CRISPR-Cas12a exhibits metal-dependent specificity switching.

ABSTRACT: Cas12a is the immune effector of type V-A CRISPR-Cas systems and has been co-opted for genome editing and other biotechnology tools. The specificity of Cas12a has been the subject of extensive investigation both in vitro and in genome editing experiments. However, in vitro studies have often been performed at high magnesium ion concentrations that are inconsistent with the free Mg²⁺ concentrations that would be present in cells. By profiling the specificity of Cas12a orthologs at a range of Mg²⁺ concentrations, we find that Cas12a switches its specificity depending on metal ion concentration. Lowering Mg²⁺ concentration decreases cleavage defects caused by seed mismatches, while increasing the defects caused by PAM-distal mismatches. We show t

SUBMITTER: Nguyen GT 

PROVIDER: S-EPMC10705449 | biostudies-literature | 2024 Jan

REPOSITORIES: biostudies-literature