Engineered CRISPR-Cas12a variants with increased activities and improved targeting ranges for gene, epigenetic and base editing.

ABSTRACT: Broad use of CRISPR-Cas12a (formerly Cpf1) nucleases¹ has been hindered by the requirement for an extended TTTV protospacer adjacent motif (PAM)². To address this limitation, we engineered an enhanced Acidaminococcus sp. Cas12a variant (enAsCas12a) that has a substantially expanded targeting range, enabling targeting of many previously inaccessible PAMs. On average, enAsCas12a exhibits a twofold higher genome editing activity on sites with canonical TTTV PAMs compared to wild-type AsCas12a, and we successfully grafted a subset of mutations from enAsCas12a onto other previously described AsCas12a variants³ to enhance their activities. enAsCas12a improves the efficiency of multiplex gene editing, endogenous gene activation and C-to-T base editing, and we engi

SUBMITTER: Kleinstiver BP 

PROVIDER: S-EPMC6401248 | biostudies-literature | 2019 Mar

REPOSITORIES: biostudies-literature