{"database":"biostudies-literature","file_versions":[],"scores":{"citationCount":0,"reanalysisCount":0,"viewCount":271,"searchCount":0},"additional":{"submitter":["Ye L"],"funding":["NIDA NIH HHS","NCI NIH HHS","U.S. Department of Health &amp; Human Services | National Institutes of Health","NIH HHS","NIGMS NIH HHS"],"pagination":["1302-1313"],"full_dataset_link":["https://www.ebi.ac.uk/biostudies/studies/S-EPMC6834896"],"repository":["biostudies-literature"],"omics_type":["Unknown"],"volume":["37(11)"],"pubmed_abstract":["Targeting membrane proteins could improve the efficacy of T cell-based immunotherapies. To facilitate the identification of T cell targets, we developed a hybrid genetic screening system where the Sleeping Beauty (SB) transposon and single guide RNA cassette are nested in an adeno-associated virus (AAV). SB-mediated genomic integration of the single guide RNA cassette enables efficient gene editing in primary murine T cells as well as a screen readout. We performed in vivo AAV-SB-CRISPR screens for membrane protein targets in CD8<sup>+</sup> T cells in mouse models of glioblastoma (GBM). We validated screen hits by demonstrating that adoptive transfer of CD8<sup>+</sup> T cells with Pdia3, Mgat5, Emp1 or Lag3 gene editing enhances the survival of GBM-bearing mice in both syngeneic and T-cell receptor transgenic models. Transcriptome profiling, single cell sequencing, cytokine assays and T cell signaling analysis showed that Pdia3 editing in T cells enhances effector functions. Engineered PDIA3 mutant EGFRvIII chimeric antigen T cells are more potent in antigen-specific killing of human GBM cells."],"journal":["Nature biotechnology"],"pubmed_title":["In vivo CRISPR screening in CD8 T cells with AAV-Sleeping Beauty hybrid vectors identifies membrane targets for improving immunotherapy for glioblastoma."],"pmcid":["PMC6834896"],"funding_grant_id":["U54 CA209992","DP2 CA238295","RF1 DA048811","1R33CA225498","1U54CA209992-8697","T32 GM007205","S10 OD018521","R01 CA231112","1R01CA231112","R33 CA225498"],"pubmed_authors":["Ye L","Guo J","Park JJ","Errami Y","Chow RD","Du Y","Wang G","Yang Q","Peng L","Chen S","Dong MB","Dai X"],"view_count":["271"],"additional_accession":[]},"is_claimable":false,"name":"In vivo CRISPR screening in CD8 T cells with AAV-Sleeping Beauty hybrid vectors identifies membrane targets for improving immunotherapy for glioblastoma.","description":"Targeting membrane proteins could improve the efficacy of T cell-based immunotherapies. To facilitate the identification of T cell targets, we developed a hybrid genetic screening system where the Sleeping Beauty (SB) transposon and single guide RNA cassette are nested in an adeno-associated virus (AAV). SB-mediated genomic integration of the single guide RNA cassette enables efficient gene editing in primary murine T cells as well as a screen readout. We performed in vivo AAV-SB-CRISPR screens for membrane protein targets in CD8<sup>+</sup> T cells in mouse models of glioblastoma (GBM). We validated screen hits by demonstrating that adoptive transfer of CD8<sup>+</sup> T cells with Pdia3, Mgat5, Emp1 or Lag3 gene editing enhances the survival of GBM-bearing mice in both syngeneic and T-cell receptor transgenic models. Transcriptome profiling, single cell sequencing, cytokine assays and T cell signaling analysis showed that Pdia3 editing in T cells enhances effector functions. Engineered PDIA3 mutant EGFRvIII chimeric antigen T cells are more potent in antigen-specific killing of human GBM cells.","dates":{"release":"2019-01-01T00:00:00Z","publication":"2019 Nov","modification":"2024-11-12T00:05:12.442Z","creation":"2020-05-22T13:50:22Z"},"accession":"S-EPMC6834896","cross_references":{"pubmed":["31548728"],"doi":["10.1038/s41587-019-0246-4"]}}