{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"institution":[{"name":"bioRxiv"}],"indexed":{"date-parts":[[2026,1,16]],"date-time":"2026-01-16T14:05:30Z","timestamp":1768572330157,"version":"3.49.0"},"posted":{"date-parts":[[2024,10,12]]},"group-title":"Genetics","reference-count":18,"publisher":"openRxiv","license":[{"start":{"date-parts":[[2024,10,12]],"date-time":"2024-10-12T00:00:00Z","timestamp":1728691200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/publicdomain\/zero\/1.0\/"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"accepted":{"date-parts":[[2024,10,12]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n                <jats:p>\n                  Amyotrophic lateral sclerosis 4 (ALS4) is an autosomal dominant motor neuron disease that is molecularly characterized by reduced R-loop levels and caused by pathogenic variants in\n                  <jats:italic>senataxin<\/jats:italic>\n                  (\n                  <jats:italic>SETX<\/jats:italic>\n                  ).\n                  <jats:italic>SETX<\/jats:italic>\n                  encodes an RNA\/DNA helicase that resolves three-stranded nucleic acid structures called R-loops. Currently, there are no disease-modifying therapies available for ALS4. Given that\n                  <jats:italic>SETX<\/jats:italic>\n                  is haplosufficient, removing the product of the mutated allele presents a potential therapeutic strategy. We designed a series of siRNAs to selectively target the RNA transcript from the ALS4 allele containing the c.1166T&gt;C mutation (p.Leu389Ser). Transfection of HEK293 cells with siRNA and plasmids encoding either wild-type or mutant (Leu389Ser) epitope tagged SETX revealed that three siRNAs specifically reduced mutant SETX protein levels without affecting the wild-type SETX protein. In ALS4 primary fibroblasts, siRNA treatment silenced the endogenous mutant\n                  <jats:italic>SETX<\/jats:italic>\n                  allele, while sparing the wild-type allele, and restored R-loop levels in patient cells. Our findings demonstrate that mutant\n                  <jats:italic>SETX<\/jats:italic>\n                  , differing from wild-type by a single nucleotide, can be effectively and specifically silenced by RNA interference, highlighting the potential of allele-specific siRNA as a therapeutic approach for ALS4.\n                <\/jats:p>","DOI":"10.1101\/2024.10.11.617871","type":"posted-content","created":{"date-parts":[[2024,10,13]],"date-time":"2024-10-13T01:55:15Z","timestamp":1728784515000},"source":"Crossref","is-referenced-by-count":0,"title":["Allele-specific silencing of a dominant\n                  <i>SETX<\/i>\n                  mutation in familial amyotrophic lateral sclerosis type 4"],"prefix":"10.64898","author":[{"given":"Audrey","family":"Winkelsas","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Athena","family":"Apfel","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Brian","family":"Johnson","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"George","family":"Harmison","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Dongjun","family":"Li","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Vivian","family":"Cheung","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-4994-2472","authenticated-orcid":false,"given":"Christopher","family":"Grunseich","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"54368","reference":[{"key":"2024101600301018000_2024.10.11.617871v1.1","doi-asserted-by":"publisher","DOI":"10.1093\/brain\/122.8.1539"},{"key":"2024101600301018000_2024.10.11.617871v1.2","doi-asserted-by":"publisher","DOI":"10.1086\/421054"},{"key":"2024101600301018000_2024.10.11.617871v1.3","doi-asserted-by":"publisher","DOI":"10.1093\/nar\/25.23.4778"},{"key":"2024101600301018000_2024.10.11.617871v1.4","doi-asserted-by":"publisher","DOI":"10.1534\/genetics.114.167585"},{"key":"2024101600301018000_2024.10.11.617871v1.5","doi-asserted-by":"publisher","DOI":"10.15252\/embj.201696174"},{"key":"2024101600301018000_2024.10.11.617871v1.6","doi-asserted-by":"publisher","DOI":"10.1016\/j.molcel.2017.12.030"},{"key":"2024101600301018000_2024.10.11.617871v1.7","doi-asserted-by":"publisher","DOI":"10.1086\/301769"},{"key":"2024101600301018000_2024.10.11.617871v1.8","doi-asserted-by":"crossref","first-page":"547","DOI":"10.1002\/ana.25681","article-title":"Clinical and molecular aspects of senataxin mutations in amyotrophic lateral sclerosis 4","volume":"87","year":"2020","journal-title":"Ann. 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Target Ther"}],"container-title":[],"original-title":[],"link":[{"URL":"https:\/\/syndication.highwire.org\/content\/doi\/10.1101\/2024.10.11.617871","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,1,15]],"date-time":"2026-01-15T07:14:57Z","timestamp":1768461297000},"score":1,"resource":{"primary":{"URL":"http:\/\/biorxiv.org\/lookup\/doi\/10.1101\/2024.10.11.617871"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2024,10,12]]},"references-count":18,"URL":"https:\/\/doi.org\/10.1101\/2024.10.11.617871","relation":{},"subject":[],"published":{"date-parts":[[2024,10,12]]},"subtype":"preprint"}}