{"database":"biostudies-literature","file_versions":[],"scores":null,"additional":{"submitter":["Tieu V"],"funding":["NCI NIH HHS","NIH","CIRM","National Science Foundation"],"pagination":["1278-1295.e20"],"full_dataset_link":["https://www.ebi.ac.uk/biostudies/studies/S-EPMC10965243"],"repository":["biostudies-literature"],"omics_type":["Unknown"],"volume":["187(5)"],"pubmed_abstract":["CRISPR technologies have begun to revolutionize T cell therapies; however, conventional CRISPR-Cas9 genome-editing tools are limited in their safety, efficacy, and scope. To address these challenges, we developed multiplexed effector guide arrays (MEGA), a platform for programmable and scalable regulation of the T cell transcriptome using the RNA-guided, RNA-targeting activity of CRISPR-Cas13d. MEGA enables quantitative, reversible, and massively multiplexed gene knockdown in primary human T cells without targeting or cutting genomic DNA. Applying MEGA to a model of CAR T cell exhaustion, we robustly suppressed inhibitory receptor upregulation and uncovered paired regulators of T cell function through combinatorial CRISPR screening. We additionally implemented druggable regulation of MEGA to control CAR activation in a receptor-independent manner. Lastly, MEGA enabled multiplexed disruption of immunoregulatory metabolic pathways to enhance CAR T cell fitness and anti-tumor activity in vitro and in vivo. MEGA offers a versatile synthetic toolkit for applications in cancer immunotherapy and beyond."],"journal":["Cell"],"pubmed_title":["A versatile CRISPR-Cas13d platform for multiplexed transcriptomic regulation and metabolic engineering in primary human T cells."],"pmcid":["PMC10965243"],"funding_grant_id":["R21 CA270609","P30 CA124435"],"pubmed_authors":["Quinn PJ","Guerrero JA","Xu P","Tieu V","Sotillo E","Malipatlolla M","Mackall CL","Qi LS","Chen C","Bjelajac JR","Klysz D","Fisher C"],"additional_accession":[]},"is_claimable":false,"name":"A versatile CRISPR-Cas13d platform for multiplexed transcriptomic regulation and metabolic engineering in primary human T cells.","description":"CRISPR technologies have begun to revolutionize T cell therapies; however, conventional CRISPR-Cas9 genome-editing tools are limited in their safety, efficacy, and scope. To address these challenges, we developed multiplexed effector guide arrays (MEGA), a platform for programmable and scalable regulation of the T cell transcriptome using the RNA-guided, RNA-targeting activity of CRISPR-Cas13d. MEGA enables quantitative, reversible, and massively multiplexed gene knockdown in primary human T cells without targeting or cutting genomic DNA. Applying MEGA to a model of CAR T cell exhaustion, we robustly suppressed inhibitory receptor upregulation and uncovered paired regulators of T cell function through combinatorial CRISPR screening. We additionally implemented druggable regulation of MEGA to control CAR activation in a receptor-independent manner. Lastly, MEGA enabled multiplexed disruption of immunoregulatory metabolic pathways to enhance CAR T cell fitness and anti-tumor activity in vitro and in vivo. MEGA offers a versatile synthetic toolkit for applications in cancer immunotherapy and beyond.","dates":{"release":"2024-01-01T00:00:00Z","publication":"2024 Feb","modification":"2026-06-02T20:08:33.913Z","creation":"2025-04-07T07:53:32.392Z"},"accession":"S-EPMC10965243","cross_references":{"pubmed":["38387457"],"doi":["10.1016/j.cell.2024.01.035"]}}