{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,5,1]],"date-time":"2026-05-01T20:52:28Z","timestamp":1777668748822,"version":"3.51.4"},"reference-count":27,"publisher":"SAGE Publications","issue":"9-10","license":[{"start":{"date-parts":[[2026,2,19]],"date-time":"2026-02-19T00:00:00Z","timestamp":1771459200000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/journals.sagepub.com\/page\/policies\/text-and-data-mining-license"}],"content-domain":{"domain":["journals.sagepub.com"],"crossmark-restriction":true},"short-container-title":["Human Gene Therapy"],"published-print":{"date-parts":[[2026,5,1]]},"abstract":"\n Mutations in the\n Eyes Shut Homolog<\/jats:italic>\n (\n EYS<\/jats:italic>\n ) gene are one of the leading causes of autosomal recessive retinitis pigmentosa, a progressive retinal degenerative disease for which no effective treatment currently exists. However, the large size of the\n EYS<\/jats:italic>\n coding sequence (\u223c9.4 kb) exceeds the packaging limit of adeno-associated virus (AAV) vectors, posing a major barrier to gene replacement therapy. To address this challenge, we developed a tripartite AAV vector system that enables delivery and reconstitution of the full-length\n EYS<\/jats:italic>\n gene using a Cre-lox-based unidirectional DNA recombination strategy, Uni-directional and Site-specific Transgene Assembly by Recombination (Uni-STAR). The system consists of three AAV constructs carrying discrete\n EYS<\/jats:italic>\n segments flanked by engineered, noncompatible lox sites that drive ordered and unidirectional recombination in target cells. We validated this system\n in vitro<\/jats:italic>\n by demonstrating successful reconstitution and expression of full-length EYS protein in HEK293T cells.\n In vivo<\/jats:italic>\n , subretinal co-injection of the three AAV vectors into mouse eyes led to precise reconstitution and expression of full-length EYS protein in the retina. These findings establish the feasibility of using a tripartite AAV system to deliver the complete\n EYS<\/jats:italic>\n gene and provide a foundation for future therapeutic development targeting\n EYS<\/jats:italic>\n -associated retinal degenerations.\n <\/jats:p>","DOI":"10.1177\/10430342261426597","type":"journal-article","created":{"date-parts":[[2026,2,20]],"date-time":"2026-02-20T03:51:44Z","timestamp":1771559504000},"page":"442-452","update-policy":"https:\/\/doi.org\/10.1177\/sage-journals-update-policy","source":"Crossref","is-referenced-by-count":0,"title":["A Tripartite AAV System with Engineered Lox Sites Enables Efficient Delivery of the\n EYS<\/i>\n Gene for Retinal Gene Therapy"],"prefix":"10.1177","volume":"37","author":[{"given":"Kun-Do","family":"Rhee","sequence":"first","affiliation":[{"name":"Department of Ophthalmology and Visual Sciences, The University of Iowa Carver College of Medicine, Iowa City, Iowa, USA"},{"name":"Institute for Vision Research, The University of Iowa Carver College of Medicine, Iowa City, Iowa, USA."}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Poppy","family":"Datta","sequence":"additional","affiliation":[{"name":"Department of Ophthalmology and Visual Sciences, The University of Iowa Carver College of Medicine, Iowa City, Iowa, USA"},{"name":"Institute for Vision Research, The University of Iowa Carver College of Medicine, Iowa City, Iowa, USA."}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Clairissa","family":"Baccam","sequence":"additional","affiliation":[{"name":"Department of Ophthalmology and Visual Sciences, The University of Iowa Carver College of Medicine, Iowa City, Iowa, USA"},{"name":"Institute for Vision Research, The University of Iowa Carver College of Medicine, Iowa City, Iowa, USA."}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-7908-3210","authenticated-orcid":false,"given":"Seongjin","family":"Seo","sequence":"additional","affiliation":[{"name":"Department of Ophthalmology and Visual Sciences, The University of Iowa Carver College of Medicine, Iowa City, Iowa, USA"},{"name":"Institute for Vision Research, The University of Iowa Carver College of Medicine, Iowa City, Iowa, 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