Cas12 proteins and uses thereof

Abstract

The present disclosure relates to Cas12 proteins and uses thereof. In certain embodiments, a Cas12 protein is provided whose amino acid sequence has at least 50% sequence identity to SEQ ID NO: 1 and includes one, two, or more amino acid differences relative to SEQ ID NO: 1. In certain embodiments, the Cas12 protein has at least 50% sequence identity to SEQ ID NO: 18. In certain embodiments, the Cas12 protein recognizes a PAM sequence of A. In certain embodiments, gene-editing efficiency of a wild-type Cas12 protein in mammalian cells is improved by introducing rational and irrational amino acid mutations. In certain embodiments, the Cas12 protein exhibits increased editing efficiency and/or a reduced off-target rate and, by virtue of recognizing a single-base A PAM, enables editing of target sequences that were previously difficult to edit, thereby substantially expanding the range of editable targets.

Dependencies are inferred from the claim text.