CRISPR-Cas12a REC2 - NUC interactions drive target-strand cleavage and constrain trans cleavage.

ABSTRACT: CRISPR-Cas12a effects RNA-guided cleavage of dsDNA in cis, after which it remains catalytically active and non-specifically cleaves ssDNA in trans. Native host-defence by Cas12a employs cis cleavage, which can be repurposed for the genome editing of other organisms, and trans cleavage can be used for in vitro DNA detection. Cas12a orthologues have high structural similarity and a conserved mechanism of DNA cleavage, yet highly different efficacies when applied for genome editing or DNA detection. By comparing three well characterised Cas12a orthologues (FnCas12a, LbCas12a, and AsCas12a), we sought to determine what drives their different cis and trans cleavage, and how this relates to their applied function. We integrated in vitro DNA

SUBMITTER: Newman A 

PROVIDER: S-EPMC11974684 | biostudies-literature | 2025 Mar

REPOSITORIES: biostudies-literature