{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,1]],"date-time":"2026-10-01T16:56:53Z","timestamp":1790873813998,"version":"4.1.0"},"reference-count":37,"publisher":"Frontiers Media SA","license":[{"start":{"date-parts":[[2026,7,27]],"date-time":"2026-07-27T00:00:00Z","timestamp":1785110400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":["frontiersin.org"],"crossmark-restriction":true},"short-container-title":["Front. Immunol."],"abstract":"<jats:sec>\n                    <jats:title>Introduction<\/jats:title>\n                    <jats:p>CRISPR\u2013Cas9 has transformed the engineering of chimeric antigen receptor T (CAR-T) cells and chimeric antigen receptor NK (CAR-NK) cells; however, its clinical translation remains constrained by the high cost, batch-to-batch variability, and stringent regulatory requirements associated with current viral and electroporation-based manufacturing approaches.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Methods<\/jats:title>\n                    <jats:p>We report an industrial-grade platform based on monoclonal producer cell lines that enables the continuous and scalable generation of engineered virus-like particles (eVLPs) co-packaging Cas9\u2013gRNA ribonucleoproteins (RNPs). A progenitor cell line was established by stably integrating three core modules\u2014Gag-Pol, Gag-Cas9, and the baboon endogenous virus (BaEV) envelope\u2014into a single HEK293T clone. Introduction of a self-inactivating (SIN) retroviral vector encoding the gRNA cassette (exemplified here by CD7) converted this progenitor into a dedicated eVLP producer within 10 days.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Results<\/jats:title>\n                    <jats:p>Using this platform, we generated CD7-knockout CAR-T\/NK cells that retained robust in vitro cytotoxicity, confirming preserved functional activity. Owing to its modular architecture, the platform is readily extensible. For example, integration with Recombinant Adeno-associated Virus (rAAV) donor templates enables site-specific CAR insertion, while multiplexed eVLP cocktails allow simultaneous disruption of multiple genomic loci.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Discussion<\/jats:title>\n                    <jats:p>It is worth noting that this workflow eliminates the need for electroporation, reduces serum dependency, and significantly lowers the cost of reagent consumables. Collectively, this system provides a GMP-compliant and broadly adaptable strategy for the streamlined manufacturing of next-generation autologous and allogeneic gene-edited CAR-T\/NK therapies.<\/jats:p>\n                  <\/jats:sec>","DOI":"10.3389\/fimmu.2026.1878099","type":"journal-article","created":{"date-parts":[[2026,7,27]],"date-time":"2026-07-27T05:40:34Z","timestamp":1785130834000},"update-policy":"https:\/\/doi.org\/10.3389\/crossmark-policy","source":"Crossref","is-referenced-by-count":1,"title":["Establishment of a scalable engineered cell-line platform for direct, GMP-grade production of eVLP vectors enabling streamlined generation of gene-edited CAR-T\/NK cells"],"prefix":"10.3389","volume":"17","author":[{"given":"Wei","family":"Lin","sequence":"first","affiliation":[{"name":"School of Life Sciences, Beijing University of Chinese Medicine","place":["Beijing, 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China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Lili","family":"Chen","sequence":"additional","affiliation":[{"name":"Guangdong Junhou Biopharmaceuticals Co., LTD","place":["Zhongshan, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Shengshui","family":"Mo","sequence":"additional","affiliation":[{"name":"Guangdong Junhou Biopharmaceuticals Co., LTD","place":["Zhongshan, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Zhengbo","family":"Wang","sequence":"additional","affiliation":[{"name":"Shenzhen Cell Valley Biopharmaceuticals Co., LTD","place":["Shenzhen, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Huaxiu","family":"Li","sequence":"additional","affiliation":[{"name":"Shenzhen Cell Valley Biopharmaceuticals Co., LTD","place":["Shenzhen, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yaru","family":"Feng","sequence":"additional","affiliation":[{"name":"Guangdong Junhou Biopharmaceuticals Co., LTD","place":["Zhongshan, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Lijun","family":"Zhao","sequence":"additional","affiliation":[{"name":"Shenzhen Cell Valley Biopharmaceuticals Co., LTD","place":["Shenzhen, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Junhui","family":"Chen","sequence":"additional","affiliation":[{"name":"Department of Minimally Invasion Intervention, Peking University Shenzhen Hospital","place":["Shenzhen, China"]},{"name":"Intervention and Cell Therapy Center, Peking University Shenzhen Hospital","place":["Shenzhen, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Guohua","family":"Yu","sequence":"additional","affiliation":[{"name":"School of Life Sciences, Beijing University of Chinese Medicine","place":["Beijing, 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