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Currently, rAAV is commonly produced by plasmid transient transfection (TT)\u00a0into host cells; however, the requirement for transfection limits scalability in manufacturing. To overcome this limitation, transfection-free production methods using inducible producer cell lines (PCLs)\u2013in which essential genes for rAAV production are integrated into the host genome\u2013have been actively developed. Despite the promise of PCL technology, comprehensive evidence showing that PCL-derived rAAV particles are equivalent to those produced by the conventional TT method has been lacking. In this study, we established an inducible rAAV PCL using a newly designed single plasmid harboring all genes necessary for rAAV production. Through a single selection step, we successfully isolated a high-yield single-cell clone. Physicochemical analyses of the produced rAAV, including capsid protein composition, genome packaging length, and sedimentation coefficient, confirmed equivalence between PCL-derived rAAV and that produced by conventional transient transfection. Furthermore, biological evaluation demonstrated that PCL-derived and transfection-derived rAAV exhibited comparable infectivity and transduction profiles both in HeLa cells in vitro and in the mouse brain in vivo. These results demonstrate that PCL-derived rAAV particles are equivalent to those from the TT method, highlighting that our inducible PCL approach is a promising platform for scalable rAAV production.<\/jats:p>","DOI":"10.1038\/s41598-026-56897-5","type":"journal-article","created":{"date-parts":[[2026,6,26]],"date-time":"2026-06-26T10:14:28Z","timestamp":1782468868000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Inducible rAAV producer cell lines yield vectors equivalent to transient transfection: a physicochemical and biological comparison"],"prefix":"10.1038","volume":"16","author":[{"given":"Yuki","family":"Uchida","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yusuke","family":"Mori","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Takaaki","family":"Kurinomaru","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ryo","family":"Aoki","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Susumu","family":"Uchiyama","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hirokazu","family":"Hirai","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Taichi","family":"Muraguchi","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"297","published-online":{"date-parts":[[2026,6,26]]},"reference":[{"key":"56897_CR1","doi-asserted-by":"publisher","first-page":"78","DOI":"10.1038\/s41392-024-01780-w","volume":"9","author":"JH Wang","year":"2024","unstructured":"Wang, J. 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