{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,17]],"date-time":"2026-09-17T05:52:16Z","timestamp":1789624336581,"version":"build-2803163510"},"reference-count":37,"publisher":"Frontiers Media SA","license":[{"start":{"date-parts":[[2024,6,17]],"date-time":"2024-06-17T00:00:00Z","timestamp":1718582400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":["frontiersin.org"],"crossmark-restriction":true},"short-container-title":["Front. Mol. Neurosci."],"abstract":"<jats:p>\n                    Deafness-causing deficiencies in\n                    <jats:italic>otoferlin<\/jats:italic>\n                    (\n                    <jats:italic>OTOF<\/jats:italic>\n                    ) have been addressed preclinically using dual adeno-associated virus (AAV)-based approaches. However, timing of transduction, recombination of mRNA, and protein expression with dual hybrid AAV methods methods have not previously been characterized. Here, we have established an\n                    <jats:italic>ex vivo<\/jats:italic>\n                    assay to determine the kinetics of dual-AAV mediated expression of\n                    <jats:italic>OTOF<\/jats:italic>\n                    in hair cells of the mouse utricle. We utilized two different recombinant vectors that comprise DB-OTO, one containing the 5\u2032 portion of\n                    <jats:italic>OTOF<\/jats:italic>\n                    under the control of the hair cell-specific\n                    <jats:italic>Myo15<\/jats:italic>\n                    promoter, and the other the 3\u2032 portion of\n                    <jats:italic>OTOF<\/jats:italic>\n                    . We explored specificity of the\n                    <jats:italic>Myo15<\/jats:italic>\n                    promoter in hair cells of the mouse utricle, established dose response characteristics of DB-OTO\n                    <jats:italic>ex vivo<\/jats:italic>\n                    in an OTOF-deficient mouse model, and demonstrated tolerability of AAV1 in utricular hair cells. Furthermore, we established deviations from a one-to-one ratio of 5\u2032 to 3\u2032 vectors with little impact on recombined\n                    <jats:italic>OTOF<\/jats:italic>\n                    . Finally, we established a plateau in quantity of recombined\n                    <jats:italic>OTOF<\/jats:italic>\n                    mRNA and protein expression by 14 to 21\u2009days\n                    <jats:italic>ex vivo<\/jats:italic>\n                    with comparable recovery timing to that\n                    <jats:italic>in vivo<\/jats:italic>\n                    model. These findings demonstrate the utility of an\n                    <jats:italic>ex vivo<\/jats:italic>\n                    model system for exploring expression kinetics and establish\n                    <jats:italic>in vivo<\/jats:italic>\n                    and\n                    <jats:italic>ex vivo<\/jats:italic>\n                    recovery timing of dual AAV-mediated\n                    <jats:italic>OTOF<\/jats:italic>\n                    expression.\n                  <\/jats:p>","DOI":"10.3389\/fnmol.2024.1376128","type":"journal-article","created":{"date-parts":[[2024,6,17]],"date-time":"2024-06-17T08:49:29Z","timestamp":1718614169000},"update-policy":"https:\/\/doi.org\/10.3389\/crossmark-policy","source":"Crossref","is-referenced-by-count":10,"title":["Recovery kinetics of dual AAV-mediated human otoferlin expression"],"prefix":"10.3389","volume":"17","author":[{"given":"Jonathan B.","family":"Sellon","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Kathy S.","family":"So","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Andrew","family":"D'Arcangelo","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sarah","family":"Cancelarich","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Meghan C.","family":"Drummond","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Peter G.","family":"Slade","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ning","family":"Pan","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Tyler M.","family":"Gibson","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Tian","family":"Yang","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Joseph C.","family":"Burns","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Adam T.","family":"Palermo","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Lars","family":"Becker","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1965","published-online":{"date-parts":[[2024,6,17]]},"reference":[{"key":"ref1","doi-asserted-by":"publisher","first-page":"4496","DOI":"10.1073\/pnas.1817537116","article-title":"Dual AAV-mediated gene therapy restores hearing in a DFNB9 mouse model","volume":"116","author":"Akil","year":"2019","journal-title":"Proc. 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