Cas9-cleavage sequences in minimal plasmids enhance non-viral genome targeting of CARs in primary human T cells

Jing, R.; Jiao, P.; chen, j.; Meng, X.; Wu, X.; Duan, Y.; Shang, K.; Qian, L.; Huang, Y.; Liu, J.; Huang, T.; Jin, J.; Chen, W.; Zeng, X.; Yin, W.; Gao, X.; zhou, c.; Sedelain, M.; Sun, J.

doi:10.1101/2020.12.31.424920

bioRxiv · 10.1101/2020.12.31.424920

Cas9-cleavage sequences in minimal plasmids enhance non-viral genome targeting of CARs in primary human T cells

Jing, R.·Jiao, P.·chen, j.·Meng, X.·Wu, X.·Duan, Y.·Shang, K.·Qian, L.·Huang, Y.·Liu, J.·Huang, T.·Jin, J.·Chen, W.·Zeng, X.·Yin, W.·Gao, X.·zhou, c.·Sedelain, M.·Sun, J.

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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 we have designed small double-stranded plasmid DNA modified to mediate high-efficiency homologous recombination. The resulting chimeric antigen receptor (CAR)-T cells display a similar phenotype, transcriptional profile and in vivo potency as CAR-T cells generated using adeno-associated viral (AAV) 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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Jing, R., Jiao, P., chen, j., Meng, X., Wu, X., Duan, Y., Shang, K., Qian, L., Huang, Y., Liu, J., Huang, T., Jin, J., Chen, W., Zeng, X., Yin, W., Gao, X., zhou, c., Sedelain, M., Sun, J.. 2021-01-02. Cas9-cleavage sequences in minimal plasmids enhance non-viral genome targeting of CARs in primary human T cells. https://doi.org/10.1101/2020.12.31.424920

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