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Cas9‐Cleavage Sequences in Size‐Reduced Plasmids Enhance Nonviral Genome Targeting of CARs in Primary Human T Cells
Author(s) -
Jing Ruirui,
Jiao Peng,
Chen Jiangqing,
Meng Xianhui,
Wu Xiaoyan,
Duan Yanting,
Shang Kai,
Qian Liling,
Huang Yanjie,
Liu Junwei,
Huang Tao,
Jin Jin,
Chen Wei,
Zeng Xun,
Yin Weiwei,
Gao Xiaofei,
Zhou Chun,
Sadelain Michel,
Sun Jie
Publication year - 2021
Publication title -
small methods
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 4.66
H-Index - 46
ISSN - 2366-9608
DOI - 10.1002/smtd.202100071
Subject(s) - chimeric antigen receptor , plasmid , genome editing , homologous recombination , biology , genome engineering , genome , crispr , cas9 , viral vector , dna , vector (molecular biology) , computational biology , microbiology and biotechnology , virology , genetics , recombinant dna , gene , t cell , immune system
T cell genome editing holds great promise to advance a range of immunotherapies but is encumbered by the dependence on difficult‐to‐produce and expensive viral vectors. Here, small double‐stranded plasmid DNA modified to mediate high‐efficiency homologous recombination is designed. The resulting chimeric antigen receptor (CAR)‐T cells display a similar phenotype, transcriptional profile, and in vivo potency to CAR‐T cells generated using adeno‐associated viral vector. This method should simplify and accelerate the use of precision engineering to produce edited T cells for research and clinical purposes.