Journal
SMALL METHODS
Volume 5, Issue 7, Pages -Publisher
WILEY-V C H VERLAG GMBH
DOI: 10.1002/smtd.202100071
Keywords
adoptive cell therapy; CAR; CRISPR/Cas9; genome targeting; nonviral methods; size-reduced plasmid
Funding
- Zhejiang Provincial Key Laboratory of Immunity and Inflammatory diseases - National Natural Science Foundation of China [31971324, 31971125]
- Zhejiang Provincial Natural Science Foundation [LR20H160003]
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Genome editing of T cells using modified plasmid DNA can simplify and expedite the process of producing CAR-T cells, increasing their potency for research and clinical purposes.
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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