Genome Editing With CRISPR-Cas12a
The recently-characterized Cas12a (Cpf1) nuclease from type V microbial CRISPR-Cas systems offers new opportunities in genome editing beyond Cas9. However, the utility of Cas12a is limited by its requirement of a TTTV sequence motif in the DNA substrate. Here we use a structure-guided approach to alter the DNA-binding interface of Cas12a, introducing new base-specific interactions with non-canonical DNA sequence motifs. These modifications collectively expand the targeting range of Cas12a by three-fold and increase its utility for genome editing.