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Here, we demonstrate that CRISPR\u2010 and shRNA\u2010based cell engineering strategies can be used to disrupt RVLP production by targeting the RVLP nucleotide sequences. Additionally, specific antibodies were generated to monitor RVLP protein expression, including RVLP envelope (Env) protein localized on the surface of CHO cells, greatly facilitating selection of RVLP\u2010deficient clones. These modified CHO cells showed reduced RVLP production while maintaining or enhancing the ability to produce recombinant virus\u2010like particles (VLPs), highlighting their potential application in biomanufacturing, especially for complex biologics that are incompatible with standard RVLP mitigation procedures, namely viral inactivation and nanofiltration.<\/jats:p>","DOI":"10.1002\/bit.70043","type":"journal-article","created":{"date-parts":[[2025,8,22]],"date-time":"2025-08-22T10:30:41Z","timestamp":1755858641000},"page":"3192-3204","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Endogenous Retrovirus\u2010Like Particle\u2010Deficient CHO Cells Can be Generated by CRISPR or shRNA and Enriched Based on Cell\u2010Surface Expression of Retroviral Envelope Protein"],"prefix":"10.1002","volume":"122","author":[{"ORCID":"https:\/\/orcid.org\/0009-0002-4403-0677","authenticated-orcid":false,"given":"Matthew","family":"Stuible","sequence":"first","affiliation":[{"name":"National Research Council Canada, Human Health Therapeutics Research Centre Montreal Quebec Canada"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5958-2880","authenticated-orcid":false,"given":"Sergio P.","family":"Alpuche\u2010Lazcano","sequence":"additional","affiliation":[{"name":"National Research Council Canada, Human Health Therapeutics Research Centre Montreal Quebec Canada"},{"name":"Current affiliation: Departamento de Medicina Molecular y Bioprocesos, Instituto de Biotecnolog\u00eda Universidad Nacional Aut\u00f3noma de M\u00e9xico (UNAM) Cuernavaca Morelos CP 62210 M\u00e9xico"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Christian","family":"Gervais","sequence":"additional","affiliation":[{"name":"National Research Council Canada, Human Health Therapeutics Research Centre Montreal Quebec Canada"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Manon","family":"Ouimet","sequence":"additional","affiliation":[{"name":"National Research Council Canada, Human Health Therapeutics Research Centre Montreal Quebec Canada"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Julie","family":"Lippens","sequence":"additional","affiliation":[{"name":"National Research Council Canada, Human Health Therapeutics Research Centre Montreal Quebec Canada"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Martine","family":"Pag\u00e9","sequence":"additional","affiliation":[{"name":"National Research Council Canada, Human Health Therapeutics Research Centre Montreal Quebec Canada"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Audrey","family":"Morasse","sequence":"additional","affiliation":[{"name":"National Research Council Canada, Human Health Therapeutics Research Centre Montreal Quebec Canada"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3416-480X","authenticated-orcid":false,"given":"Anna N.","family":"Moraitis","sequence":"additional","affiliation":[{"name":"National Research Council Canada, Human Health Therapeutics Research Centre Montreal Quebec Canada"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2268-4111","authenticated-orcid":false,"given":"Yves","family":"Durocher","sequence":"additional","affiliation":[{"name":"National Research Council Canada, Human Health Therapeutics Research Centre Montreal Quebec Canada"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"311","published-online":{"date-parts":[[2025,8,22]]},"reference":[{"key":"e_1_2_9_2_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.crbiot.2022.03.002"},{"key":"e_1_2_9_3_1","doi-asserted-by":"publisher","DOI":"10.1038\/s43856-023-00340-7"},{"key":"e_1_2_9_4_1","doi-asserted-by":"publisher","DOI":"10.1016\/0042-6822(91)90496-X"},{"key":"e_1_2_9_5_1","doi-asserted-by":"publisher","DOI":"10.1002\/bit.28578"},{"key":"e_1_2_9_6_1","doi-asserted-by":"publisher","DOI":"10.1161\/ATVBAHA.108.179655"},{"key":"e_1_2_9_7_1","doi-asserted-by":"publisher","DOI":"10.1128\/jvi.02496-06"},{"key":"e_1_2_9_8_1","doi-asserted-by":"publisher","DOI":"10.1128\/jvi.74.8.3709-3714.2000"},{"key":"e_1_2_9_9_1","doi-asserted-by":"publisher","DOI":"10.1002\/bit.27200"},{"key":"e_1_2_9_10_1","unstructured":"Fire A. 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