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- W4220795729 abstract "Numerous rationally-designed and directed-evolution variants of SpCas9 have been reported to expand the utility of CRISPR technology. Here, we assess the activity and specificity of WT-Cas9 and 10 SpCas9 variants by benchmarking their PAM preferences, on-target activity, and off-target susceptibility in cell culture assays with thousands of guides targeting endogenous genes. To enhance the coverage and thus utility of base editing screens, we demonstrate that the SpCas9-NG and SpG variants are compatible with both A > G and C > T base editors, more than tripling the number of guides and assayable residues. We demonstrate the performance of these technologies by screening for loss-of-function mutations in BRCA1 and Venetoclax-resistant mutations in BCL2, identifying both known and new mutations that alter function. We anticipate that the tools and methodologies described here will facilitate the investigation of genetic variants at a finer and deeper resolution for any locus of interest." @default.
- W4220795729 created "2022-04-03" @default.
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- W4220795729 date "2022-03-14" @default.
- W4220795729 modified "2023-10-14" @default.
- W4220795729 title "Benchmarking of SpCas9 variants enables deeper base editor screens of BRCA1 and BCL2" @default.
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- W4220795729 doi "https://doi.org/10.1038/s41467-022-28884-7" @default.
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