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We have developed a synthetic biology strategy to generate HEK293\u2010based cell lines which have integrated essential AAV and adenoviral helper genes and are capable of producing rAAV upon induction. One such cell line, GX6B, produced up to 10<jats:sup>6<\/jats:sup> capsids per cell, but only a much lower level of rAAV genomes. The low AAV genome titer limited its rAAV productivity and increased empty viral particle content. To boost AAV genome amplification, the coding sequence of the DNA polymerase complex (UL30\/UL42) from helper Herpes Simplex Virus type 1 (HSV\u20101) was placed under an inducible promoter control and integrated into GX6B genome at a relatively low level. The resulting clones produced significantly higher titer of viral genomes, while their capsid level was unaffected. As a result, the encapsidated rAAV2 titer and the full particle content were significantly increased. We further demonstrated that this strategy of expressing HSV\u20101 DNA polymerase to increase full particle productivity could be implemented in a synthetic cell line producing another serotype rAAV8.<\/jats:p>","DOI":"10.1002\/bit.28885","type":"journal-article","created":{"date-parts":[[2024,11,23]],"date-time":"2024-11-23T01:05:37Z","timestamp":1732323937000},"page":"424-434","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":3,"title":["Expression of Viral DNA Polymerase in Synthetic Recombinant Adeno\u2010Associated Virus Producer Cell Line Enhances Full Particle Productivity"],"prefix":"10.1002","volume":"122","author":[{"given":"Yu\u2010Chieh","family":"Lin","sequence":"first","affiliation":[{"name":"Department of Chemical Engineering and Materials Science University of Minnesota Minneapolis Minnesota USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Han\u2010Jung","family":"Kuo","sequence":"additional","affiliation":[{"name":"Department of Chemical Engineering and Materials Science University of Minnesota Minneapolis Minnesota USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Min","family":"Lu","sequence":"additional","affiliation":[{"name":"Department of Chemical Engineering and Materials Science University of Minnesota Minneapolis Minnesota USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Carissa","family":"Rungkittikhun","sequence":"additional","affiliation":[{"name":"Department of Chemical Engineering and Materials Science University of Minnesota Minneapolis Minnesota USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7739-8692","authenticated-orcid":false,"given":"Wei\u2010Shou","family":"Hu","sequence":"additional","affiliation":[{"name":"Department of Chemical Engineering and Materials Science University of Minnesota Minneapolis Minnesota USA"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"311","published-online":{"date-parts":[[2024,11,22]]},"reference":[{"key":"e_1_2_9_2_1","doi-asserted-by":"publisher","DOI":"10.1371\/journal.ppat.1000340"},{"key":"e_1_2_9_3_1","doi-asserted-by":"publisher","DOI":"10.1111\/j.1600-0854.2006.00477.x"},{"key":"e_1_2_9_4_1","doi-asserted-by":"publisher","DOI":"10.2174\/156652310793797685"},{"key":"e_1_2_9_5_1","doi-asserted-by":"publisher","DOI":"10.1038\/sj.gt.3302611"},{"key":"e_1_2_9_6_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.virol.2005.06.017"},{"key":"e_1_2_9_7_1","doi-asserted-by":"publisher","DOI":"10.1038\/mtm.2016.2"},{"key":"e_1_2_9_8_1","doi-asserted-by":"publisher","DOI":"10.1089\/hum.2009.094"},{"key":"e_1_2_9_9_1","doi-asserted-by":"publisher","DOI":"10.1038\/s41467-021-26485-4"},{"key":"e_1_2_9_10_1","doi-asserted-by":"publisher","DOI":"10.1038\/s41598-021-01220-7"},{"key":"e_1_2_9_11_1","doi-asserted-by":"publisher","DOI":"10.1038\/mtm.2013.9"},{"key":"e_1_2_9_12_1","doi-asserted-by":"publisher","DOI":"10.1186\/1743-422X-2-43"},{"key":"e_1_2_9_13_1","doi-asserted-by":"publisher","DOI":"10.1038\/sj.gt.3300946"},{"key":"e_1_2_9_14_1","doi-asserted-by":"publisher","DOI":"10.1016\/S0021-9258(18)50411-7"},{"key":"e_1_2_9_15_1","doi-asserted-by":"publisher","DOI":"10.1007\/s00109-020-02034-2"},{"key":"e_1_2_9_16_1","doi-asserted-by":"publisher","DOI":"10.18609\/cgti.2018.016"},{"key":"e_1_2_9_17_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.ymthe.2022.12.001"},{"key":"e_1_2_9_18_1","doi-asserted-by":"publisher","DOI":"10.1128\/JVI.03428-12"},{"key":"e_1_2_9_19_1","doi-asserted-by":"publisher","DOI":"10.1093\/emboj\/20.12.3282"},{"key":"e_1_2_9_20_1","doi-asserted-by":"publisher","DOI":"10.1128\/jvi.00633-24"},{"key":"e_1_2_9_21_1","doi-asserted-by":"publisher","DOI":"10.1021\/acssynbio.2c00207"},{"key":"e_1_2_9_22_1","doi-asserted-by":"crossref","unstructured":"Lin Y. 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