{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,1,8]],"date-time":"2026-01-08T21:34:19Z","timestamp":1767908059571,"version":"3.49.0"},"reference-count":51,"publisher":"MDPI AG","issue":"14","license":[{"start":{"date-parts":[[2022,7,7]],"date-time":"2022-07-07T00:00:00Z","timestamp":1657152000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"ReiThera s.r.l"},{"name":"2017 MILLION DOLLAR BIKE RIDE Pilot"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>Sleeping Beauty (SB) is the first DNA transposon employed for efficient transposition in vertebrate cells, opening new applications for genetic engineering and gene therapies. A transposon-based gene delivery system holds the favourable features of non-viral vectors and an attractive safety profile. Here, we employed SB to engineer HEK293 cells for optimizing the production of a chimpanzee Adenovector (chAd) belonging to the Human Mastadenovirus C species. To date, chAd vectors are employed in several clinical settings for infectious diseases, last but not least COVID-19. A robust, efficient and quick viral vector production could advance the clinical application of chAd vectors. To this aim, we firstly swapped the hAd5 E1 with chAd-C E1 gene by using the CRISPR\/Cas9 system. We demonstrated that in the absence of human Ad5 E1, chimp Ad-C E1 gene did not support HEK293 survival. To improve chAd-C vector production, we engineered HEK293 cells to stably express the chAd-C precursor terminal protein (ch.pTP), which plays a crucial role in chimpanzee Adenoviral DNA replication. The results indicate that exogenous ch.pTP expression significantly ameliorate the packaging and amplification of recombinant chAd-C vectors thus, the engineered HEK293ch.pTP cells could represent a superior packaging cell line for the production of these vectors.<\/jats:p>","DOI":"10.3390\/ijms23147538","type":"journal-article","created":{"date-parts":[[2022,7,7]],"date-time":"2022-07-07T22:11:47Z","timestamp":1657231907000},"page":"7538","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":3,"title":["Engineered Sleeping Beauty Transposon as Efficient System to Optimize Chimp Adenoviral Production"],"prefix":"10.3390","volume":"23","author":[{"given":"Samantha","family":"Baldassarri","sequence":"first","affiliation":[{"name":"Centre for Regenerative Medicine, Department of Life Sciences, University of Modena and Reggio Emilia, 41121 Modena, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6599-7193","authenticated-orcid":false,"given":"Daniela","family":"Benati","sequence":"additional","affiliation":[{"name":"Centre for Regenerative Medicine, Department of Life Sciences, University of Modena and Reggio Emilia, 41121 Modena, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Federica","family":"D\u2019Alessio","sequence":"additional","affiliation":[{"name":"ReiThera S.r.l., 00128 Rome, Italy"},{"name":"Department of Molecular Medicine and Medical Biotechnologies, University of Naples \u201cFederico II\u201d, 80138 Naples, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Clarissa","family":"Patrizi","sequence":"additional","affiliation":[{"name":"Centre for Regenerative Medicine, Department of Life Sciences, University of Modena and Reggio Emilia, 41121 Modena, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Eleonora","family":"Cattin","sequence":"additional","affiliation":[{"name":"Centre for Regenerative Medicine, Department of Life Sciences, University of Modena and Reggio Emilia, 41121 Modena, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Michela","family":"Gentile","sequence":"additional","affiliation":[{"name":"ReiThera S.r.l., 00128 Rome, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6748-0307","authenticated-orcid":false,"given":"Angelo","family":"Raggioli","sequence":"additional","affiliation":[{"name":"ReiThera S.r.l., 00128 Rome, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Alessandra","family":"Recchia","sequence":"additional","affiliation":[{"name":"Centre for Regenerative Medicine, Department of Life Sciences, University of Modena and Reggio Emilia, 41121 Modena, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2022,7,7]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"2083","DOI":"10.1056\/NEJMoa1310566","article-title":"Efficacy trial of a DNA\/rAd5 HIV-1 preventive vaccine","volume":"369","author":"Hammer","year":"2013","journal-title":"N. 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