{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"institution":[{"name":"bioRxiv"}],"indexed":{"date-parts":[[2026,10,7]],"date-time":"2026-10-07T18:06:15Z","timestamp":1791396375401,"version":"4.3.3"},"posted":{"date-parts":[[2025,12,3]]},"group-title":"Neuroscience","reference-count":69,"publisher":"openRxiv","license":[{"start":{"date-parts":[[2025,12,3]],"date-time":"2025-12-03T00:00:00Z","timestamp":1764720000000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"T.A.G was supported by the National Institute of General Medical Sciences Predoctoral Basic Biomedical Sciences Research Training Program"},{"name":"UCSD Gene Therapy Initiative Seed Grant"},{"name":"NIH","award":["R01 NS103172"],"award-info":[{"award-number":["R01 NS103172"]}]},{"name":"NIH","award":["R01 HG004659"],"award-info":[{"award-number":["R01 HG004659"]}]},{"name":"A.A.S. was supported by a Biomedical Research Fellowship from the Hartwell Foundation"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"accepted":{"date-parts":[[2025,12,3]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                <jats:p>\n                  Amyotrophic lateral sclerosis (ALS) is a neurodegenerative disorder caused by the selective deterioration of motor neurons in the central nervous system (CNS). A key driver of this pathogenesis is nuclear loss of ALS-associated protein TDP-43, leading to mis-splicing of TDP-43 targets including important neuronal genes\n                  <jats:italic>STMN2<\/jats:italic>\n                  and\n                  <jats:italic>UNC13A<\/jats:italic>\n                  . Here, we have developed a gene therapy strategy for ALS and related TDP-43 proteinopathies, to correct mis-splicing of both\n                  <jats:italic>STMN2<\/jats:italic>\n                  and\n                  <jats:italic>UNC13A<\/jats:italic>\n                  cryptic exons using small nuclear RNAs (snRNAs) encoded from a single vector. We identified promoter sequence elements to increase therapeutic snRNA expression by 10-fold, then further optimized the expression cassette with combinatorial snRNA targeting to rescue multiple cryptic splicing targets. The engineered snRNAs restored normal pre-mRNA processing of both\n                  <jats:italic>STMN2<\/jats:italic>\n                  and\n                  <jats:italic>UNC13A<\/jats:italic>\n                  transcripts despite TDP-43 loss of function, rescuing stathmin-2 protein levels in iPSC derived motor neurons, restoring their axonal regeneration capacity to wild-type levels. In addition, adeno-associated virus (AAV) delivery of the snRNAs to the murine central nervous system in the constitutive cryptic splicing model\n                  <jats:italic>Stmn2<\/jats:italic>\n                  <jats:sup>Hum\u0394GU<\/jats:sup>\n                  fully restored cortical\n                  <jats:italic>Stmn2<\/jats:italic>\n                  pre-mRNA processing, highlighting the utility of snRNAs as a therapeutic modality\n                  <jats:italic>in vivo<\/jats:italic>\n                  . Together, this study demonstrates that snRNAs are a promising and versatile therapeutic strategy for the simultaneous correction of multiple aberrant transcripts affected by cryptic splicing in TDP-43 proteinopathies.\n                <\/jats:p>","DOI":"10.64898\/2025.12.01.691001","type":"posted-content","created":{"date-parts":[[2025,12,8]],"date-time":"2025-12-08T12:45:30Z","timestamp":1765197930000},"source":"Crossref","is-referenced-by-count":7,"title":["Dual-targeting snRNA gene therapy rescues STMN2 and UNC13A splicing in TDP-43 proteinopathies"],"prefix":"10.64898","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-0600-2301","authenticated-orcid":false,"given":"Trent 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