein, a "modified nucleotide" is a nucleotide other than a ribonucleotide (2'-hydroxyl nucleotide). 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 mimics, abasic nucleotides (represented herein as x or ab), 2'-modified nucleotides, 3' to 3' linked (inverted) nucleotides (represented herein as invdn, invn, invx), unnatural base-containing nucleotides, bridged nucleotides, peptide nucleic acids, 2',3' seco nucleotide mimics (unlocked nucleobase analogs, represented herein as nuna or nuna), locked nucleotides (represented herein as nlna or nlna), 3'-o-methoxy (2' internucleotide linked) nucleotides (represented herein as 3'-omen), 2'-f-arabinonucleotides (represented herein as nfana or nfana), morpholino nucleotides, vinylphosphonate deoxyribonucleotides (represented herein as vpdn), and vinylphosphonate 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 five-membered sugar ring) include, but are not limited to, 2'omethyl nucleotides (represented herein as a lowercase "n" in nucleotide sequences), 2'-deoxy-2'-fluoro nucleotides (represented herein as nf, also referred to as 2'-fluoro nucleotides), 2'-deoxy nucleotides (represented herein as dn), 2'-methoxyethyl (2'-o-2-methoxyethyl) nucleotides (represented herein as nm or 2'-moe), 2'-amino nucleotides, and 2'-alkyl nucleotides. Not all positions in a given compound need be uniformly modified.Conversely, two or more modifications may be incorporated into a single ASO or into a single nucleotide. ASOs may be synthesized and / or modified by methods known in the art. A modification at one nucleotide is independent of a modification at another nucleotide.

[0069] An oligonucleotide "subunit" refers to one nucleotide (or nucleotide analog) unit that comprises a purine or pyrimidine base pair moiety. The term can refer to a nucleotide unit with or without an associated intersubunit linkage, although when referring to a "charged subunit," the charge is typically present in the intersubunit linkage (e.g., a phosphate or phosphorothioate linkage or a cation linkage).

[0070] 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 pyridin-4-one, pyridin-2-one, phenyl, pseudouracil, 2,4,6-trimethyl-1,1-thoxybenzene, 3-methyluracil, dihydrouridine, naphthyl, aminophenyl, 5-alkylcytidine (e.g., 5-methylcytidine), 5-alkyluridine (e.g., ribothymidine), 5-halouridine (e.g., 5-bromidine), or 6-azapyrimidine or 6-alkylpyrimidine (e.g., 6-methyluridine), propyne, kesosin, 2-thiouridine, 4-thiouridine, wybutoxosin, wybutoxosin, 4-acetylcytidine, 5-(carboxyhydroxymethyl)uridine, 5'-carboxymethylaminomethyl-2-thiouridine, and the like. Included are bases such as uridine, 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-methylcarbonifeneethyluridine, 5-methyloxyuridine, 5-methyl-2-thiouridine, 2-methylthio-N6-isopentenyladenosine, β-D-mannosylqueosine, uridine-5-oxyacetic acid, 2-thiocytidine, and threonine derivatives (Burgin et al., 1996, Biochemistry, 35:14090; Uhlman & Peyman, supra). In this embodiment, "modified base" refers to a nucleotide base other than adenine (A), guanine (G), cytosine (C), thymine (T), and uracil (U), as defined above, and such bases can be used at any position within an antisense molecule. Those skilled in the art will understand that Ts and Us are interchangeable, depending on the use of the oligomer. For example, in other antisense chemistries, such as 2'-O-methyl antisense oligonucleotides, which are RNA-like, a T base may be represented as U.

[0071] The term "target sequence" refers to a sequence in an oligomer or oligomer analog that is complementary (and thus essentially complementary) to a "target sequence" in an RNA genome. The entire sequence of an antisense oligomer may be complementary to a target sequence. For example, in an oligomer having 20-30 bases, approximately 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 may be a target sequence complementary to the target region. Typically, a target sequence is formed from consecutive bases in an oligomer, but alternatively, it may be formed from non-contiguous sequences that, when placed together, e.g., from opposite ends of the oligomer, constitute a sequence spanning the target sequence.

[0072] A "target sequence" can have "near" or "substantial" complementarity to a target sequence and still function for purposes of this disclosure, i.e., still be "complementary." Preferably, the oligomeric analog compounds used in this disclosure have at most one mismatch with the target sequence in 10 nucleotides, 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 exemplary target sequences being specified herein.

[0073] 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 may include hydrogen) and n is 1 to 7, preferably 1 to 4.

[0074] The term "naturally occurring amino acids" refers to amino acids present in naturally occurring proteins, such as the 20 (L)-amino acids utilized during protein biosynthesis, as well as other amino acids such as 4-hydroxyproline, hydroxylysine, desmosine, isodesmosine, homocysteine, citrulline, and ornithine. The term "unnatural amino acids" refers to amino acids that do not occur in naturally occurring proteins, and examples include β-alanine (β-Ala), 6-aminohexanoic acid (Ahx), and 6-aminopentanoic acid. Additional examples of "unnatural amino acids" include, but are not limited to, the (D)-amino acids known to those skilled in the art, such as norleucine, norvaline, p-fluorophenylalanine, and ethionine.

[0075] The term "target sequence" refers to the portion of a target RNA to which an oligonucleotide or antisense agent is directed, i.e., the sequence to which the oligonucleotide hybridizes by Watson-Crick base pairing of complementary sequences. In embodiments, the target sequence may be a contiguous region of a pre-mRNA that includes both intronic and exon target sequences. In embodiments, the target sequence will be exclusive of either intronic or exon sequences.

[0076] A target sequence and a targeting sequence are described as being "complementary" to each other when hybridization occurs in an antiparallel configuration.A target sequence may have "close" or "substantial" complementarity to a target sequence and still function for the purposes of the present disclosure, i.e., it can still be functionally "complementary".In certain embodiments, an oligonucleotide may have at most one mismatch with a target sequence in 10 nucleotides, and preferably at most one mismatch in 20 nucleotides.Alternatively, an oligonucleotide may have at least 90% sequence identity, preferably at least 95% sequence identity, with the exemplary antisense targeting sequences described herein.

[0077] An oligonucleotide "specifically hybridizes" to a target polynucleotide when the oligomer hybridizes to the target under physiological conditions with a Tm substantially above 45°C, preferably at least 50°C, typically 60°C to 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 a complementary polynucleotide. Again, such hybridization can occur with "near" or "substantial" complementarity of the antisense oligomer to the target sequence, as well as exact complementarity.

[0078] A "nuclease-resistant" oligomeric molecule (oligomer) refers to one whose backbone, in either unhybridized or hybridized form, is substantially resistant to nuclease cleavage by common extracellular and intracellular nucleases in the body, i.e., the oligomer exhibits little or no nuclease cleavage under normal nuclease conditions in the body to which the oligomer is exposed.

[0079] 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, either as a single dose or as part of a series of doses, that is effective to produce the desired therapeutic effect. For antisense oligonucleotides, this effect is typically achieved by inhibiting translation or natural splice processing of a selected target sequence. An "effective amount" targeting an UNC13A cryptic exon splice variant mRNA also relates to an amount effective to modulate the expression of an UNC13A cryptic exon splice variant protein.

[0080] The term "inhibit" or "inhibitor" refers to a direct or indirect alteration, interference, reduction, down-regulation, blocking, suppression, abrogation, or degradation in 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, down-regulation, blocking, suppression, abrogation, or degradation is statistically, biologically, or clinically significant. The term "inhibit" or "inhibitor" includes gene "knockout" and gene "knockdown" methods, such as by chromosome editing.

[0081] "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, for example, but is not limited to, the administration of a pharmaceutical composition, and may be administered prophylactically or after the initiation 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, among others, as described herein.

[0082] Also included is "prophylactic" treatment, which may be directed to 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 "prophylaxis" does not necessarily indicate a complete eradication, cure, or prevention of the disease or condition, or the symptoms associated therewith.

[0083] I. UNC13A cryptic exon splice variants In one aspect, the present disclosure provides novel UNC13A-targeting oligonucleotides that reduce cryptic splice variants, including the cryptic exons between exons 20 and 21. These cryptic exons are absent in wild-type UNC13A from neuronal nuclei and in none of the known isoforms of UNC13A. The cryptic exons are derived from introns 20-21 of the UNC13A gene (SEQ ID NO: 4). TDP-43 depletion introduces two alternative 3' splice acceptors in introns 20-21, one at chr19:17642591 (Δψ = 0.05184) and the other at chr19:17642541 (Δψ = 0.48865). An alternative 5' splice donor is also introduced at chr19:17642414 (Δψ = 0.772). The Chr19:176425913' splicing acceptor, which is more frequently used than the Chr19:176425413' splicing acceptor, and an alternative 5' splicing donor result in a 128 bp cryptic exon ("cryptic exon #1") having the nucleotide sequence set forth in SEQ ID NO:5. The UNC13A cryptic exon #1 variant contains the nucleotide sequence set forth in SEQ ID NO:7 and encodes a protein comprising the amino acid sequence set forth in SEQ ID NO:8. The Chr19:176425913' splicing acceptor and an alternative 5' splicing donor result in a 179 bp cryptic exon ("cryptic exon #2") having the nucleotide sequence set forth in SEQ ID NO:6. The UNC13A cryptic exon #2 variant contains the nucleotide sequence set forth in SEQ ID NO:9 and encodes a protein comprising the amino acid sequence set forth in SEQ ID NO:10.

[0084] The expression level of the UNC13A cryptic exon #1 splice variant is significantly increased in the prefrontal cortex of patients with frontotemporal lobar degeneration with TDP-43 inclusion (FTLD-TDP) compared to normal controls. The UNC13A cryptic exon #1 splice variant has also been detected in disease-related tissues of ALS patients. In embodiments, the expression of UNC13A cryptic splice variant #1 or UNC13A cryptic splice variant #2 may be used as a biomarker to identify subjects with TDP-43 protein abnormalities, such as FTLD or ALS.

[0085] When TDP-43 is depleted from the nucleus and accumulates in the cytoplasm, it is phosphorylated. Hyperphosphorylated TDP-43 (pTDP-43) is a key feature of the pathology of TDP-43 proteinopathy. The UNC13A cryptic exon #1 splice variant is strongly associated with phosphorylated TDP-43 levels in FTD / ALS patients. In embodiments, the expression of UNC13A cryptic splice variant #1 or UNC13A cryptic splice variant #2 may be used as a biomarker for phosphorylated TDP-43 levels in subjects.

[0086] Several genetic variants in introns 20–21 of UNC13A have been identified that promote UNC13A cryptic exon inclusion upon TDP-43 depletion. Examples of such variants include 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 CATC 0–2 repeat → 3–5 CATC repeat), and rs62121687 (hg38 chr19:17,642,351 C → A). Furthermore, UNC13A variants that increase cryptic exon inclusion are associated with decreased survival in FTD-ALS patients. In embodiments, identification of a genetic mutation in introns 20-21 of UNC13A in a subject may be used as a biomarker for cryptic exon inclusion in UNC13A. In embodiments, identification of a genetic mutation in introns 20-21 of UNC13A in a subject with a TDP-43 proteinopathy (e.g., FTD, ALS) may be used as a biomarker for decreased survival. [Table 1-1] [Table 1-2] [Table 1-3] [Table 1-4] [Table 1-5] [Table 1-6] [Table 1-7] [Table 1-8] [Table 1-9] [Table 1-10] [Table 1-11] [Table 1-12] [Table 1-13] [Table 1-14] [Table 1-15] [Table 1-16] [Table 1-17] [Table 1-18] [Table 1-19] [Table 1-20] [Table 1-21] [Table 1-22]

[0087] II. UNC13A splice-blocking antisense oligonucleotides In some embodiments, provided herein are antisense oligonucleotides targeting an UNC13A cryptic exon. In embodiments, the cryptic exon is obtained from introns 20-21 of the UNC13A gene. In embodiments, the cryptic exon comprises SEQ ID NO: 5 or SEQ ID NO: 6. In embodiments, the UNC13 cryptic exon splice variant comprises the polynucleotide sequence of SEQ ID NO: 7 or SEQ ID NO: 9. In embodiments, the UNC13A cryptic exon splice 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 splice variant.

[0088] In embodiments, 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 embodiments, the inhibitory nucleic acid comprises an antisense oligonucleotide complementary to the exon 20 splice donor site region in the preprocessed mRNA encoding UNC13A, the cryptic exon splice acceptor site region in the preprocessed mRNA encoding UNC13A, the cryptic exon splice donor site region in the preprocessed mRNA encoding UNC13A, or the exon 21 splice acceptor site region in the preprocessed mRNA encoding UNC13A. In embodiments, the exon 20 splice donor site region comprises or consists of SEQ ID NO: 800. In embodiments, the cryptic exon splice acceptor site region comprises or consists of SEQ ID NO: 801. In embodiments, the cryptic exon splice donor site region comprises or consists of SEQ ID NO: 802. In an embodiment, the exon 21 splice acceptor site comprises or consists of SEQ ID NO:803.

[0089] In embodiments, the antisense oligonucleotides selectively inhibit the expression or activity of UNC13A cryptic exon splice variants spanning full-length UNC13A (wild-type) or other variants (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). [Table 2A-1] [Table 2A-2] [Table 2A-3] [Table 2A-4] [Table 2A-5] [Table 2A-6] [Table 2A-7] [Table 2A-8] [Table 2A-9] [Table 2A-10] [Table 2A-11] [Table 2A-12] [Table 2A-13] [Table 2A-14]

Table 2A-15

Table 2A-16

Table 2A-17

Table 2A-18

Table 2A-19

Table 2A-20

Table 2A-21

Table 2A-22

Table 2A-23

Table 2B-1

Table 2B-2

Table 2B-3

Table 2B-4

Table 2B-5

Table 2B-6

Table 2B-7

Table 2B-8

Table 2B-9

Table 2B-10

Table 2B-11

Table 2B-12

Table 2B-13

Table 2B-14

Table 2B-15

Table 2B-16

Table 2B-17

Table 2B-18

Table 2B-19

Table 2B-20

Table 2B-21

Table 2B-22

[0090] The antisense oligonucleotides (ASOs) provided herein are alternatively referred to by the sequence ID numbers and ASO numbers provided in Tables 2A and 2B. The ASO numbers MTx_ASO_(number) and the number alone are used interchangeably. For example, MTx_ASO_280 is alternatively referred to as ASO280. In some cases, the ASO number refers to the region targeted by the respective ASO. For example, MTx_ASO_Region_4_Child_1 (SEQ ID NO: 961) targets region 4 of the cryptic exon (SEQ ID NO: 703) as described herein.

[0091] In some embodiments, the antisense oligonucleotide is complementary to a cryptic exon of UNC13 mRNA. In some embodiments, the antisense oligonucleotide comprises or consists of a sequence set forth in SEQ ID NOs: 20-40. In some embodiments, the antisense oligonucleotide is about 15-50 bases in length, preferably 16, 17, 18, 19, 20, 21, 22, 23, 24, or 25 bases. In some embodiments, the antisense oligonucleotide comprises a sequence 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 oligonucleotides block the inclusion of cryptic exons through steric blocking of either the cryptic 5' donor site splice or the 3' cryptic acceptor site. The antisense oligonucleotides may block the inclusion of cryptic exons by modulating the rate of RNA polymerase II. The antisense oligonucleotides may block the inclusion of cryptic exons by modifying the structure of mRNA. The antisense oligonucleotides may block the inclusion of cryptic exons through other known or unknown mechanisms that may block the inclusion of cryptic exons. In some embodiments, the antisense oligonucleotides increase UNC13A protein expression. In some embodiments, the antisense oligonucleotides reduce the inclusion of UNC13A cryptic exons.

[0092] In some embodiments, the antisense oligonucleotide is complementary to a cryptic exon of UNC13 mRNA. In some embodiments, the antisense oligonucleotide comprises or consists of a sequence set forth in SEQ ID NOs: 20-355 or 810-957. In some embodiments, the antisense oligonucleotide is about 15-50 bases in length, preferably 16, 17, 18, 19, 20, 21, 22, 23, 24, or 25 bases. In some embodiments, the antisense oligonucleotide comprises a sequence at least 80%, 85%, 90%, 95%, 97%, or 100% identical to any one of SEQ ID NOs: 20-355 or 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 oligonucleotides block the inclusion of cryptic exons through steric blocking of either the cryptic 5' donor site splice or the 3' cryptic acceptor site. The antisense oligonucleotides may block the inclusion of cryptic exons by modulating the rate of RNA polymerase II. The antisense oligonucleotides may block the inclusion of cryptic exons by modifying the structure of mRNA. The antisense oligonucleotides may block the inclusion of cryptic exons through other known or unknown mechanisms that may block the inclusion of cryptic exons. In some embodiments, the antisense oligonucleotides increase UNC13A protein expression. In some embodiments, the antisense oligonucleotides reduce the inclusion of UNC13A cryptic exons.

[0093] In some embodiments, the antisense oligonucleotide is complementary to a cryptic exon of UNC13 mRNA. In some embodiments, the antisense oligonucleotide comprises or consists of the sequence set forth in SEQ ID NOs: 20-355, 400-638, or 810-957. In some embodiments, the antisense oligonucleotide is about 15-50 bases in length, preferably 16, 17, 18, 19, 20, 21, 22, 23, 24, 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, 400-638, or 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 an LNA antisense oligonucleotide. In some embodiments, the antisense oligonucleotide blocks the inclusion of cryptic exons through steric blocking of either a cryptic 5' donor site splice or a 3' cryptic acceptor site. The antisense oligonucleotide may block the inclusion of cryptic exons by modulating the rate of RNA polymerase II. The antisense oligonucleotide may block the inclusion of cryptic exons by modifying the structure of mRNA. The antisense oligonucleotide may block the inclusion of cryptic exons through other known or unknown mechanisms that can block the inclusion of cryptic exons. In some embodiments, the antisense oligonucleotide increases UNC13A protein expression. In some embodiments, the antisense oligonucleotide reduces the inclusion of UNC13A cryptic exons.

[0094] In some embodiments, the antisense oligonucleotide is complementary to a cryptic exon of UNC13 mRNA. In some embodiments, the antisense oligonucleotide comprises or consists of the sequence set forth in SEQ ID NOs: 20-355, 400-638, 810-959, or 961-1332. In some embodiments, the antisense oligonucleotide is about 13-50 bases in length, e.g., 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: 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 cryptic exons through steric blockage of either a cryptic 5' donor site splice or a 3' cryptic acceptor site. The antisense oligonucleotide may block the inclusion of cryptic exons by modulating the rate of RNA polymerase II. The antisense oligonucleotide may block the inclusion of cryptic exons by modifying the structure of the mRNA. The antisense oligonucleotide may block the inclusion of cryptic exons through other known or unknown mechanisms that can block the inclusion of cryptic exons. In some embodiments, the antisense oligonucleotide increases UNC13A protein expression. In some embodiments, the antisense oligonucleotide reduces the inclusion of UNC13A cryptic exons.

[0095] 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 sequence set forth in SEQ ID NO: 20 or SEQ ID NO: 38. In some embodiments, the antisense oligonucleotide is about 15-50 bases in length, preferably 16, 17, 18, 19, 20, 21, 22, 23, 24, or 25 bases. In some embodiments, the antisense oligonucleotide comprises a sequence at least 80%, 85%, 90%, 95%, 97%, or 100% identical to any one of 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 cryptic exons through steric blocking of cryptic 5' donor site splices. The antisense oligonucleotide may block the inclusion of cryptic exons by modulating the rate of RNA polymerase II. The antisense oligonucleotide may block the inclusion of cryptic exons by modifying the structure of mRNA. The antisense oligonucleotide may block the inclusion of cryptic exons by other known or unknown mechanisms that can block the inclusion of cryptic exons. In some embodiments, the antisense oligonucleotide increases UNC13A protein expression. In some embodiments, the antisense oligonucleotide reduces the inclusion of UNC13A cryptic exons.

[0096] 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 sequence set forth in SEQ ID NOs: 41-55 or 810-825. In some embodiments, the antisense oligonucleotide is about 15-50 bases in length, preferably 16, 17, 18, 19, 20, 21, 22, 23, 24, or 25 bases. In some embodiments, the antisense oligonucleotide comprises or consists of a sequence at least 80%, 85%, 90%, 95%, 97%, or 100% identical to any one of SEQ ID NOs: 41-55 or 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 an LNA antisense oligonucleotide. In some embodiments, the antisense oligonucleotide blocks the inclusion of cryptic exons through steric blocking of cryptic 5' donor site splices. The antisense oligonucleotide may block the inclusion of cryptic exons by modulating the rate of RNA polymerase II. The antisense oligonucleotide may block the inclusion of cryptic exons by modifying the structure of mRNA. The antisense oligonucleotide may block the inclusion of cryptic exons by other known or unknown mechanisms that can block the inclusion of cryptic exons. In some embodiments, the antisense oligonucleotide increases UNC13A protein expression. In some embodiments, the antisense oligonucleotide reduces the inclusion of UNC13A cryptic exons.

[0097] 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 sequence set forth in SEQ ID NOs: 311-325 or 910-929. In some embodiments, the antisense oligonucleotide is about 15-50 bases in length, preferably 16, 17, 18, 19, 20, 21, 22, 23, 24, or 25 bases. In some embodiments, the antisense oligonucleotide comprises or consists of a sequence at least 80%, 85%, 90%, 95%, 97%, or 100% identical to any one of SEQ ID NOs: 311-325 or 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 an LNA antisense oligonucleotide. In some embodiments, the antisense oligonucleotide blocks the inclusion of cryptic exons through steric blocking of cryptic 5' donor site splices. The antisense oligonucleotide may block the inclusion of cryptic exons by modulating the rate of RNA polymerase II. The antisense oligonucleotide may block the inclusion of cryptic exons by modifying the structure of mRNA. The antisense oligonucleotide may block the inclusion of cryptic exons by other known or unknown mechanisms that can block the inclusion of cryptic exons. In some embodiments, the antisense oligonucleotide increases UNC13A protein expression. In some embodiments, the antisense oligonucleotide reduces the inclusion of UNC13A cryptic exons.

[0098] In some embodiments, the antisense oligonucleotide is complementary to the middle region of a cryptic exon of UNC13 mRNA located between the 5' and 3' splice sites. 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 sequence set forth in SEQ ID NOs: 34-37. In some embodiments, the antisense oligonucleotide is about 15-50 bases in length, preferably 16, 17, 18, 19, 20, 21, 22, 23, 24, or 25 bases in length. In some embodiments, the antisense oligonucleotide comprises or consists of a sequence 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 an LNA antisense oligonucleotide. In some embodiments, the antisense oligonucleotide blocks the inclusion of cryptic exons through steric blocking of either a cryptic 5' donor site splice or a 3' cryptic acceptor site. The antisense oligonucleotide may block the inclusion of cryptic exons by modulating the rate of RNA polymerase II. The antisense oligonucleotide may block the inclusion of cryptic exons by modifying the structure of mRNA. The antisense oligonucleotide may block the inclusion of cryptic exons through other known or unknown mechanisms that can block the inclusion of cryptic exons. In some embodiments, the antisense oligonucleotide increases UNC13A protein expression. In some embodiments, the antisense oligonucleotide reduces the inclusion of UNC13A cryptic exons.

[0099] In some embodiments, the antisense oligonucleotide is complementary to the middle region of a cryptic exon of UNC13 mRNA located between the 5' and 3' splice sites. 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 sequence set forth in SEQ ID NOs: 251-265 or 877-890. In some embodiments, the antisense oligonucleotide is about 15-50 bases in length, preferably 16, 17, 18, 19, 20, 21, 22, 23, 24, or 25 bases. In some embodiments, the antisense oligonucleotide comprises a sequence at least 80%, 85%, 90%, 95%, 97%, or 100% identical to any one of SEQ ID NOs: 251-265 or 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 an LNA antisense oligonucleotide. In some embodiments, the antisense oligonucleotide blocks the inclusion of cryptic exons through steric blocking of either a cryptic 5' donor site splice or a 3' cryptic acceptor site. The antisense oligonucleotide may block the inclusion of cryptic exons by modulating the rate of RNA polymerase II. The antisense oligonucleotide may block the inclusion of cryptic exons by modifying the structure of mRNA. The antisense oligonucleotide may block the inclusion of cryptic exons through other known or unknown mechanisms that can block the inclusion of cryptic exons. In some embodiments, the antisense oligonucleotide increases UNC13A protein expression. In some embodiments, the antisense oligonucleotide reduces the inclusion of UNC13A cryptic exons.

[0100] In some embodiments, the antisense oligonucleotide is complementary to the middle region of a cryptic exon of UNC13 mRNA located between the 5' and 3' splice sites. 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 sequence set forth in SEQ ID NOs: 266-280 or 891-894. In some embodiments, the antisense oligonucleotide is about 15-50 bases in length, preferably 16, 17, 18, 19, 20, 21, 22, 23, 24, or 25 bases. In some embodiments, the antisense oligonucleotide comprises a sequence at least 80%, 85%, 90%, 95%, 97%, or 100% identical to any one of SEQ ID NOs: 266-280 or 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 an LNA antisense oligonucleotide. In some embodiments, the antisense oligonucleotide blocks the inclusion of cryptic exons through steric blocking of either a cryptic 5' donor site splice or a 3' cryptic acceptor site. The antisense oligonucleotide may block the inclusion of cryptic exons by modulating the rate of RNA polymerase II. The antisense oligonucleotide may block the inclusion of cryptic exons by modifying the structure of mRNA. The antisense oligonucleotide may block the inclusion of cryptic exons through other known or unknown mechanisms that can block the inclusion of cryptic exons. In some embodiments, the antisense oligonucleotide increases UNC13A protein expression. In some embodiments, the antisense oligonucleotide reduces the inclusion of UNC13A cryptic exons.

[0101] In some embodiments, the antisense oligonucleotide is complementary to the middle region of a cryptic exon of UNC13 mRNA located between the 5' and 3' splice sites. 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 sequence set forth in SEQ ID NOs: 281-295 or 895-899. In some embodiments, the antisense oligonucleotide is about 15-50 bases in length, preferably 16, 17, 18, 19, 20, 21, 22, 23, 24, or 25 bases. In some embodiments, the antisense oligonucleotide comprises a sequence at least 80%, 85%, 90%, 95%, 97%, or 100% identical to any one of SEQ ID NOs: 281-295 or 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 an LNA antisense oligonucleotide. In some embodiments, the antisense oligonucleotide blocks the inclusion of cryptic exons through steric blocking of either a cryptic 5' donor site splice or a 3' cryptic acceptor site. The antisense oligonucleotide may block the inclusion of cryptic exons by modulating the rate of RNA polymerase II. The antisense oligonucleotide may block the inclusion of cryptic exons by modifying the structure of mRNA. The antisense oligonucleotide may block the inclusion of cryptic exons through other known or unknown mechanisms that can block the inclusion of cryptic exons. In some embodiments, the antisense oligonucleotide increases UNC13A protein expression. In some embodiments, the antisense oligonucleotide reduces the inclusion of UNC13A cryptic exons.

[0102] In some embodiments, the antisense oligonucleotide is complementary to the middle region of a cryptic exon of UNC13 mRNA located between the 5' and 3' splice sites. 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 sequence set forth in SEQ ID NOs: 296-310 or 900-909. In some embodiments, the antisense oligonucleotide is about 15-50 bases in length, preferably 16, 17, 18, 19, 20, 21, 22, 23, 24, or 25 bases. In some embodiments, the antisense oligonucleotide comprises a sequence at least 80%, 85%, 90%, 95%, 97%, or 100% identical to any one of SEQ ID NOs: 296-310 or 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 cryptic exons through steric blocking of either a cryptic 5' donor site splice or a 3' cryptic acceptor site. The antisense oligonucleotide may block the inclusion of cryptic exons by modulating the rate of RNA polymerase II. The antisense oligonucleotide may block the inclusion of cryptic exons by modifying the structure of mRNA. The antisense oligonucleotide may block the inclusion of cryptic exons through other known or unknown mechanisms that can block the inclusion of cryptic exons. In some embodiments, the antisense oligonucleotide increases UNC13A protein expression. In some embodiments, the antisense oligonucleotide reduces the inclusion of UNC13A cryptic exons.

[0103] In some embodiments, the antisense oligonucleotide is complementary to the middle region of a cryptic exon of UNC13 mRNA located between the 5' and 3' splice sites. 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 set forth in SEQ ID NO: 40. In some embodiments, the antisense oligonucleotide is about 15-50 bases in length, preferably 16, 17, 18, 19, 20, 21, 22, 23, 24, or 25 bases in length. In some embodiments, the antisense oligonucleotide comprises a sequence at least 80%, 85%, 90%, 95%, 97%, or 100% identical to any one of 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 an LNA antisense oligonucleotide. In some embodiments, the antisense oligonucleotide blocks the inclusion of cryptic exons through steric blocking of either a cryptic 5' donor site splice or a 3' cryptic acceptor site. The antisense oligonucleotide may block the inclusion of cryptic exons by modulating the rate of RNA polymerase II. The antisense oligonucleotide may block the inclusion of cryptic exons by modifying the structure of mRNA. The antisense oligonucleotide may block the inclusion of cryptic exons through other known or unknown mechanisms that can block the inclusion of cryptic exons. In some embodiments, the antisense oligonucleotide increases UNC13A protein expression. In some embodiments, the antisense oligonucleotide reduces the inclusion of UNC13A cryptic exons.

[0104] In some embodiments, the antisense oligonucleotide is complementary to the middle region of a cryptic exon of UNC13 mRNA located between the 5' and 3' splice sites. 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 set forth in SEQ ID NOs: 341-355 or 934-957. In some embodiments, the antisense oligonucleotide is about 15-50 bases in length, preferably 16, 17, 18, 19, 20, 21, 22, 23, 24, or 25 bases. In some embodiments, the antisense oligonucleotide comprises a sequence at least 80%, 85%, 90%, 95%, 97%, or 100% identical to any one of SEQ ID NOs: 341-355 or 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 cryptic exons through steric blocking of either a cryptic 5' donor site splice or a 3' cryptic acceptor site. The antisense oligonucleotide may block the inclusion of cryptic exons by modulating the rate of RNA polymerase II. The antisense oligonucleotide may block the inclusion of cryptic exons by modifying the structure of mRNA. The antisense oligonucleotide may block the inclusion of cryptic exons through other known or unknown mechanisms that can block the inclusion of cryptic exons. In some embodiments, the antisense oligonucleotide increases UNC13A protein expression.In some embodiments, the antisense oligonucleotide reduces the inclusion of an UNC13A cryptic exon.

[0105] In some embodiments, the antisense oligonucleotide is complementary to the middle region of a cryptic exon of UNC13 mRNA located between the 5' and 3' splice sites. 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 sequence set forth in SEQ ID NOs: 1116-1332. In some embodiments, the antisense oligonucleotide is about 13-50 bases in length, e.g., 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, or 25 bases. In some embodiments, the antisense oligonucleotide comprises a sequence 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 an LNA antisense oligonucleotide. In some embodiments, the antisense oligonucleotide blocks the inclusion of cryptic exons through steric blocking of either a cryptic 5' donor site splice or a 3' cryptic acceptor site. The antisense oligonucleotide may block the inclusion of cryptic exons by modulating the rate of RNA polymerase II. The antisense oligonucleotide may block the inclusion of cryptic exons by modifying the structure of the mRNA. The antisense oligonucleotide may block the inclusion of cryptic exons through other known or unknown mechanisms that can block the inclusion of cryptic exons. 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 an UNC13A cryptic exon.

[0106] In some embodiments, the antisense oligonucleotide is complementary to the 3' end of a 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 sequence set forth in SEQ ID NOs: 30-33. In some embodiments, the antisense oligonucleotide is about 15-50 bases in length, preferably 16, 17, 18, 19, 20, 21, 22, 23, 24, or 25 bases. In some embodiments, the antisense oligonucleotide comprises a sequence 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 cryptic exon inclusion through steric blocking of the 3' cryptic acceptor site. The antisense oligonucleotide may block cryptic exon inclusion by modulating the rate of RNA polymerase II. The antisense oligonucleotide may block cryptic exon inclusion by modifying the structure of the mRNA. The antisense oligonucleotide may block cryptic exon inclusion by other known or unknown mechanisms that can block cryptic exon inclusion. In some embodiments, the antisense oligonucleotide increases UNC13A protein expression. In some embodiments, the antisense oligonucleotide reduces UNC13A cryptic exon inclusion.

[0107] In some embodiments, the antisense oligonucleotide is complementary to the 3' end of a 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 sequence set forth in SEQ ID NOs: 191-205 or 851-860. In some embodiments, the antisense oligonucleotide is about 15-50 bases in length, preferably 16, 17, 18, 19, 20, 21, 22, 23, 24, or 25 bases. In some embodiments, the antisense oligonucleotide comprises a sequence at least 80%, 85%, 90%, 95%, 97%, or 100% identical to any one of SEQ ID NOs: 191-205 or 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 cryptic exon inclusion through steric blocking of the 3' cryptic acceptor site. The antisense oligonucleotide may block cryptic exon inclusion by modulating the rate of RNA polymerase II. The antisense oligonucleotide may block cryptic exon inclusion by modifying mRNA structure 3. The antisense oligonucleotide may block cryptic exon inclusion by other known or unknown mechanisms that can block cryptic exon inclusion. In some embodiments, the antisense oligonucleotide increases UNC13A protein expression. In some embodiments, the antisense oligonucleotide reduces UNC13A cryptic exon inclusion.

[0108] In some embodiments, the antisense oligonucleotide is complementary to the 3' end of a 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 sequence set forth in SEQ ID NOs: 206-220 or 861-863. In some embodiments, the antisense oligonucleotide is about 15-50 bases in length, preferably 16, 17, 18, 19, 20, 21, 22, 23, 24, or 25 bases. In some embodiments, the antisense oligonucleotide comprises a sequence at least 80%, 85%, 90%, 95%, 97%, or 100% identical to any one of SEQ ID NOs: 206-220 or 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 an LNA antisense oligonucleotide. In some embodiments, the antisense oligonucleotide blocks cryptic exon inclusion through steric blocking of the 3' cryptic acceptor site. The antisense oligonucleotide may block cryptic exon inclusion by modulating the rate of RNA polymerase II. The antisense oligonucleotide may block cryptic exon inclusion by modifying the structure of the mRNA. The antisense oligonucleotide may block cryptic exon inclusion by other known or unknown mechanisms that can block cryptic exon inclusion. In some embodiments, the antisense oligonucleotide increases UNC13A protein expression. In some embodiments, the antisense oligonucleotide reduces UNC13A cryptic exon inclusion.

[0109] In some embodiments, the antisense oligonucleotide is complementary to the 3' end of a 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 sequence set forth in SEQ ID NOs: 221-234 or 864-866. In some embodiments, the antisense oligonucleotide is about 15-50 bases in length, preferably 16, 17, 18, 19, 20, 21, 22, 23, 24, or 25 bases. In some embodiments, the antisense oligonucleotide comprises a sequence at least 80%, 85%, 90%, 95%, 97%, or 100% identical to any one of SEQ ID NOs: 221-225 or 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 cryptic exon inclusion through steric blocking of the 3' cryptic acceptor site. The antisense oligonucleotide may block cryptic exon inclusion by modulating the rate of RNA polymerase II. The antisense oligonucleotide may block cryptic exon inclusion by modifying the structure of the mRNA. The antisense oligonucleotide may block cryptic exon inclusion by other known or unknown mechanisms that can block cryptic exon inclusion. In some embodiments, the antisense oligonucleotide increases UNC13A protein expression. In some embodiments, the antisense oligonucleotide reduces UNC13A cryptic exon inclusion.

[0110] In some embodiments, the antisense oligonucleotide is complementary to the 3' end of a 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 sequence set forth in SEQ ID NOs: 236-250 or 867-876. In some embodiments, the antisense oligonucleotide is about 15-50 bases in length, preferably 16, 17, 18, 19, 20, 21, 22, 23, 24, or 25 bases. In some embodiments, the antisense oligonucleotide comprises a sequence at least 80%, 85%, 90%, 95%, 97%, or 100% identical to any one of SEQ ID NOs: 236-250 or 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 an LNA antisense oligonucleotide. In some embodiments, the antisense oligonucleotide blocks cryptic exon inclusion through steric blocking of the 3' cryptic acceptor site. The antisense oligonucleotide may block cryptic exon inclusion by modulating the rate of RNA polymerase II. The antisense oligonucleotide may block cryptic exon inclusion by modifying the structure of the mRNA. The antisense oligonucleotide may block cryptic exon inclusion by other known or unknown mechanisms that can block cryptic exon inclusion. In some embodiments, the antisense oligonucleotide increases UNC13A protein expression. In some embodiments, the antisense oligonucleotide reduces UNC13A cryptic exon inclusion.

[0111] In some embodiments, the antisense oligonucleotide is complementary to the 3' end of a 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 set forth in SEQ ID NOs: 21-29 or SEQ ID NO: 39. In some embodiments, the antisense oligonucleotide is about 15-50 bases in length, preferably 16, 17, 18, 19, 20, 21, 22, 23, 24, or 25 bases. In some embodiments, the antisense oligonucleotide comprises a sequence at least 80%, 85%, 90%, 95%, 97%, or 100% identical to any one of SEQ ID NOs: 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 cryptic exon inclusion through steric blocking of the 3' cryptic acceptor site. The antisense oligonucleotide may block cryptic exon inclusion by modulating the rate of RNA polymerase II. The antisense oligonucleotide may block cryptic exon inclusion by modifying the structure of the mRNA. The antisense oligonucleotide may block cryptic exon inclusion by other known or unknown mechanisms that can block cryptic exon inclusion. In some embodiments, the antisense oligonucleotide increases UNC13A protein expression. In some embodiments, the antisense oligonucleotide reduces UNC13A cryptic exon inclusion.

[0112] In some embodiments, the antisense oligonucleotide is complementary to the 3' end of a 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 set forth in SEQ ID NOs: 56-70 or 826-834. In some embodiments, the antisense oligonucleotide is about 15-50 bases in length, preferably 16, 17, 18, 19, 20, 21, 22, 23, 24, or 25 bases. In some embodiments, the antisense oligonucleotide comprises a sequence at least 80%, 85%, 90%, 95%, 97%, or 100% identical to any one of SEQ ID NOs: 56-70 or 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 an LNA antisense oligonucleotide. In some embodiments, the antisense oligonucleotide blocks cryptic exon inclusion through steric blocking of the 3' cryptic acceptor site. The antisense oligonucleotide may block cryptic exon inclusion by modulating the rate of RNA polymerase II. The antisense oligonucleotide may block cryptic exon inclusion by modifying the structure of the mRNA. The antisense oligonucleotide may block cryptic exon inclusion by other known or unknown mechanisms that can block cryptic exon inclusion. In some embodiments, the antisense oligonucleotide increases UNC13A protein expression. In some embodiments, the antisense oligonucleotide reduces UNC13A cryptic exon inclusion.

[0113] In some embodiments, the antisense oligonucleotide is complementary to the 3' end of a 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 set forth in SEQ ID NOs: 71-85 or 835-837. In some embodiments, the antisense oligonucleotide is about 15-50 bases in length, preferably 16, 17, 18, 19, 20, 21, 22, 23, 24, or 25 bases. In some embodiments, the antisense oligonucleotide comprises a sequence at least 80%, 85%, 90%, 95%, 97%, or 100% identical to any one of SEQ ID NOs: 71-85 or 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 an LNA antisense oligonucleotide. In some embodiments, the antisense oligonucleotide blocks cryptic exon inclusion through steric blocking of the 3' cryptic acceptor site. The antisense oligonucleotide may block cryptic exon inclusion by modulating the rate of RNA polymerase II. The antisense oligonucleotide may block cryptic exon inclusion by modifying the structure of the mRNA. The antisense oligonucleotide may block cryptic exon inclusion by other known or unknown mechanisms that can block cryptic exon inclusion. In some embodiments, the antisense oligonucleotide increases UNC13A protein expression. In some embodiments, the antisense oligonucleotide reduces UNC13A cryptic exon inclusion.

[0114] In some embodiments, the antisense oligonucleotide is complementary to the 3' end of a 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 a sequence set forth in SEQ ID NOs: 86-100. In some embodiments, the antisense oligonucleotide is about 15-50 bases in length, preferably 16, 17, 18, 19, 20, 21, 22, 23, 24, or 25 bases. In some embodiments, the antisense oligonucleotide comprises a sequence 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 an LNA antisense oligonucleotide. In some embodiments, the antisense oligonucleotide blocks cryptic exon inclusion through steric blocking of the 3' cryptic acceptor site. The antisense oligonucleotide may block cryptic exon inclusion by modulating the rate of RNA polymerase II. The antisense oligonucleotide may block cryptic exon inclusion by modifying the structure of the mRNA. The antisense oligonucleotide may block cryptic exon inclusion by other known or unknown mechanisms that can block cryptic exon inclusion. In some embodiments, the antisense oligonucleotide increases UNC13A protein expression. In some embodiments, the antisense oligonucleotide reduces UNC13A cryptic exon inclusion.

[0115] In some embodiments, the antisense oligonucleotide is complementary to the 3' end of a 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 a sequence set forth in SEQ ID NOs: 101-115. In some embodiments, the antisense oligonucleotide is about 15-50 bases in length, preferably 16, 17, 18, 19, 20, 21, 22, 23, 24, or 25 bases. In some embodiments, the antisense oligonucleotide comprises a sequence 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 an LNA antisense oligonucleotide. In some embodiments, the antisense oligonucleotide blocks cryptic exon inclusion through steric blocking of the 3' cryptic acceptor site. The antisense oligonucleotide may block cryptic exon inclusion by modulating the rate of RNA polymerase II. The antisense oligonucleotide may block cryptic exon inclusion by modifying the structure of the mRNA. The antisense oligonucleotide may block cryptic exon inclusion by other known or unknown mechanisms that can block cryptic exon inclusion. In some embodiments, the antisense oligonucleotide increases UNC13A protein expression. In some embodiments, the antisense oligonucleotide reduces UNC13A cryptic exon inclusion.

[0116] In some embodiments, the antisense oligonucleotide is complementary to the 3' end of a 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 a sequence set forth in SEQ ID NOs: 116-130. In some embodiments, the antisense oligonucleotide is about 15-50 bases in length, preferably 16, 17, 18, 19, 20, 21, 22, 23, 24, or 25 bases. In some embodiments, the antisense oligonucleotide comprises a sequence 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 cryptic exon inclusion through steric blocking of the 3' cryptic acceptor site. The antisense oligonucleotide may block cryptic exon inclusion by modulating the rate of RNA polymerase II. The antisense oligonucleotide may block cryptic exon inclusion by modifying the structure of the mRNA. The antisense oligonucleotide may block cryptic exon inclusion by other known or unknown mechanisms that can block cryptic exon inclusion. In some embodiments, the antisense oligonucleotide increases UNC13A protein expression. In some embodiments, the antisense oligonucleotide reduces UNC13A cryptic exon inclusion.

[0117] In some embodiments, the antisense oligonucleotide is complementary to the 3' end of a 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 a sequence set forth in SEQ ID NOs: 131-145. In some embodiments, the antisense oligonucleotide is about 15-50 bases in length, preferably 16, 17, 18, 19, 20, 21, 22, 23, 24, or 25 bases. In some embodiments, the antisense oligonucleotide comprises a sequence 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 an LNA antisense oligonucleotide. In some embodiments, the antisense oligonucleotide blocks cryptic exon inclusion through steric blocking of the 3' cryptic acceptor site. The antisense oligonucleotide may block cryptic exon inclusion by modulating the rate of RNA polymerase II. The antisense oligonucleotide may block cryptic exon inclusion by modifying the structure of the mRNA. The antisense oligonucleotide may block cryptic exon inclusion by other known or unknown mechanisms that can block cryptic exon inclusion. In some embodiments, the antisense oligonucleotide increases UNC13A protein expression. In some embodiments, the antisense oligonucleotide reduces UNC13A cryptic exon inclusion.

[0118] In some embodiments, the antisense oligonucleotide is complementary to the 3' end of a 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 set forth in SEQ ID NOs: 56-70. In some embodiments, the antisense oligonucleotide is about 146-160 bases in length, preferably 16, 17, 18, 19, 20, 21, 22, 23, 24, or 25 bases. In some embodiments, the antisense oligonucleotide comprises a sequence 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 cryptic exon inclusion through steric blocking of the 3' cryptic acceptor site. The antisense oligonucleotide may block cryptic exon inclusion by modulating the rate of RNA polymerase II. The antisense oligonucleotide may block cryptic exon inclusion by modifying the structure of the mRNA. The antisense oligonucleotide may block cryptic exon inclusion by other known or unknown mechanisms that can block cryptic exon inclusion. In some embodiments, the antisense oligonucleotide increases UNC13A protein expression. In some embodiments, the antisense oligonucleotide reduces UNC13A cryptic exon inclusion.

[0119] In some embodiments, the antisense oligonucleotide is complementary to the 3' end of a 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 set forth in SEQ ID NOs: 161-175 or 838-840. In some embodiments, the antisense oligonucleotide is about 15-50 bases in length, preferably 16, 17, 18, 19, 20, 21, 22, 23, 24, or 25 bases. In some embodiments, the antisense oligonucleotide comprises a sequence at least 80%, 85%, 90%, 95%, 97%, or 100% identical to any one of SEQ ID NOs: 161-175 or 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 an LNA antisense oligonucleotide. In some embodiments, the antisense oligonucleotide blocks cryptic exon inclusion through steric blocking of the 3' cryptic acceptor site. The antisense oligonucleotide may block cryptic exon inclusion by modulating the rate of RNA polymerase II. The antisense oligonucleotide may block cryptic exon inclusion by modifying the structure of the mRNA. The antisense oligonucleotide may block cryptic exon inclusion by other known or unknown mechanisms that can block cryptic exon inclusion. In some embodiments, the antisense oligonucleotide increases UNC13A protein expression. In some embodiments, the antisense oligonucleotide reduces UNC13A cryptic exon inclusion.

[0120] In some embodiments, the antisense oligonucleotide is complementary to the 3' end of a 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 set forth in SEQ ID NOs: 176-190 or 841-850. In some embodiments, the antisense oligonucleotide is about 15-50 bases in length, preferably 16, 17, 18, 19, 20, 21, 22, 23, 24, or 25 bases. In some embodiments, the antisense oligonucleotide comprises a sequence at least 80%, 85%, 90%, 95%, 97%, or 100% identical to any one of SEQ ID NOs: 176-190 or 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 an LNA antisense oligonucleotide. In some embodiments, the antisense oligonucleotide blocks cryptic exon inclusion through steric blocking of the 3' cryptic acceptor site. The antisense oligonucleotide may block cryptic exon inclusion by modulating the rate of RNA polymerase II. The antisense oligonucleotide may block cryptic exon inclusion by modifying the structure of the mRNA. The antisense oligonucleotide may block cryptic exon inclusion by other known or unknown mechanisms that can block cryptic exon inclusion. In some embodiments, the antisense oligonucleotide increases UNC13A protein expression. In some embodiments, the antisense oligonucleotide reduces UNC13A cryptic exon inclusion.

[0121] In some embodiments, the antisense oligonucleotide is complementary to the 3' end of a 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 set forth in SEQ ID NOs: 326-340 or 930-933. In some embodiments, the antisense oligonucleotide is about 15-50 bases in length, preferably 16, 17, 18, 19, 20, 21, 22, 23, 24, or 25 bases. In some embodiments, the antisense oligonucleotide comprises a sequence at least 80%, 85%, 90%, 95%, 97%, or 100% identical to any one of SEQ ID NOs: 326-340 or 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 an LNA antisense oligonucleotide. In some embodiments, the antisense oligonucleotide blocks cryptic exon inclusion through steric blocking of the 3' cryptic acceptor site. The antisense oligonucleotide may block cryptic exon inclusion by modulating the rate of RNA polymerase II. The antisense oligonucleotide may block cryptic exon inclusion by modifying the structure of the mRNA. The antisense oligonucleotide may block cryptic exon inclusion by other known or unknown mechanisms that can block cryptic exon inclusion. In some embodiments, the antisense oligonucleotide increases UNC13A protein expression. In some embodiments, the antisense oligonucleotide reduces UNC13A cryptic exon inclusion.

[0122] In some embodiments, the antisense oligonucleotide is complementary to the 3' end of a 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 a sequence set forth in SEQ ID NOs: 961-1115. In some embodiments, the antisense oligonucleotide is about 13-50 bases in length, e.g., 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 polypeptide comprises a sequence 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 cryptic exon inclusion through steric blocking of the 3' cryptic acceptor site. The antisense oligonucleotide may block cryptic exon inclusion by modulating the rate of RNA polymerase II. The antisense oligonucleotide may block cryptic exon inclusion by modifying the structure of the mRNA. The antisense oligonucleotide may block cryptic exon inclusion by other known or unknown mechanisms that can block cryptic exon inclusion. 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 an UNC13A cryptic exon.

[0123] In some embodiments, the antisense oligonucleotide is complementary to the 3' end of a 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 of SEQ ID NOs: 700-703. In some embodiments, the antisense oligonucleotide is complementary to a cryptic exon of UNC13 mRNA. In some embodiments, the antisense oligonucleotide comprises or consists of a sequence set forth in SEQ ID NOs: 400-638. In some embodiments, the antisense oligonucleotide is about 15-50 bases in length, preferably 16, 17, 18, 19, 20, 21, 22, 23, 24, or 25 bases in length. In some embodiments, the antisense oligonucleotide comprises a sequence 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 cryptic exons through steric blocking of either a cryptic 5' donor site splice or a 3' cryptic acceptor site. The antisense oligonucleotide may block the inclusion of cryptic exons by modulating the rate of RNA polymerase II. The antisense oligonucleotide may block the inclusion of cryptic exons by modifying the structure of mRNA. The antisense oligonucleotide may block the inclusion of cryptic exons through other known or unknown mechanisms that can block the inclusion of cryptic exons. In some embodiments, the antisense oligonucleotide increases UNC13A protein expression.In some embodiments, the antisense oligonucleotide reduces the inclusion of an UNC13A cryptic exon.

[0124] In some embodiments, the antisense oligonucleotide has at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 97%, or 100% complementarity to any one of SEQ ID NOs: 5-10 or 700-703. In some embodiments, the antisense oligonucleotide is about 15-50 bases in length, preferably 16, 17, 18, 19, 20, 21, 22, 23, 24, 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 an LNA antisense oligonucleotide. In some embodiments, the antisense oligonucleotide blocks the inclusion of a cryptic exon via steric blocking of either a cryptic 5' donor site splice or a 3' cryptic acceptor site. Antisense oligonucleotides may block the inclusion of cryptic exons by modulating the rate of RNA polymerase II. Antisense oligonucleotides may block the inclusion of cryptic exons by modifying the structure of mRNA. Antisense oligonucleotides may block the inclusion of cryptic exons by other known or unknown mechanisms that may block the inclusion of cryptic exons. In some embodiments, antisense oligonucleotides increase UNC13A protein expression. In some embodiments, antisense oligonucleotides reduce the inclusion of UNC13A cryptic exons.

[0125] In some embodiments, the present specification also provides antisense oligonucleotides that regulate UNC13A splicing, wherein the antisense oligonucleotide has a sequence that is complementary to a sequence in 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.

[0126] In some embodiments, the antisense oligonucleotide is complementary to the sequence set forth 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 to the sequence set forth in SEQ ID NO: 1.

[0127] In some embodiments, the antisense oligonucleotide is complementary to the sequence set forth in any one of SEQ ID NOs: 700-703. In some embodiments, the antisense oligonucleotide is complementary to the sequence set forth 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 to the sequence set forth in SEQ ID NO: 701 or 703. In some embodiments, the antisense oligonucleotide is fully complementary to the sequence set forth in SEQ ID NO: 701 or 703. In some embodiments, the antisense oligonucleotide is complementary to the sequence set forth 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 to the sequence set forth in SEQ ID NO: 700. In some embodiments, the antisense oligonucleotide is fully complementary to the sequence set forth in SEQ ID NO: 700. In some embodiments, the antisense oligonucleotide is complementary to the sequence set forth 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 to the sequence set forth in SEQ ID NO: 701. In some embodiments, the antisense oligonucleotide is fully complementary to the sequence set forth in SEQ ID NO: 701. In some embodiments, the antisense oligonucleotide is complementary to the sequence set forth 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 to the sequence set forth in SEQ ID NO: 702. In some embodiments, the antisense oligonucleotide is fully complementary to the sequence set forth in SEQ ID NO: 702. In some embodiments, the antisense oligonucleotide is complementary to the sequence set forth 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 to the sequence set forth in SEQ ID NO: 703. In some embodiments, the antisense oligonucleotide is fully complementary to the sequence set forth in SEQ ID NO: 703.

[0128] 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 set forth 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 set forth 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 set forth in SEQ ID NO: 703.

[0129] In some cases, the antisense oligonucleotide is complementary to a sequence of the 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, and the UNC13A sequence is uniquely sensitive to antisense oligonucleotide binding. In some cases, regions of the UNC13A mRNA are identified as being more sensitive than other regions. These regions are determined herein to be sensitive to a number of factors, including, but not limited to, antisense oligonucleotide length, composition, and location on the UNC13A mRNA. Minimal regions of the UNC13A mRNA are specifically targeted to maximize antisense oligonucleotide activity in modulating UNC13A mRNA splicing. In some embodiments, these highly responsive minimal regions of the UNC13A mRNA are referred to as hotspots.

[0130] In some embodiments, a minimal region 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 is identified that is 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 inclusion of the UNC13A cryptic exon. In some embodiments, the minimal region is 3-30, 4-26, 5-22, 6-18, 7-14, or 8-10 nucleotides in length. In some embodiments, the minimal region is 30 nucleotides or less, 25 nucleotides or less, 20 nucleotides or less, or 15 nucleotides or less 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 set forth in any one of SEQ ID NOs: 1339-1342, or a portion thereof. In some embodiments, the minimal region comprises the sequence set forth in SEQ ID NO: 1339, or a portion thereof. In some embodiments, the minimal region comprises the sequence set forth in SEQ ID NO: 1340, or a portion thereof. In some embodiments, the minimal region comprises the sequence set forth in SEQ ID NO: 1341, or a portion thereof. In some embodiments, the minimal region comprises the sequence set forth in SEQ ID NO: 1342, or a portion thereof. In some embodiments, combining the subregion, or a portion thereof, with the antisense oligonucleotide results in improved splice regulatory activity. In some embodiments, combining the minimal region or a portion thereof with an antisense oligonucleotide results in increased UNC13A mRNA expression, hi some embodiments, combining the minimal region or a portion thereof with an antisense oligonucleotide results in decreased UNC13A cryptic exon mRNA expression.In some embodiments, binding of the minimal region or a portion thereof with an antisense oligonucleotide results in an increase in UNC13A protein expression. [Table 3]

[0131] In some cases, the antisense oligonucleotide comprises 13 to 30 nucleotides, and the antisense oligonucleotide is complementary to the sequence set forth in any one of SEQ ID NOs: 1339-1342, or a portion thereof. In some embodiments, the antisense oligonucleotide is complementary to the sequence set forth in SEQ ID NO: 1339, or a portion thereof. In some embodiments, the antisense oligonucleotide is complementary to the sequence set forth in SEQ ID NO: 1340, or a portion thereof. In some embodiments, the antisense oligonucleotide is complementary to the sequence set forth in SEQ ID NO: 1341, or a portion thereof. In some embodiments, the antisense oligonucleotide is complementary to the sequence set forth 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 set forth 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 set forth 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 consecutive nucleotides of the sequence set forth 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 set forth 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 set forth 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 set forth 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 set forth 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 set forth in SEQ ID NO: 1342.

[0132] In some embodiments, binding of an antisense oligonucleotide to a sequence set forth in any one of SEQ ID NOs: 1339-1342, or a portion thereof, results in increased UNC13A splice regulatory activity. In some embodiments, binding of an antisense oligonucleotide to a sequence set forth in any one of SEQ ID NOs: 1339-1342, or a portion thereof, results in increased expression of UNC13A mRNA. In some embodiments, expression of UNC13A mRNA is increased in cells by 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, expression of UNC13A mRNA is increased in cells by between about 10% and about 95%, 20% and about 85%, 30% and about 75%, 40% and about 65%, or 50% and about 55% of the level of UNC13A mRNA expressed in wild-type cells. In some embodiments, expression of UNC13A mRNA is increased 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, expression of UNC13A mRNA is increased in cells 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 an antisense oligonucleotide to a sequence or portion set forth in any one of SEQ ID NOs: 1339-1342 results in a decrease in UNC13A cryptic exon mRNA expression. In some embodiments, expression of UNC13A cryptic exon mRNA is decreased in cells by at least 10%, at least 20%, at least 30%, at least 40%, or at least 50%. In some embodiments, expression of UNC13A cryptic exon mRNA is reduced in cells 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 antisense oligonucleotide consists of the sequence set forth 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 set forth in any one of SEQ ID NOs: 1059-1074, 1075-1089, and 1090-1103. In some embodiments, the antisense oligonucleotide consists of the sequence set forth in any one of SEQ ID NOs: 1248-1270, 1271-1292, and 1293-1313.

[0133] In some embodiments, the wild-type cell is from an individual who does not have amyotrophic lateral sclerosis (ALS) or frontotemporal dementia (FTD). In some embodiments, the wild-type cell is a cell that expresses normal levels of TDP-43 protein. In some embodiments, the wild-type cell is a cell that does not produce a transcript of UNC13A that contains a cryptic exon.

[0134] In some aspects, provided herein are antisense oligonucleotides comprising any one of SEQ ID NOs: 20-355, 400-638, 810-959, or 961-1332. In some embodiments, the antisense oligonucleotides comprise or consist of 13-30 nucleotides. In some embodiments, the antisense oligonucleotides comprise or consist of 13-30, 15-28, 17-26, 19-24, or 21-23 nucleotides. In some embodiments, the antisense oligonucleotides comprise or consist of 13-25 nucleotides. In some embodiments, the antisense oligonucleotides comprise 13, 15, 17, 18, 19, 20, 21, 22, or 24 nucleotides. In some embodiments, the antisense oligonucleotides comprise or consist 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 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, the antisense oligonucleotide is provided to a cell, and expression of UNC13A mRNA is increased in the cell by 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, expression of UNC13A mRNA is increased in the cell by between about 10% and about 95%, 20% and about 85%, 30% and about 75%, 40% and about 65%, or 50% and about 55% of the level of UNC13A mRNA expressed in wild-type cells. In some embodiments, expression of UNC13A mRNA is increased in the cells 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, when an antisense oligonucleotide is provided to the cells, expression of UNC13A cryptic exon mRNA is decreased in the cells 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, when an antisense oligonucleotide is provided to the cells, expression of UNC13A protein is increased in the cells by 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, expression of UNC13A protein is increased in cells by between about 10% and about 95%, 20% and about 85%, 30% and about 75%, 40% and about 65%, or 50% and about 55% of the level of UNC13A protein expressed in wild-type cells.In some embodiments, expression of UNC13A protein is increased in cells 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, in which an antisense oligonucleotide is provided to a cell, 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 cell morphology, neurite length, neurotransmitter release, action potentials, neuronal synaptic plasticity, and neuronal longevity. In some embodiments, the antisense oligonucleotide consists of the sequence set forth 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 set forth in any one of SEQ ID NOs: 1059-1074, 1075-1089, and 1090-1103. In some embodiments, the antisense oligonucleotide consists of the sequence set forth 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 oligonucleotides, 2'O-methoxyethyl antisense oligonucleotides, phosphorothioate antisense oligonucleotides, and locked nucleic acid (LNA) antisense oligonucleotides.

[0135] In some embodiments, the antisense oligonucleotide is complementary to the sequence set forth in SEQ ID NO: 1. In some embodiments, the antisense oligonucleotide comprises or consists of the sequence set forth 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 13-30 nucleotides. In some embodiments, the antisense oligonucleotide comprises a modification.

[0136] In some embodiments, the antisense oligonucleotide consists of 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 13-30 nucleotides. In some embodiments, the antisense oligonucleotide comprises a modification.

[0137] In some embodiments, the antisense oligonucleotide comprises 13 nucleobases complementary to the sequence set forth in SEQ ID NO: 1, wherein the antisense oligonucleotide comprises a sequence set forth in any one of SEQ ID NOs: 961-983 and 1116-1145. In some embodiments, the antisense oligonucleotide consists of 13 nucleotides complementary to the sequence set forth in SEQ ID NO: 1, wherein the antisense oligonucleotide consists of a sequence set forth in any one of SEQ ID NOs: 961-983 and 1116-1145. In some embodiments, wherein the antisense oligonucleotide is provided to a cell, expression of UNC13A mRNA is increased in the cell by 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 a wild-type cell. In some embodiments, expression of UNC13A mRNA is increased in cells 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, expression of UNC13A mRNA is increased in cells 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, in which an antisense oligonucleotide is provided to a cell, expression of UNC13A cryptic exon mRNA is reduced in cells 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, in which an antisense oligonucleotide is provided to a cell, expression of UNC13A protein is increased in the cell by 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 a wild-type cell. In some embodiments, expression of UNC13A protein is increased in the cell by between about 10% and about 95%, 20% and about 85%, 30% and about 75%, 40% and about 65%, or 50% and about 55% of the level of UNC13A protein expressed in a wild-type cell.In some embodiments, expression of UNC13A protein is increased in cells 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, in which an antisense oligonucleotide is provided to a cell, 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 cell morphology, neurite length, neurotransmitter release, action potentials, neuronal synaptic plasticity, and neuronal longevity. In some embodiments, the antisense oligonucleotide consists of the sequence set forth 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 set forth in any one of SEQ ID NOs: 1059-1074, 1075-1089, and 1090-1103. In some embodiments, the antisense oligonucleotide consists of the sequence set forth 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 oligonucleotides, 2'O-methoxyethyl antisense oligonucleotides, phosphorothioate antisense oligonucleotides, and locked nucleic acid (LNA) antisense oligonucleotides.

[0138] In some embodiments, the antisense oligonucleotide comprises 15 nucleotides complementary to the sequence set forth in SEQ ID NO: 1, wherein the antisense oligonucleotide comprises a sequence set forth in any one of SEQ ID NOs: 984-1004 and 1146-1173. In some embodiments, the antisense oligonucleotide consists of 15 nucleotides complementary to the sequence set forth in SEQ ID NO: 1, wherein the antisense oligonucleotide consists of a sequence set forth in any one of SEQ ID NOs: 984-1004 and 1146-1173. In some embodiments, wherein the antisense oligonucleotide is provided to a cell, expression of UNC13A mRNA is increased in the cell by 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 a wild-type cell. In some embodiments, expression of UNC13A mRNA is increased in cells 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, expression of UNC13A mRNA is increased in cells 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, in which an antisense oligonucleotide is provided to a cell, expression of UNC13A cryptic exon mRNA is reduced in cells 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, in which an antisense oligonucleotide is provided to a cell, expression of UNC13A protein is increased in the cell by 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 a wild-type cell. In some embodiments, expression of UNC13A protein is increased in the cell by between about 10% and about 95%, 20% and about 85%, 30% and about 75%, 40% and about 65%, or 50% and about 55% of the level of UNC13A protein expressed in a wild-type cell.In some embodiments, expression of UNC13A protein is increased in cells 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, in which an antisense oligonucleotide is provided to a cell, 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 cell morphology, neurite length, neurotransmitter release, action potentials, neuronal synaptic plasticity, and neuronal longevity. In some embodiments, the antisense oligonucleotide consists of the sequence set forth 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 set forth in any one of SEQ ID NOs: 1059-1074, 1075-1089, and 1090-1103. In some embodiments, the antisense oligonucleotide consists of the sequence set forth 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 oligonucleotides, 2'O-methoxyethyl antisense oligonucleotides, phosphorothioate antisense oligonucleotides, and locked nucleic acid (LNA) antisense oligonucleotides.

[0139] In some embodiments, the antisense oligonucleotide comprises 16 nucleotides complementary to the sequence set forth in SEQ ID NO: 1, and the antisense oligonucleotide comprises a sequence set forth in 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 set forth in SEQ ID NO: 1, and the antisense oligonucleotide consists of 16 nucleotides complementary to the sequence set forth in 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 a cell, expression of UNC13A mRNA is increased in the cell by 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 a wild-type cell. In some embodiments, expression of UNC13A mRNA is increased in cells by between about 10% and about 95%, 20% and about 85%, 30% and about 75%, 40% and about 65%, or 50% and about 55% of the level of UNC13A mRNA expressed in wild-type cells.In some embodiments, expression of UNC13A mRNA is increased in the cells 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, when an antisense oligonucleotide is provided to the cells, expression of UNC13A cryptic exon mRNA is decreased in the cells 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, when an antisense oligonucleotide is provided to the cells, expression of UNC13A protein is increased in the cells by 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, expression of UNC13A protein is increased in cells by between about 10% and about 95%, 20% and about 85%, 30% and about 75%, 40% and about 65%, or 50% and about 55% of the level of UNC13A protein expressed in wild-type cells. In some embodiments, expression of UNC13A protein is increased in cells by between about 25% and about 1000%, about 50% and about 800%, about 100% and about 700%, about 200% and about 600%, or about 300% and about 500%. In some embodiments in which an antisense oligonucleotide is provided to a cell, 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 potentials, neuronal synaptic plasticity, and neuronal longevity. In some embodiments, the antisense oligonucleotide consists of the sequence set forth 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 set forth in any one of SEQ ID NOs: 1059-1074, 1075-1089, and 1090-1103.In some embodiments, the antisense oligonucleotide consists of 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 oligonucleotides, 2'O-methoxyethyl antisense oligonucleotides, phosphorothioate antisense oligonucleotides, and locked nucleic acid (LNA) antisense oligonucleotides.

[0140] In some embodiments, the antisense oligonucleotide comprises 17 nucleotides complementary to the sequence set forth in SEQ ID NO: 1, and the antisense oligonucleotide comprises a sequence set forth 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 17 nucleotides complementary to the sequence set forth in SEQ ID NO: 1, and the antisense oligonucleotide consists of any one of the sequences set forth in 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, where an antisense oligonucleotide is provided to a cell, expression of UNC13A mRNA is increased in the cell by 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 a wild-type cell. In some embodiments, expression of UNC13A mRNA is increased in the cell by between about 10% and about 95%, 20% and about 85%, 30% and about 75%, 40% and about 65%, or 50% and about 55% of the level of UNC13A mRNA expressed in a wild-type cell. In some embodiments, expression of UNC13A mRNA is increased in the cell by between about 25% and about 1000%, about 50% and about 800%, about 100% and about 700%, about 200% and about 600%, or about 300% and about 500%.In some embodiments, where an antisense oligonucleotide is provided to a cell, expression of UNC13A cryptic exon mRNA is reduced in the cell 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, expression of UNC13A protein is increased in the cell by 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, expression of UNC13A protein is increased in the cell by between 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, expression of UNC13A protein is increased in cells 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, in which an antisense oligonucleotide is provided to a cell, 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 cell morphology, neurite length, neurotransmitter release, action potentials, neuronal synaptic plasticity, and neuronal longevity. In some embodiments, the antisense oligonucleotide consists of the sequence set forth 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 set forth in any one of SEQ ID NOs: 1059-1074, 1075-1089, and 1090-1103. In some embodiments, the antisense oligonucleotide consists of the sequence set forth 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 oligonucleotides, 2'O-methoxyethyl antisense oligonucleotides, phosphorothioate antisense oligonucleotides, and locked nucleic acid (LNA) antisense oligonucleotides.

[0141] In some embodiments, the antisense oligonucleotide comprises 18 nucleotides complementary to the sequence set forth in SEQ ID NO: 1, and the antisense oligonucleotide comprises any 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, 190, 200, 210, 212, 214, 216, 218, 220, 222, 224, 226, 228, 230, 232, 234, 235, 249, 150, 164, 165, 179, 180, 4, 195, 209, 210, 224, 225, 239, 240, 254, 255, 269, 270, 284, 285, 299, 300, 314, 329, 344, 518 to 638, 817, 818, 845, 855, 871, 882, 904, 958, 959, 960, 1024 to 1041, and 1200 to 1224. In some embodiments, the antisense oligonucleotide consists of 18 nucleotides complementary to the sequence set forth in SEQ ID NO: 1, and the antisense oligonucleotide is selected from the group consisting 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, 190, 200, 210, 212, 214, 216, 218, 220, 222, 224, 226, 228, 230, 232, 234, 235, 249, 150, 164, 165, 179, 180, 191, 219, 225, 232, 233, 234, 235, 249, 150, 164, 165, 179, 180, 192, 235, 246, 247, 248, 250, 252, 254, 256, 258, 260, 262, 264, 265, 270, 272, 274, 275, 276, 278, 279, 280, 281, 282, 283, 284, 285, 286, 287, 288, 289, 300, 301, 302, 303, 304, 305, 4, 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 a cell, expression of UNC13A mRNA is increased in the cell by 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 a wild-type cell. In some embodiments, expression of UNC13A mRNA is increased in cells by between about 10% and about 95%, 20% and about 85%, 30% and about 75%, 40% and about 65%, or 50% and about 55% of the level of UNC13A mRNA expressed in wild-type cells.In some embodiments, expression of UNC13A mRNA is increased in the cells 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, when an antisense oligonucleotide is provided to the cells, expression of UNC13A cryptic exon mRNA is decreased in the cells 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, when an antisense oligonucleotide is provided to the cells, expression of UNC13A protein is increased in the cells by 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, expression of UNC13A protein is increased in cells by between about 10% and about 95%, 20% and about 85%, 30% and about 75%, 40% and about 65%, or 50% and about 55% of the level of UNC13A protein expressed in wild-type cells. In some embodiments, expression of UNC13A protein is increased in cells by between about 25% and about 1000%, about 50% and about 800%, about 100% and about 700%, about 200% and about 600%, or about 300% and about 500%. In some embodiments in which an antisense oligonucleotide is provided to a cell, 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 potentials, neuronal synaptic plasticity, and neuronal longevity. In some embodiments, the antisense oligonucleotide consists of the sequence set forth 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 set forth in any one of SEQ ID NOs: 1059-1074, 1075-1089, and 1090-1103.In some embodiments, the antisense oligonucleotide consists of 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 oligonucleotides, 2'O-methoxyethyl antisense oligonucleotides, phosphorothioate antisense oligonucleotides, and locked nucleic acid (LNA) antisense oligonucleotides.

[0142] In some embodiments, the antisense oligonucleotide comprises 19 nucleotides complementary to the sequence set forth in SEQ ID NO: 1, and the antisense oligonucleotide comprises a sequence set forth 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 19 nucleotides complementary to the sequence set forth in SEQ ID NO: 1, and the antisense oligonucleotide consists of the sequence set forth 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, where an antisense oligonucleotide is provided to a cell, expression of UNC13A mRNA is increased in the cell by 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 a wild-type cell. In some embodiments, expression of UNC13A mRNA is increased in the cell by between about 10% and about 95%, 20% and about 85%, 30% and about 75%, 40% and about 65%, or 50% and about 55% of the level of UNC13A mRNA expressed in a wild-type cell. In some embodiments, expression of UNC13A mRNA is increased in the cell by between about 25% and about 1000%, about 50% and about 800%, about 100% and about 700%, about 200% and about 600%, or about 300% and about 500%. In some embodiments in which an antisense oligonucleotide is provided to a cell, expression of UNC13A cryptic exon mRNA is reduced in the cell 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, in which an antisense oligonucleotide is provided to a cell, expression of UNC13A protein is increased in the cell by 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 a wild-type cell. In some embodiments, expression of UNC13A protein is increased in the cell by between about 10% and about 95%, 20% and about 85%, 30% and about 75%, 40% and about 65%, or 50% and about 55% of the level of UNC13A protein expressed in a wild-type cell. In some embodiments, expression of UNC13A protein is increased in the cell by between about 25% and about 1000%, about 50% and about 800%, about 100% and about 700%, about 200% and about 600%, or about 300% and about 500%. In some embodiments, in which an antisense oligonucleotide is provided to a cell, 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 cell morphology, neurite length, neurotransmitter release, action potentials, neuronal synaptic plasticity, and neuronal longevity. In some embodiments, the antisense oligonucleotide consists of the sequence set forth 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 set forth in any one of SEQ ID NOs: 1059-1074, 1075-1089, and 1090-1103. In some embodiments, the antisense oligonucleotide consists of the sequence set forth 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 oligonucleotides, 2'O-methoxyethyl antisense oligonucleotides, phosphorothioate antisense oligonucleotides, and locked nucleic acid (LNA) antisense oligonucleotides.

[0143] In some embodiments, the antisense oligonucleotide comprises 20 nucleotides complementary to the sequence set forth in SEQ ID NO: 1, and the antisense oligonucleotide comprises a sequence set forth 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 20 nucleotides complementary to the sequence set forth in SEQ ID NO: 1, and the antisense oligonucleotide consists of the sequence set forth 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, where an antisense oligonucleotide is provided to a cell, expression of UNC13A mRNA is increased in the cell by 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 a wild-type cell. In some embodiments, expression of UNC13A mRNA is increased in the cell by between about 10% and about 95%, 20% and about 85%, 30% and about 75%, 40% and about 65%, or 50% and about 55% of the level of UNC13A mRNA expressed in a wild-type cell. In some embodiments, expression of UNC13A mRNA is increased in the cell by between about 25% and about 1000%, about 50% and about 800%, about 100% and about 700%, about 200% and about 600%, or about 300% and about 500%.In some embodiments, where an antisense oligonucleotide is provided to a cell, expression of UNC13A cryptic exon mRNA is reduced in the cell 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, expression of UNC13A protein is increased in the cell by 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, expression of UNC13A protein is increased in the cell by between 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, expression of UNC13A protein is increased in cells 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, in which an antisense oligonucleotide is provided to a cell, 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 cell morphology, neurite length, neurotransmitter release, action potentials, neuronal synaptic plasticity, and neuronal longevity. In some embodiments, the antisense oligonucleotide consists of the sequence set forth 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 set forth in any one of SEQ ID NOs: 1059-1074, 1075-1089, and 1090-1103. In some embodiments, the antisense oligonucleotide consists of the sequence set forth 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 oligonucleotides, 2'O-methoxyethyl antisense oligonucleotides, phosphorothioate antisense oligonucleotides, and locked nucleic acid (LNA) antisense oligonucleotides.

[0144] In some embodiments, the antisense oligonucleotide comprises 21 nucleotides complementary to the sequence set forth in SEQ ID NO: 1, and the antisense oligonucleotide comprises a sequence set forth 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 set forth in SEQ ID NO: 1, wherein the antisense oligonucleotide consists of the sequence set forth 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 a cell, expression of UNC13A mRNA is increased in the cell by 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 a wild-type cell. In some embodiments, expression of UNC13A mRNA is increased in cells 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, expression of UNC13A mRNA is increased in cells 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, in which an antisense oligonucleotide is provided to a cell, expression of UNC13A cryptic exon mRNA is reduced in cells 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, in which an antisense oligonucleotide is provided to a cell, expression of UNC13A protein is increased in the cell by 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 a wild-type cell. In some embodiments, expression of UNC13A protein is increased in the cell by between about 10% and about 95%, 20% and about 85%, 30% and about 75%, 40% and about 65%, or 50% and about 55% of the level of UNC13A protein expressed in a wild-type cell. In some embodiments, expression of UNC13A protein is increased in the cell by between about 25% and about 1000%, about 50% and about 800%, about 100% and about 700%, about 200% and about 600%, or about 300% and about 500%. In some embodiments, in which an antisense oligonucleotide is provided to a cell, 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 cell morphology, neurite length, neurotransmitter release, action potentials, neuronal synaptic plasticity, and neuronal longevity. In some embodiments, the antisense oligonucleotide consists of the sequence set forth 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 set forth in any one of SEQ ID NOs: 1059-1074, 1075-1089, and 1090-1103. In some embodiments, the antisense oligonucleotide consists of the sequence set forth 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 oligonucleotides, 2'O-methoxyethyl antisense oligonucleotides, phosphorothioate antisense oligonucleotides, and locked nucleic acid (LNA) antisense oligonucleotides.

[0145] In some embodiments, the antisense oligonucleotide comprises 22 nucleotides complementary to the sequence set forth in SEQ ID NO: 1, and the antisense oligonucleotide comprises a sequence set forth 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 22 nucleotides complementary to the sequence set forth in SEQ ID NO: 1, and the antisense oligonucleotide consists of any one of the sequences set forth in 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, where an antisense oligonucleotide is provided to a cell, expression of UNC13A mRNA is increased in the cell by 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 a wild-type cell. In some embodiments, expression of UNC13A mRNA is increased in the cell by between about 10% and about 95%, 20% and about 85%, 30% and about 75%, 40% and about 65%, or 50% and about 55% of the level of UNC13A mRNA expressed in a wild-type cell. In some embodiments, expression of UNC13A mRNA is increased in the cell by between about 25% and about 1000%, about 50% and about 800%, about 100% and about 700%, about 200% and about 600%, or about 300% and about 500%.In some embodiments, where an antisense oligonucleotide is provided to a cell, expression of UNC13A cryptic exon mRNA is reduced in the cell 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, expression of UNC13A protein is increased in the cell by 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, expression of UNC13A protein is increased in the cell by between 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, expression of UNC13A protein is increased in cells 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, in which an antisense oligonucleotide is provided to a cell, 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 cell morphology, neurite length, neurotransmitter release, action potentials, neuronal synaptic plasticity, and neuronal longevity. In some embodiments, the antisense oligonucleotide consists of the sequence set forth 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 set forth in any one of SEQ ID NOs: 1059-1074, 1075-1089, and 1090-1103. In some embodiments, the antisense oligonucleotide consists of the sequence set forth 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 oligonucleotides, 2'O-methoxyethyl antisense oligonucleotides, phosphorothioate antisense oligonucleotides, and locked nucleic acid (LNA) antisense oligonucleotides.

[0146] In some embodiments, the antisense oligonucleotide comprises 23 nucleotides complementary to the sequence set forth in SEQ ID NO: 1, and the antisense oligonucleotide comprises a sequence set forth 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 23 nucleotides complementary to the sequence set forth in SEQ ID NO: 1, and the antisense oligonucleotide consists of any one of the sequences set forth in 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, where an antisense oligonucleotide is provided to a cell, expression of UNC13A mRNA is increased in the cell by 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 a wild-type cell. In some embodiments, expression of UNC13A mRNA is increased in the cell by between about 10% and about 95%, 20% and about 85%, 30% and about 75%, 40% and about 65%, or 50% and about 55% of the level of UNC13A mRNA expressed in a wild-type cell. In some embodiments, expression of UNC13A mRNA is increased in the cell by between about 25% and about 1000%, about 50% and about 800%, about 100% and about 700%, about 200% and about 600%, or about 300% and about 500%. In some embodiments in which an antisense oligonucleotide is provided to a cell, expression of UNC13A cryptic exon mRNA is reduced in the cell 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, in which an antisense oligonucleotide is provided to a cell, expression of UNC13A protein is increased in the cell by 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 a wild-type cell. In some embodiments, expression of UNC13A protein is increased in the cell by between about 10% and about 95%, 20% and about 85%, 30% and about 75%, 40% and about 65%, or 50% and about 55% of the level of UNC13A protein expressed in a wild-type cell. In some embodiments, expression of UNC13A protein is increased in the cell by between about 25% and about 1000%, about 50% and about 800%, about 100% and about 700%, about 200% and about 600%, or about 300% and about 500%. In some embodiments, in which an antisense oligonucleotide is provided to a cell, 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 cell morphology, neurite length, neurotransmitter release, action potentials, neuronal synaptic plasticity, and neuronal longevity. In some embodiments, the antisense oligonucleotide consists of the sequence set forth 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 set forth in any one of SEQ ID NOs: 1059-1074, 1075-1089, and 1090-1103. In some embodiments, the antisense oligonucleotide consists of the sequence set forth 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 oligonucleotides, 2'O-methoxyethyl antisense oligonucleotides, phosphorothioate antisense oligonucleotides, and locked nucleic acid (LNA) antisense oligonucleotides.

[0147] In some embodiments, the antisense oligonucleotide comprises 24 nucleotides complementary to the sequence set forth in SEQ ID NO: 1, and the antisense oligonucleotide comprises any of SEQ ID NOs: 53, 54, 68, 69, 83, 84, 98, 99, 113, 114, 128, 129, 143, 144, 158, 159, 173, 174, 188, 189, 203, 204, 205, 206, 207, 208, 209, 210, 211, 212, 213, 214, 215, 216, 217, 218, 219, 220, 221, 222, 223, 224, 225, 226, 227, 228, 229, 230, 231, 232, 233, 234, 235, 236, 237, 238, 239, 240, 241, 242, 243, 244, 245, 246, 247, 248, 249, 250, 251, 252, 253, 254, 255, 256, 257, 258, 259, 260, 261, 262, 263, 264, 265, 266, 267, 268, 269, 270, 271, 272, 273, 274, 275, 276, 27 18, 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 to 1115, and 1314 to 1332. In some embodiments, the antisense oligonucleotide consists of 24 nucleotides complementary to the sequence set forth in SEQ ID NO: 1, and the antisense oligonucleotide consists of any of SEQ ID NOs: 53-54, 68, 69, 83, 84, 98, 99, 113, 114, 128, 129, 143, 144, 158, 159, 173, 174, 188, 189, 203, 204, 205, 206, 207, 208, 209, 210, 211, 212, 213, 214, 215, 216, 217, 218, 219, 220, 221, 222, 223, 224, 225, 226, 227, 228, 229, 230, 231, 232, 233, 234, 235, 236, 237, 238, 239, 240, 241, 242, 243, 244, 245, 246, 247, 248, 249, 250, 251, 252, 253, 254, 255, 256, 257, 258, 259, 260, 261, 262, 263, 264, 265, 266, 267, 268, 269, 270, 271, 272, 273, 274, 275, 276, 2 18, 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, expression of UNC13A mRNA is increased in the cell by 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 a wild-type cell. In some embodiments, expression of UNC13A mRNA is increased in cells by between about 10% and about 95%, 20% and about 85%, 30% and about 75%, 40% and about 65%, or 50% and about 55% of the level of UNC13A mRNA expressed in wild-type cells.In some embodiments, expression of UNC13A mRNA is increased in the cells 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, when an antisense oligonucleotide is provided to the cells, expression of UNC13A cryptic exon mRNA is decreased in the cells 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, when an antisense oligonucleotide is provided to the cells, expression of UNC13A protein is increased in the cells by 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, expression of UNC13A protein is increased in cells by between about 10% and about 95%, 20% and about 85%, 30% and about 75%, 40% and about 65%, or 50% and about 55% of the level of UNC13A protein expressed in wild-type cells. In some embodiments, expression of UNC13A protein is increased in cells by between about 25% and about 1000%, about 50% and about 800%, about 100% and about 700%, about 200% and about 600%, or about 300% and about 500%. In some embodiments in which an antisense oligonucleotide is provided to a cell, 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 potentials, neuronal synaptic plasticity, and neuronal longevity. In some embodiments, the antisense oligonucleotide consists of the sequence set forth 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 set forth in any one of SEQ ID NOs: 1059-1074, 1075-1089, and 1090-1103.In some embodiments, the antisense oligonucleotide consists of 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 oligonucleotides, 2'O-methoxyethyl antisense oligonucleotides, phosphorothioate antisense oligonucleotides, and locked nucleic acid (LNA) antisense oligonucleotides.

[0148] In some embodiments, the antisense oligonucleotide comprises 25 nucleotides complementary to the sequence set forth in SEQ ID NO: 1, and the antisense oligonucleotide comprises a sequence set forth in any one of SEQ ID NOs: 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 set forth in SEQ ID NO: 1, and the antisense oligonucleotide consists of any one of the sequences set forth in SEQ ID NOs: 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, where an antisense oligonucleotide is provided to a cell, expression of UNC13A mRNA is increased in the cell by 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 a wild-type cell. In some embodiments, expression of UNC13A mRNA is increased in the cell by between about 10% and about 95%, 20% and about 85%, 30% and about 75%, 40% and about 65%, or 50% and about 55% of the level of UNC13A mRNA expressed in a wild-type cell. In some embodiments, expression of UNC13A mRNA is increased in the cell by between about 25% and about 1000%, about 50% and about 800%, about 100% and about 700%, about 200% and about 600%, or about 300% and about 500%.In some embodiments, where an antisense oligonucleotide is provided to a cell, expression of UNC13A cryptic exon mRNA is reduced in the cell 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, expression of UNC13A protein is increased in the cell by 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, expression of UNC13A protein is increased in the cell by between 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, expression of UNC13A protein is increased in cells 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, in which an antisense oligonucleotide is provided to a cell, 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 cell morphology, neurite length, neurotransmitter release, action potentials, neuronal synaptic plasticity, and neuronal longevity. In some embodiments, the antisense oligonucleotide consists of the sequence set forth 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 set forth in any one of SEQ ID NOs: 1059-1074, 1075-1089, and 1090-1103. In some embodiments, the antisense oligonucleotide consists of the sequence set forth 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 oligonucleotides, 2'O-methoxyethyl antisense oligonucleotides, phosphorothioate antisense oligonucleotides, and locked nucleic acid (LNA) antisense oligonucleotides.

[0149] 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 oligonucleotides, 2'O-methoxyethyl antisense oligonucleotides, phosphorothioate antisense oligonucleotides, and LNA antisense oligonucleotides. In some embodiments, the modification is a 2'OMe antisense oligonucleotide.

[0150] 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' and 3' ends of the antisense oligonucleotide comprise a modification, and at least one, at least two, at least three, at least four, or at least five nucleotides are modified at each of the 5' and 3' ends of the antisense oligonucleotide. In some embodiments, the modification is selected from the group consisting of 2'OMe antisense oligonucleotides, 2'O-methoxyethyl antisense oligonucleotides, phosphorothioate antisense oligonucleotides, and LNA antisense oligonucleotides. In some embodiments, the modification is a 2'OMe antisense oligonucleotide.

[0151] In some embodiments, the antisense oligonucleotide comprises a modification, 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 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 oligonucleotides, 2'O-methoxyethyl antisense oligonucleotides, phosphorothioate antisense oligonucleotides, and LNA antisense oligonucleotides. In some embodiments, the modification is a 2'OMe antisense oligonucleotide.

[0152] In some embodiments, the antisense oligonucleotide contains one or more modified nucleotides, such as nucleotides other than ribonucleotides (2'-hydroxyl nucleotides), including, but not limited to, deoxynucleotides, nucleotide mimics, abasic nucleotides, 2'-modified nucleotides, 3'-3'-linked (inverted) nucleotides, unnatural base-containing nucleotides, bridged nucleotides, peptide nucleic acids, 2',3'-seconucleotide mimics, locked nucleotides, 3'-O-methoxy nucleotides, 2'-F-arabinonucleotides, morpholino nucleotides, vinylphosphonate deoxyribonucleotides, and vinylphosphonate nucleotides, 2'-O-methyl nucleotides, 2'-deoxy-2'-fluoronucleotides, 2'-deoxynucleotides, 2'-methoxyethyl nucleotides, 2'-amino nucleotides, and 2'-alkyl nucleotides.

[0153] In some embodiments, the antisense oligonucleotide is a modified antisense oligonucleotide. In some embodiments, the modified antisense oligonucleotide includes a phosphoramidate morpholino oligonucleotide, a phosphorodiamidate morpholino oligonucleotide, a phosphorothioate modified oligonucleotide, a 2'O-methyl (2'O-Me) modified oligonucleotide, a peptide nucleic acid (PNA), a locked nucleic acid (LNA), a phosphorodithioate oligonucleotide, a 2'O-methoxyethyl (2'-MOE) modified oligonucleotide, a 2'-fluoro modified oligonucleotide, a 2'O,4'C-ethylene bridged nucleic acid (ENA), a tricyclo-DNA, a tricyclo-DNA phosphorothioate nucleotide, a constrained ethyl bridged nucleotide, a 2'-O-[2-(N-methylcarbamoyl)ethyl] modified oligonucleotide, a morpholino oligonucleotide, and a peptide-conjugated phosphoramidate morpholino oligonucleotide (PPMO), or any combination thereof. In some embodiments, the modification of the antisense oligonucleotide increases the stability of the oligonucleotide, the effectiveness of reducing cryptic exon inclusion, or the efficiency of reducing cryptic exon inclusion. In some embodiments, modifications of the antisense oligonucleotide improve delivery of the oligonucleotide to the UNC13 mRNA molecule.

[0154] In some embodiments, the ASO comprises a modification that enhances the pharmacokinetic or biodistribution properties of the ASO trigger or conjugate to which it is attached, improving cell- or tissue-specific distribution and cell-specific uptake of the conjugate. In some cases, binding of the group to a cell or cell receptor can initiate endocytosis. The modification can be monovalent, bivalent, trivalent, tetravalent, or higher. Exemplary targeting groups include, but are not limited to, compounds with affinity for cell surface molecules, cell receptor ligands, haptens, antibodies, monoclonal antibodies, antibody fragments, and antibody mimetics with affinity for cell surface molecules.

[0155] In some embodiments, the modification comprises modifying ribonucleic acid (RNA) to deoxyribonucleic acid (DNA), threose nucleic acid (TNA), glycol nucleic acid (GNA), peptide nucleic acid (PNA), locked nucleic acid (LNA), or hybrids thereof.

[0156] In some embodiments, the ASO comprises a modified sugar moiety. In some embodiments, the ASO comprises a sugar surrogate. In certain such embodiments, the oxygen atom of the sugar moiety is replaced with, for example, a sulfur, carbon, or nitrogen atom. In certain such embodiments, such modified sugar moieties also comprise bridging and / or non-bridging substituents. In some embodiments, morpholinos are modified by adding or altering various substituents from the morpholino structure described above.

[0157] In some embodiments, ASOs contain modified variants of nucleosides that maintain proper base pairing. In some embodiments, ASOs contain modified A, C, G, or U. In one embodiment, a modified ASO contains modified C, e.g., 5-methylcytosine or 5-hydroxymethylcytosine, modified U, e.g., 5-methyluridine, or a substitution with thymine, or modified A, e.g., Ne-methyladenine. In one embodiment, an ASO may contain a mixture of unmodified and modified nucleosides.

[0158] In some embodiments, the ASO comprises modified internucleotidic linkages. 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 from mRNA (alternating phosphate and ribose), LNA (locked nucleic acid), tcDNA (tricyclic DNA), cEt (constrained ethyl-bridged nucleic acid), ENA (ethylene-bridged nucleic acid), HNA (hexitol nucleic acid), TNA (threose nucleic acid), PMO (phosphorodiamidate morpholino oligomer), PNA (peptide nucleic acid), 2-OMe-RNA, 2'-O-methoxyethyl (MOE) nucleic acid, or 2-O-(2-methylcarbomoyl (MCE) nucleotides, or any combination thereof.

[0159] In some embodiments, the antisense oligonucleotide is used in a method of treatment or in the manufacture of a medicament for use in treatment, and the antisense oligonucleotide is administered to a patient in need of increased UNC13A mRNA expression. In some embodiments, UNC13A mRNA expression is increased 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, UNC13A mRNA expression is increased in cells 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, UNC13A mRNA expression is increased in cells by about 100%, about 200%, about 300%, about 400%, or about 500%. In some embodiments, the antisense oligonucleotide increases UNC13A protein expression in a cell by 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 a wild-type cell. In some embodiments, UNC13A mRNA expression is increased in a cell by between about 10% and about 95%, 20% and about 85%, 30% and about 75%, 40% and about 65%, or 50% and about 55% of the level of UNC13A mRNA expressed in a wild-type cell. In some embodiments, the antisense oligonucleotide comprises or consists of a sequence set forth 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 set forth 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 set forth in any one of SEQ ID NOs: 1059-1074, 1075-1089, and 1090-1103.In some embodiments, the antisense oligonucleotide consists of the sequence set forth in any one of SEQ ID NOs: 1248-1270, 1271-1292, and 1293-1313.

[0160] In some embodiments, the antisense oligonucleotide restores UNC13A mRNA expression in a 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 a wild-type cell. In some embodiments, the antisense oligonucleotide restores UNC13A protein expression in a cell to between about 10% and about 95%, 20% and about 85%, 30% and about 75%, 40% and about 65%, or 50% and about 55% of the level of UNC13A protein expressed in a wild-type cell. In some embodiments, the antisense oligonucleotide comprises or consists of a sequence set forth 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 set forth 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 set forth in any one of SEQ ID NOs: 1059-1074, 1075-1089, and 1090-1103. In some embodiments, the antisense oligonucleotide consists of the sequence set forth in any one of SEQ ID NOs: 1248-1270, 1271-1292, and 1293-1313.

[0161] In some embodiments, the antisense oligonucleotide is used in a method of treatment or in the manufacture of a medicament for use in treatment, wherein the antisense oligonucleotide is administered to a patient in need of reduced expression of UNC13A cryptic exon mRNA. In some embodiments, expression of UNC13A cryptic exon mRNA is reduced in cells by at least 10%, at least 20%, at least 30%, at least 40%, or at least 50%. In some embodiments, expression of UNC13A cryptic exon mRNA is reduced in cells 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, expression of UNC13A cryptic exon mRNA is reduced in cells by about 10%, about 20%, about 30%, about 40%, or about 50%. In some embodiments, the antisense oligonucleotide comprises or consists of a sequence set forth 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 set forth 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 set forth in any one of SEQ ID NOs: 1059-1074, 1075-1089, and 1090-1103. In some embodiments, the antisense oligonucleotide consists of the sequence set forth in any one of SEQ ID NOs: 1248-1270, 1271-1292, and 1293-1313.

[0162] In some embodiments, the antisense oligonucleotide reduces 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 an shRNA targeting TARDBP. In some embodiments, the antisense oligonucleotide comprises or consists of a sequence set forth in any one of SEQ ID NOs: 20-355, 400-638, 810-959, or 961-1332. In some embodiments, the antisense oligonucleotide consists of a sequence set forth 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 set forth in any one of SEQ ID NOs: 1059 to 1074, 1075 to 1089, and 1090 to 1103. In some embodiments, the antisense oligonucleotide consists of the sequence set forth in any one of SEQ ID NOs: 1248 to 1270, 1271 to 1292, and 1293 to 1313.

[0163] In some embodiments, the antisense oligonucleotide is used in a method of treatment or in the manufacture of a medicament for use in treatment, and the antisense oligonucleotide is administered to a patient in need of increased expression of UNC13A protein. In some embodiments, UNC13A protein expression is increased 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, UNC13A protein expression is increased in cells 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, UNC13A protein expression is increased in cells by about 100%, about 200%, about 300%, about 400%, or about 500%. In some embodiments, the antisense oligonucleotide comprises or consists of the sequence set forth 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 set forth 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 set forth in any one of SEQ ID NOs: 1059-1074, 1075-1089, and 1090-1103. In some embodiments, the antisense oligonucleotide consists of the sequence set forth in any one of SEQ ID NOs: 1248-1270, 1271-1292, and 1293-1313.

[0164] In some embodiments, the antisense oligonucleotide is used in a method of treatment or in the manufacture of a medicament for use in treatment, wherein the antisense oligonucleotide is administered to a patient in need of increasing UNC13A protein expression to a level sufficient to improve one or more neurological functions. In some embodiments, the one or more neurological functions are selected from the group consisting of neuronal signaling, neuronal morphology, neurite length, neurotransmitter release, action potentials, neuronal synaptic plasticity, and neuronal longevity. In some embodiments, the antisense oligonucleotide improves one, two, three, four, five, or six neuronal functions selected from the group consisting of neuronal signaling, neuronal morphology, neurite length, neurotransmitter release, action potentials, neuronal synaptic plasticity, and neuronal longevity. In some embodiments, the antisense oligonucleotide improves at least one, at least two, at least three, at least four, or at least five neuronal functions selected from the group consisting of neuronal signaling, neuronal morphology, neurite length, neurotransmitter release, action potentials, neuronal synaptic plasticity, and neuronal longevity. In some embodiments, the antisense oligonucleotide comprises or consists of the sequence set forth 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 set forth 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 set forth in any one of SEQ ID NOs: 1059-1074, 1075-1089, and 1090-1103. In some embodiments, the antisense oligonucleotide consists of the sequence set forth in any one of SEQ ID NOs: 1248-1270, 1271-1292, and 1293-1313.

[0165] In some embodiments, the antisense oligonucleotide restores expression of UNC13A protein in a 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 a wild-type cell. In some embodiments, the antisense oligonucleotide comprises or consists of a sequence set forth in any one of SEQ ID NOs: 20-355, 400-638, 810-959, or 961-1332. In some embodiments, the antisense oligonucleotide consists of a sequence set forth 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 set forth in any one of SEQ ID NOs: 1059-1074, 1075-1089, and 1090-1103. In some embodiments, the antisense oligonucleotide consists of the sequence set forth in any one of SEQ ID NOs: 1248-1270, 1271-1292, and 1293-1313.

[0166] In some embodiments, the antisense oligonucleotide increases UNC13A mRNA expression about 4 days, about 8 days, about 12 days, about 16 days, about 20 days, or about 24 days after contacting the cells with the ASO. In some embodiments, the antisense oligonucleotide is provided to the cells in multiple doses. In some embodiments, the antisense oligonucleotide increases UNC13A mRNA expression about 4 days, about 8 days, about 12 days, about 16 days, about 20 days, or about 24 days after administration of the ASO. In some embodiments, the antisense oligonucleotide increases UNC13A mRNA expression such that UNC13A expression is fully restored after administration of the ASO.

[0167] In some embodiments, the antisense oligonucleotide reduces UNC13A cryptic exon mRNA expression about 4 days, about 8 days, about 12 days, about 16 days, about 20 days, or about 24 days after contacting the cells with the ASO. In some embodiments, the antisense oligonucleotide is provided to the cells in multiple doses. In some embodiments, the antisense oligonucleotide reduces UNC13A cryptic exon mRNA expression about 4 days, about 8 days, about 12 days, about 16 days, about 20 days, or about 24 days after administration of the ASO. In some embodiments, the antisense oligonucleotide reduces UNC13A cryptic exon mRNA expression such that expression of UNC13A cryptic exon mRNA is completely reduced after administration of the ASO.

[0168] In some embodiments, the antisense oligonucleotide increases UNC13A protein expression about 4 days, about 8 days, about 12 days, about 16 days, about 20 days, or about 24 days after contacting the cells with the ASO. In some embodiments, the antisense oligonucleotide is provided to the cells in multiple doses. In some embodiments, the antisense oligonucleotide increases UNC13A protein expression about 4 days, about 8 days, about 12 days, about 16 days, about 20 days, or about 24 days after administration of the ASO. In some embodiments, the antisense oligonucleotide increases UNC13A protein expression such that the increase in UNC13A protein expression is completely restored after administration of the ASO.

[0169] In some embodiments, the antisense oligonucleotide is provided to a cell, and the cell is a human cell or a mouse cell. In some embodiments, the cell is a human cell. In some embodiments, the cell is a mouse cell. In some embodiments, the antisense oligonucleotide is provided to a cell, and the cell is a neuron. In some embodiments, the cell is a human neuron. In some embodiments, the cell is a mouse 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 the 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 repeats→3-5 CATC repeats), and rs62121687 (hg38 chr19:17,642,351 C→A), or any combination thereof. In some embodiments, the cell comprises a mutation associated with amyotrophic lateral sclerosis (ALS) or frontotemporal dementia (FTD). In some embodiments, the antisense oligonucleotide comprises or consists of a sequence set forth in any one of SEQ ID NOs: 20-355, 400-638, 810-959, or 961-1332.

[0170] In some embodiments, the antisense oligonucleotide is provided to a cell, and 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, and the cell is heterozygous or homozygous for the 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 repeats→3-5 CATC repeats), and rs62121687 (hg38 chr19:17,642,351 C→A), or any combination thereof. In some embodiments, the risk allele is rs12973192 (hg38 chr19:17,642,430 C→G). In some embodiments, the antisense oligonucleotide is provided to a cell containing a SNP, wherein the SNP is rs12973192 (hg38 chr19:17,642,430 C→G). In some embodiments, the cell is selected from the group consisting of GM25256 (GM or GM strain), NDS00262 (005 or 005 strain), NDS00209 (4048 or 4048 strain), and NDS00235 (4157 or 4157 strain). In some embodiments, the cell is a healthy cell selected from the group consisting of GM25256 (GM or GM strain) and NDS00262 (005 or 005 strain).In some embodiments, the cell is a cell comprising a mutation in the TDP-43 protein selected from the group consisting of NDS00209 (strain 4048 or 4048) and NDS00235 (strain 4157 or 4157). In some embodiments, the cell is NDS00209 (strain 4048 or 4048) and the mutation in the TDP-43 protein is G298S. In some embodiments, the cell is NDS00235 (strain 4157 or 4157) and the mutation in the TDP-43 protein is A382T. In some embodiments, the antisense oligonucleotide comprises or consists of a sequence set forth in any one of SEQ ID NOs: 20-355, 400-638, 810-959, or 961-1332.

[0171] In some embodiments, the antisense oligonucleotide is active in one or more iPSC-neurons. In some embodiments, the antisense oligonucleotide is active in a plurality of iPSC-neurons with different genetic backgrounds, disease mutations, or UNC13A SNP status. In some embodiments, the antisense oligonucleotide is active in a plurality of iPSC-neurons with different genetic backgrounds, disease mutations, and UNC13A SNP status. In some embodiments, the antisense oligonucleotide comprises or consists of a sequence set forth in any one of SEQ ID NOs: 20-355, 400-638, 810-959, or 961-1332.

[0172] In some embodiments, an antisense oligonucleotide is provided to a cell, and the cell expresses low levels 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% of TDP-43 protein. In some embodiments, the antisense oligonucleotide comprises or consists of a sequence set forth in any one of SEQ ID NOs: 20-355, 400-638, 810-959, or 961-1332.

[0173] In some embodiments, the antisense oligonucleotides are provided to the cells by transfection, hi some embodiments, the antisense oligonucleotides are provided to the cells by other delivery methods known in the art.

[0174] In some embodiments, the antisense oligonucleotide does not cause any changes in cell morphology. In some embodiments, the antisense oligonucleotide does not cause neurotoxicity. In some embodiments, the antisense oligonucleotide does not bind to genomic sequences other than the sequence set forth 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 a sequence set forth in any one of SEQ ID NOs: 20-355, 400-638, 810-959, or 961-1332.

[0175] In some embodiments, the antisense oligonucleotides described herein reduce expression of UNC13A cryptic exon mRNA. In some cases, this reduction in UNC13A cryptic exon mRNA alone does not indicate upregulation of full-length UNC13A mRNA. In some cases, the antisense oligonucleotides prevent proper splicing of the intron.

[0176] In some embodiments, the antisense oligonucleotide occupies an overlapping region with other antisense oligonucleotides described herein. In some cases, two antisense oligonucleotides targeting adjacent and / or overlapping regions of UNC13A mRNA have different activities, with the first antisense oligonucleotide being active and the second antisense oligonucleotide being inactive. In some cases, the two antisense oligonucleotides targeting adjacent and / or overlapping regions of UNC13A mRNA are the same nucleotide length or different nucleotide lengths. In some cases, a single nucleotide shift in the targeting sequence affects the ability of the antisense oligonucleotide to block the splicing mechanism. In some cases, a single nucleotide shift in the targeting sequence results in significantly different antisense oligonucleotide activities, with the first antisense oligonucleotide being active and the second antisense oligonucleotide being inactive.

[0177] In some cases, antisense oligonucleotides described herein consisting of at least 18 nucleotides are more active. In some cases, antisense oligonucleotides consisting of at least 20 nucleotides are most potent, particularly antisense oligonucleotides consisting of 20, 21, or 22 nucleotides. In some cases, selected antisense oligonucleotides consisting of 13-17 nucleotides, particularly 13 or 15 nucleotides, are active. In some embodiments, this represents the minimum region that needs to be blocked to have the desired effect. This emphasizes the importance of both the positioning and length of the antisense oligonucleotide on the UNC13A mRNA for optimal performance.

[0178] In some cases, the antisense oligonucleotide is provided to the cells for a long period of time. In some cases, the antisense oligonucleotide restores UNC13A levels to those of wild-type cells. In some cases, the cells are neurons from one or more different iPSC lines, such as GM25256 (GM or GM line) and NDS00262 (005 or 005 line).

[0179] III. Pharmaceutical Compositions In some aspects, the present disclosure provides a pharmaceutical composition comprising an UNC13A cryptic exon splice variant-specific inhibitor described herein and a pharmaceutically acceptable carrier. As used herein, the term "pharmaceutically acceptable" refers to compounds, materials, compositions, and / or dosage forms that, within the scope of sound medical judgment, are suitable for use in contact with cells and / or tissues without excessive toxicity, irritation, allergic response, or other problem or complication commensurate with a reasonable benefit / risk ratio.

[0180] As used herein, the term "pharmaceutically acceptable carrier" refers to a pharmaceutically acceptable material, composition, or carrier, such as a liquid or solid filler, stabilizer, dispersant, suspending agent, diluent, excipient, thickener, solvent, or encapsulating material, that is involved in carrying or transporting a compound useful within the present invention within or to a patient so that it may perform its intended function. Each carrier must be "acceptable" in the sense of being compatible with the other ingredients of the formulation and not deleterious to the cells or tissues with which it is contacted. Additional ingredients that may be included in pharmaceutical compositions used to practice the present invention are known in the art and are described, for example, in Remington's Pharmaceutical Sciences (Genaro, Ed., Mack Publishing Co., 1985, Easton, PA), incorporated herein by reference.

[0181] As is well known in the medical arts, the dosage for any one patient will depend on many factors, including the patient's size, weight, body surface area, age, the level of UNC13A cryptic exon splice variant-specific inhibitor needed to achieve therapeutic effect, the stability of the UNC13A cryptic exon splice variant-specific inhibitor, the particular disease being treated, the stage of the disease, sex, the time and route of administration, general health, and other drugs being administered concomitantly.

[0182] Pharmaceutical compositions may be administered in a manner appropriate for the disease or condition being treated (or prevented), as determined by one of ordinary skill in the art. The appropriate dose and suitable duration and frequency of administration of the composition will depend on factors such as the patient's health, the patient's size (i.e., body weight, mass, or volume), the type and severity of the patient's disease, the particular form of the active ingredient, and the method of administration. Generally, an appropriate dose and treatment regimen will 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 remissions, or longer disease-free and / or overall survival, or reduced severity of symptoms). For prophylactic use, the dose should be sufficient to prevent, delay the onset of, or reduce the severity of disease associated with the disease or disorder. The prophylactic benefit of the compositions administered in accordance with 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 using appropriate statistical, biological, and clinical methods and techniques, all of which can be readily performed by one of ordinary skill in the art.

[0183] The composition (e.g., pharmaceutical composition) may be administered by any route, including enteral (e.g., oral), parenteral, intravenous, intramuscular, intraarterial, intramedullary, intrathecal, subpial, intraparenchymal, intrastriatal, intracranial, intracisternal, intracerebral, intraventricular, intraocular, intraventricular, intralumbar, subcutaneous, transdermal, intradermal, rectal, vaginal, intraperitoneal, topical (by powder, ointment, cream, and / or drops), mucosal, intranasal, buccal, sublingual, intratracheal instillation, intratracheal instillation, and / or by inhalation, and / or oral spray, nasal spray, and / or aerosol. Generally, the most appropriate administration route 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 injected directly into the subject's CNS. In some embodiments, the direct injection into the CNS is an intracerebral injection, an intraparenchymal injection, an intrathecal injection, an intrathecal injection, or any combination thereof. In some embodiments, the direct injection into the CNS is a direct injection into the cerebrospinal fluid (CSF) of a subject, and optionally, the direct injection is an intracisternal injection, an intraventricular injection, and / or an intralumbar injection.

[0184] IV. Methods of Use for UNC13A Cryptic Splice Variant Inhibitors The antisense oligonucleotides provided herein have several beneficial properties that make them useful for increasing UNC13A protein expression, reducing UNC13A cryptic exon inclusion, and treating neurodegenerative diseases, as further described below.

[0185] A. Methods for increasing UNC13A protein expression using ASOs Provided herein are methods for increasing expression of UNC13A protein in cells, comprising contacting the cells with an antisense oligonucleotide provided herein. In some embodiments, expression of UNC13A protein is increased by at least 10% after contacting the cells with the antisense oligonucleotide. In some embodiments, 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 levels before contact with the antisense oligonucleotide. In some embodiments, expression of UNC13A is increased by at least 1.1, at least 1.2, at least 1.3, at least 1.5, at least 1.5, or at least 2-fold compared to levels 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 sequence set forth in SEQ ID NOs: 20-355, or SEQ ID NOs: 20-40, or SEQ ID NOs: 41-355, or SEQ ID NOs: 810-957. In some embodiments, the antisense oligonucleotide comprises or consists of the sequence set forth 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 the sequence set forth 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 comprises or consists of the sequence set forth in any one of SEQ ID NOs: 1059-1074, 1075-1089, and 1090-1103.In some embodiments, the antisense oligonucleotide comprises or consists of a sequence set forth in any one of SEQ ID NOs: 1248-1270, 1271-1292, and 1293-1313. In some embodiments, the antisense oligonucleotide comprises a modification.

[0186] In some embodiments, UNC13A protein expression is increased to normal levels after contacting cells with the antisense oligonucleotide. In some embodiments, UNC13A protein expression is increased to the level of an individual without a mutation in TDP-43. In some embodiments, UNC13A protein expression is increased to the level of an individual without reduced TDP-43 protein expression. In some embodiments, UNC13A protein expression is increased to the level of an individual without a risk allele associated with ALS or FTD. In some embodiments, UNC13A expression is increased to the level of an individual without ALS or FTD. In some embodiments, the antisense oligonucleotide comprises or consists of a sequence set forth in SEQ ID NOs: 20-355, 20-40, 41-355, or 810-957. In some embodiments, the antisense oligonucleotide comprises or consists of a sequence set forth 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 the sequence set forth 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 comprises or consists of the sequence set forth in any one of SEQ ID NOs: 1059-1074, 1075-1089, and 1090-1103. In some embodiments, the antisense oligonucleotide comprises or consists of the sequence set forth in any one of SEQ ID NOs: 1248-1270, 1271-1292, and 1293-1313. In some embodiments, the antisense oligonucleotide comprises a modification.

[0187] In some embodiments, the cells have a low level of UNC13A protein expression before contacting the cells with the antisense oligonucleotide. In some embodiments, the cells have a mutation in TDP-43 or reduced expression of TDP-43 protein. In some embodiments, the cells have a level of UNC13A protein expression associated with a TDP-43 proteinopathy. In some embodiments, the cells have a level of UNC13A protein associated with a risk allele associated with ALS or FTD. In some embodiments, the cells have a level of UNC13A associated with ALS or FTD. In some embodiments, the antisense oligonucleotide comprises or consists of a sequence set forth in SEQ ID NOs: 20-355, or 20-40, or 41-355, or 810-957. In some embodiments, the antisense oligonucleotide comprises or consists of a sequence set forth in any one of SEQ ID NOs: 20-355, 400-638, 810-959, or 961-1332. In some embodiments, the antisense oligonucleotide comprises a modification.

[0188] In some embodiments, the cells have a low level of UNC13A protein expression compared to normal or wild-type cells prior to contacting the cells with the antisense oligonucleotide. In some embodiments, normal cells are cells that do not have a mutation in TDP-43, have normal expression levels of TDP-43 protein, do not have a risk allele associated with ALS or FTD, and / or are not obtained from an individual with ALS or FTD. In some embodiments, the cells have undetectable levels of UNC13A protein expression prior to contacting with the ASO. In some embodiments, the cells express less than 80%, less than 70%, less than 60%, or less than 50% of the normal level of UNC13A protein. In some embodiments, the antisense oligonucleotide comprises or consists of a sequence set forth in SEQ ID NOs: 20-355, or SEQ ID NOs: 20-40, or SEQ ID NOs: 41-355, or SEQ ID NOs: 810-957. In some embodiments, the antisense oligonucleotide comprises or consists of a sequence set forth in any one of SEQ ID NOs: 20 to 355, 400 to 638, 810 to 959, or 961 to 1332. In some embodiments, the antisense oligonucleotide comprises a modification.

[0189] In some aspects, provided herein are methods for increasing expression of UNC13A protein in a cell by at least 10%, comprising contacting a cell with an antisense oligonucleotide comprising 13 to 30 nucleotides, wherein the antisense oligonucleotide is complementary to the sequence set forth in SEQ ID NO: 1. In some aspects, provided herein are methods for increasing expression of UNC13A protein in a cell by at least 10%, comprising contacting a cell with an antisense oligonucleotide comprising 13 to 30 nucleotides, wherein the antisense oligonucleotide is complementary to the sequence set forth in any one of SEQ ID NOs: 1339 to 1342, or a portion thereof. In some embodiments, the antisense oligonucleotide that modulates UNC13A has a sequence complementary to a sequence in 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.

[0190] In some embodiments, the antisense oligonucleotide is complementary to the sequence set forth 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 to the sequence set forth in SEQ ID NO: 1.

[0191] In some embodiments, the antisense oligonucleotide is complementary to the sequence set forth in any one of SEQ ID NOs: 700-703. In some embodiments, the antisense oligonucleotide is complementary to the sequence set forth 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 to the sequence set forth in SEQ ID NO: 701 or 703. In some embodiments, the antisense oligonucleotide is fully complementary to the sequence set forth in SEQ ID NO: 701 or 703. In some embodiments, the antisense oligonucleotide is complementary to the sequence set forth 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 to the sequence set forth in SEQ ID NO: 700. In some embodiments, the antisense oligonucleotide is fully complementary to the sequence set forth in SEQ ID NO: 700. In some embodiments, the antisense oligonucleotide is complementary to the sequence set forth 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 to the sequence set forth in SEQ ID NO: 701. In some embodiments, the antisense oligonucleotide is fully complementary to the sequence set forth in SEQ ID NO: 701. In some embodiments, the antisense oligonucleotide is complementary to the sequence set forth 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 to the sequence set forth in SEQ ID NO: 702. In some embodiments, the antisense oligonucleotide is fully complementary to the sequence set forth in SEQ ID NO: 702.In some embodiments, the antisense oligonucleotide is complementary to the sequence set forth 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 to the sequence set forth in SEQ ID NO: 703. In some embodiments, the antisense oligonucleotide is fully complementary to the sequence set forth in SEQ ID NO: 703.

[0192] 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 set forth 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 set forth 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 set forth in SEQ ID NO: 703.

[0193] In some cases, the methods provided herein include contacting a cell with an antisense oligonucleotide, wherein the antisense oligonucleotide is complementary to a 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, the UNC13A sequence being uniquely sensitive to antisense oligonucleotide binding. In some cases, regions of UNC13A mRNA are identified as being more sensitive than other regions. These regions are determined herein to be sensitive to a number of factors, including, but not limited to, antisense oligonucleotide length, composition, and location on the UNC13A mRNA. Minimal regions of UNC13A mRNA are specifically targeted to maximize antisense oligonucleotide activity in modulating UNC13A mRNA splicing. In some embodiments, these highly responsive minimal regions of UNC13A mRNA are referred to as hotspots.

[0194] In some embodiments, a minimal region 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 is identified that is 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 inclusion of the UNC13A cryptic exon. In some embodiments, the minimal region is 3-30, 4-26, 5-22, 6-18, 7-14, or 8-10 nucleotides in length. In some embodiments, the minimal region is 30 nucleotides or less, 25 nucleotides or less, 20 nucleotides or less, or 15 nucleotides or less 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 set forth in any one of SEQ ID NOs: 1339-1342, or a portion thereof. In some embodiments, the minimal region comprises the sequence set forth in SEQ ID NO: 1339, or a portion thereof. In some embodiments, the minimal region comprises the sequence set forth in SEQ ID NO: 1340, or a portion thereof. In some embodiments, the minimal region comprises the sequence set forth in SEQ ID NO: 1341, or a portion thereof. In some embodiments, the minimal region comprises the sequence set forth in SEQ ID NO: 1342, or a portion thereof. In some embodiments, combining the subregion, or a portion thereof, with the antisense oligonucleotide results in improved splice regulatory activity. In some embodiments, combining the minimal region or a portion thereof with an antisense oligonucleotide results in increased UNC13A mRNA expression, hi some embodiments, combining the minimal region or a portion thereof with an antisense oligonucleotide results in decreased UNC13A cryptic exon mRNA expression.In some embodiments, binding of the minimal region or a portion thereof with an antisense oligonucleotide results in an increase in UNC13A protein expression.

[0195] In some instances, the methods provided herein include contacting a cell with an antisense oligonucleotide comprising 13 to 30 nucleotides, wherein the antisense oligonucleotide is complementary to the sequence set forth in any one of SEQ ID NOs: 1339-1342, or a portion thereof. In some embodiments, the antisense oligonucleotide is complementary to the sequence set forth in SEQ ID NO: 1339, or a portion thereof. In some embodiments, the antisense oligonucleotide is complementary to the sequence set forth in SEQ ID NO: 1340, or a portion thereof. In some embodiments, the antisense oligonucleotide is complementary to the sequence set forth in SEQ ID NO: 1341, or a portion thereof. In some embodiments, the antisense oligonucleotide is complementary to the sequence set forth 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 set forth 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 set forth 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 consecutive nucleotides of the sequence set forth 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 set forth 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 set forth 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 set forth 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 set forth 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 set forth in SEQ ID NO: 1342.

[0196] 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 any one of SEQ ID NOs: 1339-1342, or a portion thereof, results in increased UNC13A splice regulatory activity. In some embodiments, binding of the antisense oligonucleotide to any one of SEQ ID NOs: 1339-1342, or a portion thereof, results in increased expression of UNC13A mRNA. In some embodiments, expression of UNC13A mRNA is increased in the cell by 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, expression of UNC13A mRNA is increased in the cell by between about 10% and about 95%, 20% and about 85%, 30% and about 75%, 40% and about 65%, or 50% and about 55% of the level of UNC13A mRNA expressed in wild-type cells. In some embodiments, expression of UNC13A mRNA is increased 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, expression of UNC13A mRNA is increased in cells 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 an antisense oligonucleotide to a sequence or portion set forth in any one of SEQ ID NOs: 1339-1342 results in a decrease in UNC13A cryptic exon mRNA expression. In some embodiments, expression of UNC13A cryptic exon mRNA is decreased in cells by at least 10%, at least 20%, at least 30%, at least 40%, or at least 50%. In some embodiments, expression of UNC13A cryptic exon mRNA is reduced in cells by about 10% to about 90%, about 20% to about 80%, about 30% to about 70%, or about 40% to about 60%.

[0197] In some embodiments, the wild-type cell is from an individual who does not have amyotrophic lateral sclerosis (ALS) or frontotemporal dementia (FTD). In some embodiments, the wild-type cell is a cell that expresses normal levels of TDP-43 protein. In some embodiments, the wild-type cell is a cell that does not produce a transcript of UNC13A that contains a cryptic exon.

[0198] In some aspects, provided herein are methods for increasing expression of UNC13A protein in a cell by at least 10%, comprising contacting the cell with an antisense oligonucleotide, wherein the antisense oligonucleotide comprises a sequence set forth 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 13-30 nucleotides. In some embodiments, the antisense oligonucleotide comprises or consists of 13-30, 15-28, 17-26, 19-24, or 21-23 nucleotides. In some embodiments, the antisense oligonucleotide comprises or consists of 13-25 nucleotides. In some embodiments, the antisense oligonucleotide comprises 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 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, the antisense oligonucleotide comprises or consists of 25 nucleotides. In some embodiments, the antisense oligonucleotide is provided to a cell, and expression of UNC13A mRNA is increased in the cell by 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, expression of UNC13A mRNA is increased in the cell by between about 10% and about 95%, 20% and about 85%, 30% and about 75%, 40% and about 65%, or 50% and about 55% of the level of UNC13A mRNA expressed in wild-type cells. In some embodiments, expression of UNC13A mRNA is increased in the cells 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, when an antisense oligonucleotide is provided to the cells, expression of UNC13A cryptic exon mRNA is decreased in the cells 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, when an antisense oligonucleotide is provided to the cells, expression of UNC13A protein is increased in the cells by 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, expression of UNC13A protein is increased in cells by between about 10% and about 95%, 20% and about 85%, 30% and about 75%, 40% and about 65%, or 50% and about 55% of the level of UNC13A protein expressed in wild-type cells. In some embodiments, expression of UNC13A protein is increased in cells by between about 25% and about 1000%, about 50% and about 800%, about 100% and about 700%, about 200% and about 600%, or about 300% and about 500%. In some embodiments in which an antisense oligonucleotide is provided to a cell, 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 potentials, neuronal synaptic plasticity, and neuronal longevity. In some embodiments, the antisense oligonucleotide consists of the sequence set forth 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 set forth in any one of SEQ ID NOs: 1059-1074, 1075-1089, and 1090-1103. In some embodiments, the antisense oligonucleotide consists of the sequence set forth 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 oligonucleotides, 2'O-methoxyethyl antisense oligonucleotides, phosphorothioate antisense oligonucleotides, and LNA antisense oligonucleotides.

[0199] In some embodiments, a method for increasing expression of UNC13A protein in a cell by at least 10% comprises contacting a cell with an antisense oligonucleotide, wherein the antisense oligonucleotide is complementary to the sequence set forth in SEQ ID NO: 1. In some embodiments, the antisense oligonucleotide comprises or consists of the sequence set forth 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 13-30 nucleotides. In some embodiments, the antisense oligonucleotide comprises a modification.

[0200] In some embodiments, a method for increasing 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 a sequence set forth 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 13-30 nucleotides. In some embodiments, the antisense oligonucleotide comprises a modification.

[0201] In some embodiments, a method for increasing 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 set forth in SEQ ID NO: 1, and wherein the antisense oligonucleotide comprises a sequence set forth in any one of SEQ ID NOs: 961-983 and 1116-1145. In some embodiments, the antisense oligonucleotide consists of 13 nucleotides complementary to the sequence set forth in SEQ ID NO: 1, and wherein the antisense oligonucleotide consists of a sequence set forth in any one of SEQ ID NOs: 961-983 and 1116-1145. In some embodiments, wherein the antisense oligonucleotide is provided to a cell, expression of UNC13A mRNA is increased in the cell by 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 a wild-type cell. In some embodiments, expression of UNC13A mRNA is increased in cells 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, expression of UNC13A mRNA is increased in cells 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, in which an antisense oligonucleotide is provided to a cell, expression of UNC13A cryptic exon mRNA is reduced in cells 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 in which an antisense oligonucleotide is provided to a cell, expression of UNC13A protein is increased in the cell by 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 a wild-type cell.In some embodiments, expression of UNC13A protein is increased in cells by between about 10% and about 95%, 20% and about 85%, 30% and about 75%, 40% and about 65%, or 50% and about 55% of the level of UNC13A protein expressed in wild-type cells. In some embodiments, expression of UNC13A protein is increased in cells by between about 25% and about 1000%, about 50% and about 800%, about 100% and about 700%, about 200% and about 600%, or about 300% and about 500%. In some embodiments in which an antisense oligonucleotide is provided to a cell, 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 potentials, neuronal synaptic plasticity, and neuronal longevity. In some embodiments, the antisense oligonucleotide consists of the sequence set forth 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 set forth in any one of SEQ ID NOs: 1059-1074, 1075-1089, and 1090-1103. In some embodiments, the antisense oligonucleotide consists of the sequence set forth 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 oligonucleotides, 2'O-methoxyethyl antisense oligonucleotides, phosphorothioate antisense oligonucleotides, and LNA antisense oligonucleotides.

[0202] In some embodiments, a method for increasing 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 set forth in SEQ ID NO: 1, wherein the antisense oligonucleotide comprises a sequence set forth in any one of SEQ ID NOs: 984-1004 and 1146-1173. In some embodiments, the antisense oligonucleotide consists of 15 nucleotides complementary to the sequence set forth in SEQ ID NO: 1, wherein the antisense oligonucleotide consists of a sequence set forth in any one of SEQ ID NOs: 984-1004 and 1146-1173. In some embodiments, wherein the antisense oligonucleotide is provided to a cell, expression of UNC13A mRNA is increased in the cell by 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 a wild-type cell. In some embodiments, expression of UNC13A mRNA is increased in cells 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, expression of UNC13A mRNA is increased in cells 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, in which an antisense oligonucleotide is provided to a cell, expression of UNC13A cryptic exon mRNA is reduced in cells 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 in which an antisense oligonucleotide is provided to a cell, expression of UNC13A protein is increased in the cell by 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 a wild-type cell.In some embodiments, expression of UNC13A protein is increased in cells by between about 10% and about 95%, 20% and about 85%, 30% and about 75%, 40% and about 65%, or 50% and about 55% of the level of UNC13A protein expressed in wild-type cells. In some embodiments, expression of UNC13A protein is increased in cells by between about 25% and about 1000%, about 50% and about 800%, about 100% and about 700%, about 200% and about 600%, or about 300% and about 500%. In some embodiments in which an antisense oligonucleotide is provided to a cell, 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 potentials, neuronal synaptic plasticity, and neuronal longevity. In some embodiments, the antisense oligonucleotide consists of the sequence set forth 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 set forth in any one of SEQ ID NOs: 1059-1074, 1075-1089, and 1090-1103. In some embodiments, the antisense oligonucleotide consists of the sequence set forth 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 oligonucleotides, 2'O-methoxyethyl antisense oligonucleotides, phosphorothioate antisense oligonucleotides, and LNA antisense oligonucleotides.

[0203] In some embodiments, a method for increasing expression of UNC13A protein in a cell by at least 10% comprises contacting a cell with an antisense oligonucleotide, wherein the antisense oligonucleotide comprises 16 nucleotides complementary to the sequence set forth in SEQ ID NO: 1, and wherein the antisense oligonucleotide is selected from the group consisting of SEQ ID NOs: 41, 42, 56, 57, 71, 72, 86, 87, 101, 102, 116, 117, 131, 132, 146, 147, 16 1, 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 set forth in SEQ ID NO: 1, and the antisense oligonucleotide consists of 16 nucleotides complementary to the sequence set forth in 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 a cell, expression of UNC13A mRNA is increased in the cell by 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 a wild-type cell.In some embodiments, expression of UNC13A mRNA is increased in cells 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, expression of UNC13A mRNA is increased in cells 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, in which an antisense oligonucleotide is provided to a cell, expression of UNC13A cryptic exon mRNA is reduced in cells 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, in which an antisense oligonucleotide is provided to a cell, expression of UNC13A protein is increased in the cell by 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 a wild-type cell. In some embodiments, expression of UNC13A protein is increased in the cell by between about 10% and about 95%, 20% and about 85%, 30% and about 75%, 40% and about 65%, or 50% and about 55% of the level of UNC13A protein expressed in a wild-type cell. In some embodiments, expression of UNC13A protein is increased in the cell by between about 25% and about 1000%, about 50% and about 800%, about 100% and about 700%, about 200% and about 600%, or about 300% and about 500%. In some embodiments, the antisense oligonucleotide is provided to a cell, and 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 cell morphology, neurite length, neurotransmitter release, action potentials, neuronal synaptic plasticity, and neuronal longevity. In some embodiments, the antisense oligonucleotide consists of the sequence set forth 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 set forth in any one of SEQ ID NOs: 1059-1074, 1075-1089, and 1090-1103. In some embodiments, the antisense oligonucleotide consists of the sequence set forth 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 oligonucleotides, 2'O-methoxyethyl antisense oligonucleotides, phosphorothioate antisense oligonucleotides, and LNA antisense oligonucleotides.

[0204] In some embodiments, a method for increasing expression of UNC13A protein in a cell by at least 10% comprises contacting a cell with an antisense oligonucleotide, wherein the antisense oligonucleotide comprises 17 nucleotides complementary to the sequence set forth in SEQ ID NO: 1, and wherein the antisense oligonucleotide is selected from the group consisting of SEQ ID NOs: 43, 58, 73, 88, 103, 118, 133, 148, 163, 178, 19 3, 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 to 1023, and 1174 to 1199. In some embodiments, the antisense oligonucleotide consists of 17 nucleotides complementary to the sequence set forth in SEQ ID NO: 1, and the antisense oligonucleotide consists of any one of the sequences set forth in 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, in which an antisense oligonucleotide is provided to a cell, expression of UNC13A mRNA is increased in the cell by 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 a wild-type cell. In some embodiments, expression of UNC13A mRNA is increased in the cell by between about 10% and about 95%, 20% and about 85%, 30% and about 75%, 40% and about 65%, or 50% and about 55% of the level of UNC13A mRNA expressed in a wild-type cell.In some embodiments, expression of UNC13A mRNA is increased in the cells 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, when an antisense oligonucleotide is provided to the cells, expression of UNC13A cryptic exon mRNA is decreased in the cells 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, when an antisense oligonucleotide is provided to the cells, expression of UNC13A protein is increased in the cells by 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, expression of UNC13A protein is increased in cells by between about 10% and about 95%, 20% and about 85%, 30% and about 75%, 40% and about 65%, or 50% and about 55% of the level of UNC13A protein expressed in wild-type cells. In some embodiments, expression of UNC13A protein is increased in cells by between about 25% and about 1000%, about 50% and about 800%, about 100% and about 700%, about 200% and about 600%, or about 300% and about 500%. In some embodiments in which an antisense oligonucleotide is provided to a cell, 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 potentials, neuronal synaptic plasticity, and neuronal longevity. In some embodiments, the antisense oligonucleotide consists of the sequence set forth 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 set forth in any one of SEQ ID NOs: 1059-1074, 1075-1089, and 1090-1103.In some embodiments, the antisense oligonucleotide consists of 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 oligonucleotides, 2'O-methoxyethyl antisense oligonucleotides, phosphorothioate antisense oligonucleotides, and LNA antisense oligonucleotides.

[0205] In some embodiments, a method for increasing 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 set forth in SEQ ID NO: 1, and wherein the antisense oligonucleotide is selected from the group consisting of SEQ ID NOs: 38-40, 44, 45, 59, 60, 74, 75, 89, 90, 104, 105, 119, 120, 121, 122, 123, 124, 125, 126, 127, 128, 129, 130, 131, 132, 133, 134, 135, 136, 137, 138, 139, 140, 141, 142, 143, 144, 145, 146, 147, 148, 149, 150, 151, 152, 153, 154, 155, 156, 157, 158, 159, 160, 161, 162, 163, 164, 165, 166, 167, 168, 169, 170, 171, 172, 173, 174, 175, 176, 177, 178, 179, 180, 181, 182, 183, 184, 185, 186, 187, 188, 189, 190, 191, 192, 193, 34, 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 to 638, 817, 818, 845, 855, 871, 882, 904, 958, 959, 960, 1024 to 1041, and 1200 to 1224. In some embodiments, the antisense oligonucleotide consists of 18 nucleotides complementary to the sequence set forth in SEQ ID NO: 1, and the antisense oligonucleotide is selected from the group consisting 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, 190, 200, 210, 212, 214, 216, 218, 220, 222, 224, 226, 228, 230, 232, 234, 235, 249, 150, 164, 165, 179, 180, 191, 219, 225, 232, 233, 234, 235, 249, 150, 164, 165, 179, 180, 192, 235, 246, 247, 248, 250, 252, 254, 256, 258, 260, 262, 264, 265, 270, 272, 274, 275, 276, 278, 279, 280, 281, 282, 283, 284, 285, 286, 287, 288, 289, 300, 301, 302, 303, 304, 305, 4, 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 a cell, expression of UNC13A mRNA is increased in the cell by 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 a wild-type cell. In some embodiments, expression of UNC13A mRNA is increased in cells by between about 10% and about 95%, 20% and about 85%, 30% and about 75%, 40% and about 65%, or 50% and about 55% of the level of UNC13A mRNA expressed in wild-type cells.In some embodiments, expression of UNC13A mRNA is increased in the cells 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, when an antisense oligonucleotide is provided to the cells, expression of UNC13A cryptic exon mRNA is decreased in the cells 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, when an antisense oligonucleotide is provided to the cells, expression of UNC13A protein is increased in the cells by 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, expression of UNC13A protein is increased in cells by between about 10% and about 95%, 20% and about 85%, 30% and about 75%, 40% and about 65%, or 50% and about 55% of the level of UNC13A protein expressed in wild-type cells. In some embodiments, expression of UNC13A protein is increased in cells by between about 25% and about 1000%, about 50% and about 800%, about 100% and about 700%, about 200% and about 600%, or about 300% and about 500%. In some embodiments in which an antisense oligonucleotide is provided to a cell, 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 potentials, neuronal synaptic plasticity, and neuronal longevity. In some embodiments, the antisense oligonucleotide consists of the sequence set forth 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 set forth in any one of SEQ ID NOs: 1059-1074, 1075-1089, and 1090-1103.In some embodiments, the antisense oligonucleotide consists of 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 oligonucleotides, 2'O-methoxyethyl antisense oligonucleotides, phosphorothioate antisense oligonucleotides, and LNA antisense oligonucleotides.

[0206] In some embodiments, a method for increasing 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 set forth in SEQ ID NO: 1, and wherein the antisense oligonucleotide comprises a sequence set forth 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 19 nucleotides complementary to the sequence set forth in SEQ ID NO: 1, and the antisense oligonucleotide consists of the sequence set forth 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, where an antisense oligonucleotide is provided to a cell, expression of UNC13A mRNA is increased in the cell by 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 a wild-type cell. In some embodiments, expression of UNC13A mRNA is increased in the cell by between about 10% and about 95%, 20% and about 85%, 30% and about 75%, 40% and about 65%, or 50% and about 55% of the level of UNC13A mRNA expressed in a wild-type cell. In some embodiments, expression of UNC13A mRNA is increased in the cell by between about 25% and about 1000%, about 50% and about 800%, about 100% and about 700%, about 200% and about 600%, or about 300% and about 500%.In some embodiments, where an antisense oligonucleotide is provided to a cell, expression of UNC13A cryptic exon mRNA is reduced in the cell 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, expression of UNC13A protein is increased in the cell by 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, expression of UNC13A protein is increased in the cell by between 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, expression of UNC13A protein is increased in cells 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, in which an antisense oligonucleotide is provided to a cell, 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 cell morphology, neurite length, neurotransmitter release, action potentials, neuronal synaptic plasticity, and neuronal longevity. In some embodiments, the antisense oligonucleotide consists of the sequence set forth 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 set forth in any one of SEQ ID NOs: 1059-1074, 1075-1089, and 1090-1103. In some embodiments, the antisense oligonucleotide consists of the sequence set forth 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 oligonucleotides, 2'O-methoxyethyl antisense oligonucleotides, phosphorothioate antisense oligonucleotides, and LNA antisense oligonucleotides.

[0207] In some embodiments, a method for increasing expression of UNC13A protein in a cell by at least 10% comprises contacting a cell with an antisense oligonucleotide, wherein the antisense oligonucleotide comprises 20 nucleotides complementary to the sequence set forth in SEQ ID NO: 1, and wherein the antisense oligonucleotide comprises a sequence set forth 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 20 nucleotides complementary to the sequence set forth in SEQ ID NO: 1, and the antisense oligonucleotide consists of the sequence set forth 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, in which an antisense oligonucleotide is provided to a cell, expression of UNC13A mRNA is increased in the cell by 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 a wild-type cell. In some embodiments, expression of UNC13A mRNA is increased in the cell by between about 10% and about 95%, 20% and about 85%, 30% and about 75%, 40% and about 65%, or 50% and about 55% of the level of UNC13A mRNA expressed in a wild-type cell.In some embodiments, expression of UNC13A mRNA is increased in the cells 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, when an antisense oligonucleotide is provided to the cells, expression of UNC13A cryptic exon mRNA is decreased in the cells 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, when an antisense oligonucleotide is provided to the cells, expression of UNC13A protein is increased in the cells by 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, expression of UNC13A protein is increased in cells by between about 10% and about 95%, 20% and about 85%, 30% and about 75%, 40% and about 65%, or 50% and about 55% of the level of UNC13A protein expressed in wild-type cells. In some embodiments, expression of UNC13A protein is increased in cells by between about 25% and about 1000%, about 50% and about 800%, about 100% and about 700%, about 200% and about 600%, or about 300% and about 500%. In some embodiments in which an antisense oligonucleotide is provided to a cell, 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 potentials, neuronal synaptic plasticity, and neuronal longevity. In some embodiments, the antisense oligonucleotide consists of the sequence set forth 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 set forth in any one of SEQ ID NOs: 1059-1074, 1075-1089, and 1090-1103.In some embodiments, the antisense oligonucleotide consists of 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 oligonucleotides, 2'O-methoxyethyl antisense oligonucleotides, phosphorothioate antisense oligonucleotides, and LNA antisense oligonucleotides.

[0208] In some embodiments, a method for increasing expression of UNC13A protein in a cell by at least 10% comprises contacting a cell with an antisense oligonucleotide, wherein the antisense oligonucleotide comprises 21 nucleotides complementary to the sequence set forth in SEQ ID NO: 1, and wherein the antisense oligonucleotide comprises a sequence set forth 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 set forth in SEQ ID NO: 1, wherein the antisense oligonucleotide consists of the sequence set forth 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 a cell, expression of UNC13A mRNA is increased in the cell by 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 a wild-type cell. In some embodiments, expression of UNC13A mRNA is increased in cells 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, expression of UNC13A mRNA is increased in cells 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, expression of UNC13A cryptic exon mRNA is reduced in the cell 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, expression of UNC13A protein is increased in the cell by 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, expression of UNC13A protein is increased in the cell by between 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, expression of UNC13A protein is increased in cells 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, in which an antisense oligonucleotide is provided to a cell, 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 cell morphology, neurite length, neurotransmitter release, action potentials, neuronal synaptic plasticity, and neuronal longevity. In some embodiments, the antisense oligonucleotide consists of the sequence set forth 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 set forth in any one of SEQ ID NOs: 1059-1074, 1075-1089, and 1090-1103. In some embodiments, the antisense oligonucleotide consists of the sequence set forth 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 oligonucleotides, 2'O-methoxyethyl antisense oligonucleotides, phosphorothioate antisense oligonucleotides, and LNA antisense oligonucleotides.

[0209] In some embodiments, the method of increasing 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 set forth in SEQ ID NO: 1, and wherein the antisense oligonucleotide is selected from the group consisting of SEQ ID NOs: 50, 51, 65, 66, 80, 81, 95, 96, 110, 111, 12 5, 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 to 1103, and 1293 to 1313. In some embodiments, the antisense oligonucleotide consists of 22 nucleotides complementary to the sequence set forth in SEQ ID NO: 1, and the antisense oligonucleotide consists of any one of the sequences set forth in 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, in which an antisense oligonucleotide is provided to a cell, expression of UNC13A mRNA is increased in the cell by 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 a wild-type cell. In some embodiments, expression of UNC13A mRNA is increased in the cell by between about 10% and about 95%, 20% and about 85%, 30% and about 75%, 40% and about 65%, or 50% and about 55% of the level of UNC13A mRNA expressed in a wild-type cell.In some embodiments, expression of UNC13A mRNA is increased in the cells 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, when an antisense oligonucleotide is provided to the cells, expression of UNC13A cryptic exon mRNA is decreased in the cells 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, when an antisense oligonucleotide is provided to the cells, expression of UNC13A protein is increased in the cells by 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, expression of UNC13A protein is increased in cells by between about 10% and about 95%, 20% and about 85%, 30% and about 75%, 40% and about 65%, or 50% and about 55% of the level of UNC13A protein expressed in wild-type cells. In some embodiments, expression of UNC13A protein is increased in cells by between about 25% and about 1000%, about 50% and about 800%, about 100% and about 700%, about 200% and about 600%, or about 300% and about 500%. In some embodiments in which an antisense oligonucleotide is provided to a cell, 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 potentials, neuronal synaptic plasticity, and neuronal longevity. In some embodiments, the antisense oligonucleotide consists of the sequence set forth in any one of SEQ ID NOs: 1059-1074, 1075...

Claims

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

1.

2. 1. A method for increasing expression of UNC13A protein in a cell by at least 10%, comprising contacting the cell with an antisense oligonucleotide comprising 13 to 30 nucleotides, wherein the antisense oligonucleotide is complementary to a sequence set forth in any one of SEQ ID NOs: 1339-1342 or a portion thereof.

3. 3. The method of claim 1 or 2, wherein the antisense oligonucleotide comprises a sequence set forth in any one of SEQ ID NOs: 20-355, 400-638, 810-957, or 961-1332.

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

1.

5. 5. The method of 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 the antisense oligonucleotide is complementary to the sequence set forth in SEQ ID NO:

1.

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

1.

7. 6. The method of any one of claims 1 to 5, wherein the antisense oligonucleotide consists of 13 nucleotides complementary to the sequence set forth in SEQ ID NO: 1, and the antisense oligonucleotide consists of a sequence set forth in any one of SEQ ID NOs: 961 to 983 and 1116 to 1145.

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

9. The antisense oligonucleotide consists of 16 nucleotides complementary to the sequence set forth in SEQ ID NO: 1, and the antisense oligonucleotide is selected from the group consisting 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, 238, 239, 240, 241, 242, 243, 244, 245, 246, 247, 248, 249, 250, 251, 252, 253, 254, 255, 256, 257, 258, 259, 260, 261, 262, 263, 264, 265, 266, 267, 268, 269, 270, 271, 272, 273, 274, 275, 276, 277, 278, 279, 280, 281, 282, 283, 284, 285, 286, 287, 288, 289, 290, 300, 301, 302, 303, 304, 305, 306, 307, 308, 309, 310, 311, 312, 3 5. The method of claim 1, wherein the sequence comprises any one of the sequences listed in any one of: 51, 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 antisense oligonucleotide consists of 17 nucleotides complementary to the sequence set forth in SEQ ID NO: 1, and the antisense oligonucleotide is selected from the group consisting 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, 330, 331, 332, 333, 334, 335, 336, 337, 338, 339, 340, 341, 342, 343, 344, 345, 346, 347, 348, 349, 350, 351, 352, 353, 354, 355, 356, 357, 358, 359, 360, 361, 362, 363, 364, 365, 366, 367, 368, 369, 370, 371, 372, 373, 374, 375, 376, 377, 378, 379, 380, 381, 382, ​​383, 384, 385, 386, 387, 388, 389, 390, 400, 401, 402, 403, 404, 405, 406, 407, 408, 409, 41 6. The method of any one of claims 1 to 5, wherein the sequence comprises any one of the sequences set forth in any one of: 42, 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 to 1023, and 1174 to 1199.

11. The antisense oligonucleotide consists of 18 nucleotides complementary to the sequence set forth in SEQ ID NO: 1, and the antisense oligonucleotide is selected from the group consisting of SEQ ID NOs: 38 to 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, 6. The method of any one of claims 1 to 5, wherein the sequence comprises any one of the sequences set forth in any one of: 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. 7. The method of any one of claims 1 to 6, wherein the antisense oligonucleotide consists of 19 nucleotides complementary to the sequence set forth in SEQ ID NO:1, and wherein the antisense oligonucleotide consists of the sequence set forth 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.

13. 7. The method of any one of claims 1 to 6, wherein the antisense oligonucleotide consists of 20 nucleotides complementary to the sequence set forth in SEQ ID NO: 1, and the antisense oligonucleotide consists of the sequence set forth 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.

14. 7. The method of any one of claims 1 to 6, wherein the antisense oligonucleotide consists of 21 nucleotides complementary to the sequence set forth in SEQ ID NO: 1, and the antisense oligonucleotide consists of the sequence set forth 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.

15. 7. The method of any one of claims 1 to 6, wherein the antisense oligonucleotide consists of 22 nucleotides complementary to the sequence set forth in SEQ ID NO: 1, and the antisense oligonucleotide consists of the sequence set forth 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.

16. 6. The method of any one of claims 1 to 5, wherein the antisense oligonucleotide consists of 23 nucleotides complementary to the sequence set forth in SEQ ID NO:1, and wherein the antisense oligonucleotide consists of the sequence set forth 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.

17. The antisense oligonucleotide consists of 24 nucleotides complementary to the sequence set forth in SEQ ID NO: 1, and the antisense oligonucleotide is selected from the group consisting of SEQ ID NOs: 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, 234 821, 822, 832-848, 858, 874, 887, 907, 919, 920, 923, 931, 946-951, 1104-1115, and 1314-1332.

18. The antisense oligonucleotide consists of 25 nucleotides complementary to the sequence set forth in SEQ ID NO: 1, and the antisense oligonucleotide is selected from the group consisting of SEQ ID NOs: 55, 70, 85, 100, 115, 130, 145, 160, 175, 190, 205, 220, 235, 250, 265, 280, 295, 310, 324, 325, 339, 340, 354 6. The method of any one of claims 1 to 5, wherein the sequence comprises any one of the sequences set forth in any one of the following: 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. 19. The method of any one of claims 1 to 18, wherein the antisense oligonucleotide is complementary to a sequence set forth in any one of SEQ ID NOs: 700 to 703.

20. 20. The method of 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 to a sequence set forth in any one of SEQ ID NOs: 700-703.

21. 21. The method of 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 to the sequence set forth in SEQ ID NO:

701.

22. 22. The method of 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 to the sequence set forth in SEQ ID NO:

703.

23. 23. The method of any one of claims 1 to 22, wherein the antisense oligonucleotide is complementary to a sequence set forth in any one of SEQ ID NOs: 1339 to 1342 or a portion thereof.

24. 24. The method of any one of claims 1 to 21 and 23, wherein the antisense oligonucleotide is complementary to the sequence set forth in SEQ ID NO: 1339 or a portion thereof.

25. 25. The method of any one of claims 1 to 21 and 23 to 24, wherein the antisense oligonucleotide is complementary to the sequence set forth in SEQ ID NO: 1340 or a portion thereof.

26. 24. The method of any one of claims 1 to 20 and 22 to 23, wherein the antisense oligonucleotide is complementary to the sequence set forth in SEQ ID NO: 1341 or a portion thereof.

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

28. 28. The method of 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 set forth in any one of SEQ ID NOs: 1339-1342.

29. 29. The method of 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 a sequence set forth in any one of SEQ ID NOs: 1339-1342.

30. 30. The method of 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 set forth in SEQ ID NO: 1339.

31. 30. The method of 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 set forth in SEQ ID NO: 1340.

32. 30. The method of 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 set forth in SEQ ID NO: 1341.

33. 30. The method of claim 29, wherein 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 set forth in SEQ ID NO: 1342.

34. 34. The method of any one of claims 1 to 33, wherein the cells express low levels of UNC13A prior to contacting the cells with the antisense oligonucleotide.

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

36. 36. The method of claim 35, wherein UNC13A protein levels are measured by Western blot, imaging, ELISA, fluorescent reporter assay, luminescence assay, or immunohistochemistry.

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

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

39. 39. The method of 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 to the sequence set forth in SEQ ID NO:

1.

40. 40. The method of any one of claims 1 to 39, wherein the antisense oligonucleotide restores 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 a wild-type cell.

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

42. The method of claim 40 or 41, wherein the antisense oligonucleotide consists of a sequence set forth in any one of SEQ ID NOs: 1059-1074, 1075-1089, 1090-1103, 1248-1270, 1271-1292, and 1293-1313.

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

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

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

46. The method of claim 44 or 45, wherein the antisense oligonucleotide consists of a sequence set forth in any one of SEQ ID NOs: 1059-1074, 1075-1089, 1090-1103, 1248-1270, 1271-1292, and 1293-1313.

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

9.

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

1.

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

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

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

52. 52. The method of any one of claims 1 to 51, wherein the antisense oligonucleotide restores 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 a wild-type cell.

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

54. 54. The method of claim 52 or 53, wherein the antisense oligonucleotide consists of a sequence set forth in any one of SEQ ID NOs: 1059-1074, 1075-1089, 1090-1103, 1248-1270, 1271-1292, and 1293-1313.

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

56. The method of any one of claims 1 to 6, 13 to 15, and 19 to 55, wherein the antisense oligonucleotide consists of a sequence set forth in any one of SEQ ID NOs: 1059-1074, 1075-1089, 1090-1103, 1248-1270, 1271-1292, and 1293-1313.

57. The method of any one of claims 1 to 6, 13 to 15, and 19 to 56, wherein the antisense oligonucleotide consists of a sequence set forth in any one of SEQ ID NOs: 1059 to 1074, 1075 to 1089, and 1090 to 1103.

58. The method of any one of claims 1 to 6, 13 to 15, and 19 to 56, wherein the antisense oligonucleotide consists of a sequence set forth in any one of SEQ ID NOs: 1248 to 1270, 1271 to 1292, and 1293 to 1313.

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

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

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

62. 62. The method of claim 61, wherein the risk allele is selected from the group consisting of rsl2608932 (hg38 chr19:17,641,880 A→C), rsl2973192 (hg38 chr19:17,642,430 C→G), rs56041637 (hg38 chr19:17,642,033-17,642,056 0-2 CATC repeats→3-5 CATC repeats), and rs62121687 (hg38 chr19:17,642,351 C→A), or any combination thereof.

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

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

65. 65. The method of any one of claims 1 to 64, wherein the cells express low levels of TDP-43 protein.

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

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

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

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

1.

70. 1. A method of treating amyotrophic lateral sclerosis (ALS) or frontotemporal dementia (FTD) in an individual, comprising administering to the individual an antisense oligonucleotide comprising 13-30 nucleotides, wherein the antisense oligonucleotide is complementary to the sequence set forth in SEQ ID NO:1, and 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.

71. 71. The method of claim 70, wherein the risk allele is selected from the group consisting of rsl2608932 (hg38 chr19:17,641,880 A→C), rsl2973192 (hg38 chr19:17,642,430 C→G), rs56041637 (hg38 chr19:17,642,033-17,642,056 0-2 CATC repeats→3-5 CATC repeats), and rs62121687 (hg38 chr19:17,642,351 C→A), or any combination thereof.

72. 71. The method of claim 70, wherein the ALS or FTD associated mutation is a mutation in TDP43.

73. 73. The method of any one of claims 1 to 72, wherein the antisense oligonucleotide has a sequence that is complementary to a 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.

74. 74. The method of 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 that regulates UNC13A splicing, wherein the antisense oligonucleotide has a sequence that is complementary to a sequence of UNC13A mRNA located between the 5' splicing donor site and the 3' splicing acceptor site of a cryptic exon located between exons 20 and 21 of the UNC13A gene.

76. An antisense oligonucleotide that modulates UNC13A splicing, comprising 13 to 30 nucleotides, wherein the antisense oligonucleotide comprises a sequence set forth in any one of SEQ ID NOs: 20-355, 400-638, 810-959, or 961-1332.

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

1.

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

79. 79. The antisense oligonucleotide of any one of claims 75 to 78, comprising at least 80%, at least 90%, at least 95%, at least 98%, or 100% complementarity to a sequence set forth in any one of SEQ ID NOs: 700 to 703.

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

701.

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

703.

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

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

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

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

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

87. The antisense oligonucleotide of 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 a sequence set forth in any one of SEQ ID NOs: 1339 to 1342.

88. The antisense oligonucleotide of 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 a sequence set forth in any one of SEQ ID NOs: 1339 to 1342.

89. The antisense oligonucleotide of 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 set forth in SEQ ID NO: 1339.

90. The antisense oligonucleotide of 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 set forth in SEQ ID NO: 1340.

91. The antisense oligonucleotide of 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 set forth in SEQ ID NO: 1341.

92. The antisense oligonucleotide of 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 set forth in SEQ ID NO: 1342.

93. An antisense oligonucleotide comprising a sequence set forth in any one of SEQ ID NOs: 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 to 30 nucleotides.

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

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

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

1.

99. An antisense oligonucleotide consisting of a sequence set forth in any one of SEQ ID NOs: 20 to 355, 400 to 638, 810 to 959, or 961 to 1332.

100. An antisense oligonucleotide, wherein the antisense oligonucleotide is complementary to a sequence set forth in any one of SEQ ID NOs: 1339 to 1342 or a portion thereof.

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

102. The antisense oligonucleotide of any one of claims 75 to 96 and 98 to 100, wherein the antisense oligonucleotide consists of 15 nucleotides that are complementary to the sequence set forth in SEQ ID NO: 1, and the antisense oligonucleotide consists of the sequence set forth in any one of SEQ ID NOs: 984 to 1004 and 1146 to 1173.

103. The antisense oligonucleotide consists of 16 nucleotides complementary to the sequence set forth in SEQ ID NO: 1, and the antisense oligonucleotide is selected from the group consisting 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, 270, 272, 274, 276, 278, 279, 280, 281, 282, 283, 284, 285, 286, 287, 288, 289, 290, 300, 301, 302, 303, 304, 305, 306, 307, 308, 309, 310, 311, 312, 313, 314, 315, 316, 317, 318, 319, 320, 321, 322, 323, 324, 325, 326, 327, 328, 329, 330, 331, 332, 333, 334, 335, 336, 337, 338, 339, 340, 341, 342, 343, 344, 3 67, 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. The antisense oligonucleotide of any one of claims 75 to 96 and 98 to 100, comprising the sequence set forth in any one of the following:

104. The antisense oligonucleotide consists of 17 nucleotides complementary to the sequence set forth in SEQ ID NO: 1, and the antisense oligonucleotide is selected from the group consisting 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, 817, 818, 819, 820, 821, 822, 823, 824, 825, 826, 827, 828, 829, 830, 831, 832, 833, 834, 835, 836, 837, 838, 839, 840, 841, 842, 843, 844, 845, 846, 847, 848, 849, 850, 851, 852, 853, 854, 855, 856, 857, 858, 859, 860, 861, 862, 863, 864, 865, 866, 867, 868, 870, 871, 872, 873, 874, 875, 876, 877, 878, 879, 880, 881, 882, 883, 88 The antisense oligonucleotide according to any one of claims 75 to 96 and 98 to 100, comprising a sequence set forth in any one of 15, 816, 828, 829, 836, 839, 843, 844, 853, 854, 862, 865, 869, 870, 879, 880, 881, 891, 896, 897, 902, 903, 1005 to 1023, and 1174 to 1199.

105. The antisense oligonucleotide consists of 18 nucleotides complementary to the sequence set forth in SEQ ID NO: 1, and the antisense oligonucleotide is selected from the group consisting of SEQ ID NOs: 38 to 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. The antisense oligonucleotide according to any one of claims 75 to 96 and 98 to 100, comprising a sequence set forth in any one of the following:

106. The antisense oligonucleotide of any one of claims 75 to 100, wherein the antisense oligonucleotide consists of 19 nucleotides that are complementary to the sequence set forth in SEQ ID NO: 75, and the antisense oligonucleotide consists of the sequence set forth 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. 101. The antisense oligonucleotide of any one of claims 75 to 100, wherein the antisense oligonucleotide consists of 20 nucleotides complementary to the sequence set forth in SEQ ID NO: 1, and wherein the antisense oligonucleotide consists of the sequence set forth 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 of any one of claims 75 to 100, wherein the antisense oligonucleotide consists of 21 nucleotides complementary to the sequence set forth in SEQ ID NO: 1, and the antisense oligonucleotide consists of a sequence set forth 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 consists of 22 nucleotides complementary to the sequence set forth in SEQ ID NO: 1, and the antisense oligonucleotide is selected from the group consisting 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, 217, 218, 220, 221, 222, 223, 224, 225, 226, 227, 228, 229, 230, 231, 232, 233, 234, 235, 236, 237, 238, 239, 240, 241, 242, 243, 244, 245, 246, 247, 248, 249, 250, 251, 252, 253, 254, 255, 256, 257, 258, 259, 260, 261, 262, 263, 264, 265, 266, 267, 268, 269, 270, 271, 272, 273, 274, 275, 276, 277, 278, 279, 280, 281, 282, 283, 284, 285, 286, 287, 2 The antisense oligonucleotide according to any one of claims 75 to 100, comprising a sequence set forth in any one of: 30, 231, 245, 246, 260, 261, 275, 276, 290, 291, 305, 306, 320, 335, 350, 912, 913, 914, 938, 939, 940, 941, 1090 to 1103, and 1293 to 1313.

110. 101. The antisense oligonucleotide of any one of claims 75 to 96 and 98 to 100, wherein the antisense oligonucleotide consists of 23 nucleotides that are complementary to the sequence set forth in SEQ ID NO: 1, and wherein the antisense oligonucleotide consists of the sequence set forth 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. The antisense oligonucleotide consists of 24 nucleotides complementary to the sequence set forth in SEQ ID NO: 1, and the antisense oligonucleotide is selected from the group consisting of SEQ ID NOs: 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. An antisense oligonucleotide according to any one of claims 75 to 96 and 98 to 100, comprising the sequence set forth in any one of

112. The antisense oligonucleotide consists of 25 nucleotides complementary to the sequence set forth in SEQ ID NO: 1, and the antisense oligonucleotide is selected from the group consisting of SEQ ID NOs: 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 ... 4, 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. The antisense oligonucleotide of any one of claims 75 to 96 and 98 to 100,

113. The antisense oligonucleotide of any one of claims 75 to 80 and 82 to 112, wherein the antisense oligonucleotide is complementary to the sequence set forth in SEQ ID NO: 1339 or a portion thereof.

114. The antisense oligonucleotide of any one of claims 75 to 80 and 82 to 113, wherein the antisense oligonucleotide is complementary to the sequence set forth in SEQ ID NO: 1340 or a portion thereof.

115. The antisense oligonucleotide of any one of claims 75 to 79 and 81 to 112, wherein the antisense oligonucleotide is complementary to the sequence set forth in SEQ ID NO: 1341 or a portion thereof.

116. 116. The antisense oligonucleotide of any one of claims 75 to 79, 81 to 112, and 115, wherein the antisense oligonucleotide is complementary to the sequence set forth in SEQ ID NO: 1342 or a portion thereof.

117. The antisense oligonucleotide of 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 a sequence set forth in any one of SEQ ID NOs: 1339 to 1342.

118. The antisense oligonucleotide of 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 a sequence set forth in any one of SEQ ID NOs: 1339 to 1342.

119. The antisense oligonucleotide of 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 set forth in SEQ ID NO: 1339.

120. The antisense oligonucleotide of 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 set forth in SEQ ID NO: 1340.

121. The antisense oligonucleotide of 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 set forth in SEQ ID NO: 1341.

122. The antisense oligonucleotide of 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 set forth in SEQ ID NO: 1342.

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

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

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

126. 126. The antisense oligonucleotide of any one of claims 75 to 125, wherein the 5' end and / or the 3' end of the antisense oligonucleotide comprises a modification, and 2, 3, 4, or 5 nucleotides are modified at each of the 5' end and / or the 3' end of the antisense oligonucleotide.

127. 127. The antisense oligonucleotide of 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 a modification.

128. 128. The antisense oligonucleotide of any one of claims 75 to 127, wherein the modification is 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. The antisense oligonucleotide of any one of claims 75 to 128, wherein the modification is a 2'OMe antisense oligonucleotide.

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

131. 131. The pharmaceutical composition of claim 130, comprising an excipient and / or a buffer.

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

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