Cas9‐Cleavage Sequences in Size‐Reduced Plasmids Enhance Nonviral Genome Targeting of CARs in Primary Human T Cells
Ruirui Jing(Zhejiang Chinese Medical University), Peng Jiao(Zhejiang Chinese Medical University), Jiangqing Chen(Zhejiang Chinese Medical University), Xianhui Meng(Zhejiang Chinese Medical University), Xiaoyan Wu(Zhejiang Chinese Medical University), Yanting Duan(Zhejiang Chinese Medical University), Kai Shang(Zhejiang Chinese Medical University), Liling Qian(First Affiliated Hospital Zhejiang University), Yanjie Huang(Westlake University), Junwei Liu(Ministry of Education), Tao Huang(Zhejiang University), Jin Jin(Zhejiang University), Wei Chen(Second Affiliated Hospital of Zhejiang University), Xun Zeng(First Affiliated Hospital Zhejiang University), Weiwei Yin(Ministry of Education), Xiaofei Gao(Westlake University), Chun Zhou(Sir Run Run Shaw Hospital), Michel Sadelain(Kettering University), Jie Sun(Zhejiang Chinese Medical University)
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Abstract
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.
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