1. Highly efficient multiplex human T cell engineering without double-strand breaks using Cas9 base editors.
- Author
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Webber, Beau R., Lonetree, Cara-lin, Kluesner, Mitchell G., Johnson, Matthew J., Pomeroy, Emily J., Diers, Miechaleen D., Lahr, Walker S., Draper, Garrett M., Slipek, Nicholas J., Smeester, Branden A., Lovendahl, Klaus N., McElroy, Amber N., Gordon, Wendy R., Osborn, Mark J., and Moriarity, Branden S.
- Subjects
GENETIC disorder treatment ,CHIMERIC antigen receptors ,HUMAN T cells ,CELLULAR therapy ,NUCLEASE genetics - Abstract
The fusion of genome engineering and adoptive cellular therapy holds immense promise for the treatment of genetic disease and cancer. Multiplex genome engineering using targeted nucleases can be used to increase the efficacy and broaden the application of such therapies but carries safety risks associated with unintended genomic alterations and genotoxicity. Here, we apply base editor technology for multiplex gene modification in primary human T cells in support of an allogeneic CAR-T platform and demonstrate that base editor can mediate highly efficient multiplex gene disruption with minimal double-strand break induction. Importantly, multiplex base edited T cells exhibit improved expansion and lack double strand break-induced translocations observed in T cells edited with Cas9 nuclease. Our findings highlight base editor as a powerful platform for genetic modification of therapeutically relevant primary cell types. Multiplexed genome engineering with Cas9 can increase efficiency but also the risk of unintended alterations. Here the authors demonstrate the use of multiplexed base editors on primary T cells with reduced translocation frequency. [ABSTRACT FROM AUTHOR]
- Published
- 2019
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