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Zhang, Liyang; Zuris, John A; Viswanathan, Ramya; Edelstein, Jasmine N; Turk, Rolf; Thommandru, Bernice; Rube, H Tomas; Glenn, Steve E; Collingwood, Michael A; Bode, Nicole M; Beaudoin, Sarah F; Lele, Swarali; Scott, Sean N; Wasko, Kevin M; Sexton, Steven; Borges, Christopher M; Schubert, Mollie S; Kurgan, Gavin L; McNeill, Matthew S; Fernandez, Cecilia A; Myer, Vic E; Morgan, Richard A; Behlke, Mark A; Vakulskas, Christopher A
Nature communications, 06/2021, Letnik: 12, Številka: 1Journal Article
Though AsCas12a fills a crucial gap in the current genome editing toolbox, it exhibits relatively poor editing efficiency, restricting its overall utility. Here we isolate an engineered variant, "AsCas12a Ultra", that increased editing efficiency to nearly 100% at all sites examined in HSPCs, iPSCs, T cells, and NK cells. We show that AsCas12a Ultra maintains high on-target specificity thereby mitigating the risk for off-target editing and making it ideal for complex therapeutic genome editing applications. We achieved simultaneous targeting of three clinically relevant genes in T cells at >90% efficiency and demonstrated transgene knock-in efficiencies of up to 60%. We demonstrate site-specific knock-in of a CAR in NK cells, which afforded enhanced anti-tumor NK cell recognition, potentially enabling the next generation of allogeneic cell-based therapies in oncology. AsCas12a Ultra is an advanced CRISPR nuclease with significant advantages in basic research and in the production of gene edited cell medicines.
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JCR | SNIP | JCR | SNIP | JCR | SNIP | JCR | SNIP |
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Povezave do osebnih bibliografij avtorjev | Povezave do podatkov o raziskovalcih v sistemu SICRIS |
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Vir: Osebne bibliografije
in: SICRIS
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