Highly precise genome editing using enhanced CRISPR-Cas12a nickase module

Abstract

AbstractThe CRISPR-Cas nickase system for genome editing has attracted considerable attention owing to its safety, efficiency, and versatility. Although alternative effectors to Cas9 have the potential to expand the scope of genome editing, their application has not been optimized. Herein, we used an enhanced CRISPR-Cas12a nickase system to induce mutations by targeting genes in a human-derived cell line. The newly developed CRISPR-Cas12a nickase system effectively introduced mutations into target genes under a specific directionality and distance between nickases. In particular, the dual-mode Cas12a nickase system can improve the target specificity by 10.5- to 12.5-fold over that of double-strand breaks induced by wild-type Cas12a. By effectively inducing mutations in the target gene in single- or dual-mode, Cas12a nickase addresses the limitations of Cas9 nickase and is expected to contribute to the development of future genome editing technologies.