{"database":"biostudies-literature","file_versions":[],"scores":null,"additional":{"submitter":["Webber BR"],"funding":["Employee of Intima Bioscience","Sponsored Research Agreement funded by Intima Biosciences","NIAID NIH HHS","NHLBI NIH HHS","Foundation for the National Institutes of Health","Foundation for the National Institutes of Health (Foundation for the National Institutes of Health, Inc.)","NCI NIH HHS"],"pagination":["1553-1570"],"full_dataset_link":["https://www.ebi.ac.uk/biostudies/studies/S-EPMC11169092"],"repository":["biostudies-literature"],"omics_type":["Unknown"],"volume":["8(12)"],"pubmed_abstract":["The reliance on viral vectors for the production of genetically engineered immune cells for adoptive cellular therapies remains a translational bottleneck. Here we report a method leveraging the DNA repair pathway homology-mediated end joining, as well as optimized reagent composition and delivery, for the Cas9-induced targeted integration of large DNA payloads into primary human T cells with low toxicity and at efficiencies nearing those of viral vectors (targeted knock-in of 1-6.7 kb payloads at rates of up to 70% at multiple targeted genomic loci and with cell viabilities of over 80%). We used the method to produce T cells with an engineered T-cell receptor or a chimaeric antigen receptor and show that the cells maintained low levels of exhaustion markers and excellent capacities for pr"],"journal":["Nature biomedical engineering"],"pubmed_title":["Cas9-induced targeted integration of large DNA payloads in primary human T cells via homology-mediated end-joining DNA repair."],"pmcid":["PMC11169092"],"funding_grant_id":["R01 AI146009","T32 HL007062","P01 CA254849","R37 CA276345","R01 AI161017","T32HL007062-46"],"pubmed_authors":["Skeate JG","Starr TK","Henley T","Diers MD","McIvor RS","Slipek NJ","Webber BR","Johnson MJ","Lahr WS","Moriarity BS","Qiu X","Rathmann B","Choudhry M","Wick B","DeFeo AP","Mills LJ"],"additional_accession":[]},"is_claimable":false,"name":"Cas9-induced targeted integration of large DNA payloads in primary human T cells via homology-mediated end-joining DNA repair.","description":"The reliance on viral vectors for the production of genetically engineered immune cells for adoptive cellular therapies remains a translational bottleneck. Here we report a method leveraging the DNA repair pathway homology-mediated end joining, as well as optimized reagent composition and delivery, for the Cas9-induced targeted integration of large DNA payloads into primary human T cells with low toxicity and at efficiencies nearing those of viral vectors (targeted knock-in of 1-6.7 kb payloads at rates of up to 70% at multiple targeted genomic loci and with cell viabilities of over 80%). We used the method to produce T cells with an engineered T-cell receptor or a chimaeric antigen receptor and show that the cells maintained low levels of exhaustion markers and excellent capacities for pr","dates":{"release":"2024-01-01T00:00:00Z","publication":"2024 Dec","modification":"2026-06-02T18:12:46.702Z","creation":"2025-04-06T17:33:34.857Z"},"accession":"S-EPMC11169092","cross_references":{"pubmed":["38092857"],"doi":["10.1038/s41551-023-01157-4"]}}