{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,16]],"date-time":"2026-10-16T07:10:36Z","timestamp":1792134636939,"version":"4.4.0"},"reference-count":38,"publisher":"Wiley","issue":"3","license":[{"start":{"date-parts":[[2025,1,23]],"date-time":"2025-01-23T00:00:00Z","timestamp":1737590400000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by-nc\/4.0\/"}],"content-domain":{"domain":["aiche.onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["Biotechnology Progress"],"published-print":{"date-parts":[[2025,5]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                  <jats:p>\n                    We present the first use of a bioengineered mammalian transposase system derived from\n                    <jats:italic>Myotis lucifugus<\/jats:italic>\n                    (bMLT) for integration of expression vectors into the CHO genome, focusing on GFP and trastuzumab production. Initially, CHO\u2010K1 cells are transfected with a GFP reporter and varying amounts of bMLT DNA or mRNA. GFP expression is monitored over 17\u2009weeks without selective pressure. Transfection efficiency shows around 90% GFP\u2010positive cells, but in control cultures GFP expression disappears after 10\u2009days. In contrast, bMLT\u2010treated cultures maintain stable GFP expression, with a dose\u2010dependent integration efficiency of up to 60%. The highest GFP expression per cell is observed with lower bMLT amounts. Next\u2010generation sequencing analysis reveals multiple integration sites, with 85% correctly integrated sequences. Next, CHO\u2010GS\n                    <jats:sup>\u2212\/\u2212<\/jats:sup>\n                    cells are transfected with trastuzumab and bMLT DNA or mRNA. Cells are selected in glutamine\u2010free medium with varying methionine sulfoximine (MSX) concentrations. Recovery is faster without MSX, and no difference is observed between bMLT DNA and mRNA transfections. bMLT\u2010treated cultures show a higher percentage of trastuzumab\u2010secreting cells (40%\u201355%) compared with random integration (0.3%\u20130.5%). The absence of insulators in the trastuzumab plasmid likely affects selection behavior, as integration in heterochromatic regions results in gene repression. Overall, bMLT\u2010mediated integration proves efficient, generating stable cell pools with high expression profiles without selective pressure. The integration sites' genomic location significantly impacts productivity, with favorable regions supporting higher expression. This method shows promise for the rapid and efficient generation of high\u2010producing cell lines and for rapid evaluation of long\u2010term effects of different cell engineering approaches.\n                  <\/jats:p>","DOI":"10.1002\/btpr.3524","type":"journal-article","created":{"date-parts":[[2025,1,23]],"date-time":"2025-01-23T08:31:36Z","timestamp":1737621096000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":1,"title":["Characterization of large transgene integrations in Chinese hamster ovary cells using a bioengineered mammalian transposase"],"prefix":"10.1002","volume":"41","author":[{"given":"Nicolas","family":"Marx","sequence":"first","affiliation":[{"name":"Department of Biotechnology BOKU University of Natural Resources and Life Sciences  Vienna Austria"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Anna\u2010Maria","family":"Otte","sequence":"additional","affiliation":[{"name":"Department of Biotechnology BOKU University of Natural Resources and Life Sciences  Vienna Austria"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Klaus","family":"Leitner","sequence":"additional","affiliation":[{"name":"Department of Biotechnology BOKU University of Natural Resources and Life Sciences  Vienna Austria"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Rehmadanta","family":"Sitepu","sequence":"additional","affiliation":[{"name":"Department of Biotechnology BOKU University of Natural Resources and Life Sciences  Vienna Austria"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Thomas","family":"Berger","sequence":"additional","affiliation":[{"name":"Bioanalytical Research Labs Paris\u2010Lodron University Salzburg  Salzburg Austria"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Veronika","family":"Sch\u00e4pert\u00f6ns","sequence":"additional","affiliation":[{"name":"Bioanalytical Research Labs Paris\u2010Lodron University Salzburg  Salzburg Austria"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Christian G.","family":"Huber","sequence":"additional","affiliation":[{"name":"Bioanalytical Research Labs Paris\u2010Lodron University Salzburg  Salzburg Austria"},{"name":"Center for Tumor Biology and Immunology Paris\u2010Lodron University Salzburg  Salzburg Austria"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Quan","family":"Zhu","sequence":"additional","affiliation":[{"name":"SalioGen Therapeutics  Lexington Massachusetts USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sandeep","family":"Nema","sequence":"additional","affiliation":[{"name":"SalioGen Therapeutics  Lexington Massachusetts USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Joseph J.","family":"Higgins","sequence":"additional","affiliation":[{"name":"SalioGen Therapeutics  Lexington Massachusetts USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6324-9338","authenticated-orcid":false,"given":"Nicole","family":"Borth","sequence":"additional","affiliation":[{"name":"Department of Biotechnology BOKU University of Natural Resources and Life Sciences  Vienna 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