{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,7]],"date-time":"2026-10-07T04:17:13Z","timestamp":1791346633342,"version":"4.3.1"},"reference-count":82,"publisher":"American Association for the Advancement of Science (AAAS)","issue":"824","funder":[{"name":"CFF","award":["CFF HE21I0"],"award-info":[{"award-number":["CFF HE21I0"]}]},{"name":"NIH NIGMS","award":["NIH 1R35GM133794"],"award-info":[{"award-number":["NIH 1R35GM133794"]}]},{"DOI":"10.13039\/100001797","name":"PhRMA Foundation","doi-asserted-by":"crossref","award":["100001797"],"award-info":[{"award-number":["100001797"]}],"id":[{"id":"10.13039\/100001797","id-type":"DOI","asserted-by":"crossref"}]},{"name":"University of Florida Health Cancer Center GU pilot"}],"content-domain":{"domain":["www.science.org"],"crossmark-restriction":true},"short-container-title":["Sci. Transl. Med."],"published-print":{"date-parts":[[2025,11,12]]},"abstract":"<jats:p>\n                    The clinical translation of gene therapy has been challenging in part because of the limitations of current delivery approaches. Herein, we report an efficient nonviral genome editor delivery approach using extracellular vesicles (EVs) carrying single-guide RNA (sgRNA): CRISPR-Cas9 ribonucleoprotein (RNP) complexes for in vivo gene therapy. By leveraging a high-throughput microfluidic droplet\u2013based electroporation system (\u03bcDES), we achieved a 10-fold enhancement in loading efficiency and more than 1000-fold increase in processing throughput for loading RNP complexes into EVs compared with conventional high-voltage pulsed electroporation. \u03bcDES generated uniform microdroplets containing EVs and RNPs by applying direct current\u2013controlled low voltage (up to 60 V) to transiently permeabilize membranes and enable efficient cargo encapsulation while maintaining EV integrity at both the protein and morphological levels. In the\n                    <jats:italic toggle=\"yes\">\n                      Myo7a\n                      <jats:sup>WT\/Sh1<\/jats:sup>\n                    <\/jats:italic>\n                    mouse model of autosomal dominant progressive hearing loss, which may model MYO7A-associated DFNA11 hearing loss in humans, we demonstrated the effective delivery of RNPs by EVs into cochlear hair cells by cross-sectional and whole-mount confocal imaging. The injection of RNP-EVs via the posterior semicircular canal in 4-week-old\n                    <jats:italic toggle=\"yes\">\n                      Myo7a\n                      <jats:sup>WT\/Sh1<\/jats:sup>\n                    <\/jats:italic>\n                    mice resulted in a reduction in\n                    <jats:italic toggle=\"yes\">\n                      Myo7a\n                      <jats:sup>Sh1<\/jats:sup>\n                    <\/jats:italic>\n                    messenger RNA expression and evidence of hearing preservation, as measured by auditory brainstem responses, compared with untreated ears and EV only\u2013injected mice. This study highlights the potential of \u03bcDES-produced RNP-EVs for gene editing as a treatment for progressive nonsyndromic hearing loss in patients.\n                  <\/jats:p>","DOI":"10.1126\/scitranslmed.adn3993","type":"journal-article","created":{"date-parts":[[2025,11,12]],"date-time":"2025-11-12T18:58:19Z","timestamp":1762973899000},"update-policy":"https:\/\/doi.org\/10.34133\/aaas_crossmark","source":"Crossref","is-referenced-by-count":9,"title":["Extracellular vesicle\u2013mediated gene editing for the treatment of nonsyndromic progressive hearing loss in adult mice"],"prefix":"10.1126","volume":"17","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-1896-2198","authenticated-orcid":true,"given":"Xiaoshu","family":"Pan","sequence":"first","affiliation":[{"name":"College of Pharmacy, Department of Pharmaceutics, University of Florida, Gainesville, FL 32611, 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