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In this study, we employed the programmable nuclease (PN) Cas\u2010CLOVER to precisely target the Glutamine synthetase (GS) locus in CHO cells. A total of 30 unique serum\u2010free, suspension\u2010adapted CHO\u2010K1 candidate host cell lines were subjected to Cas\u2010CLOVER\u2010mediated gene editing, generating over one hundred potential GS knockout (GSKO) clones. A subset of the GSKO clones was subsequently validated using three orthogonal methods to confirm complete knockout of the GS gene in 98 clones. Randomly selected GSKO clones were utilized to produce standard monoclonal antibodies. The resulting pools demonstrated enhanced productivity, with up to a 14.5\u2010fold increase in titer compared to their wild\u2010type parental hosts. These findings highlight the potential of gene editing approaches to significantly improve recombinant protein production in CHO expression systems, offering valuable insights for biopharmaceutical manufacturing applications.<\/jats:p>","DOI":"10.1002\/btpr.70108","type":"journal-article","created":{"date-parts":[[2026,2,9]],"date-time":"2026-02-09T03:47:24Z","timestamp":1770608844000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Evaluation of gene editing in\n                    <scp>CHO<\/scp>\n                    cells using the Cas\u2010\n                    <scp>CLOVER<\/scp>\n                    system"],"prefix":"10.1002","volume":"42","author":[{"ORCID":"https:\/\/orcid.org\/0009-0005-1294-0731","authenticated-orcid":false,"given":"Tiffany","family":"McLamarrah","sequence":"first","affiliation":[{"name":"Mammalian Platform, Global CMC Development  Framingham Massachusetts 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