Rational designs of Cas12a nuclease with the improved targeting specificity at both the proximal and distal region from the PAM
Our findings provide a novel version of AsCas12a variant with low off-target effects, especially capable of dealing with the high off-targeting at the distal region from the PAM. Overall design: we engineered a hyper fidelity AsCas12a variant carrying the mutations S186A/R301A/T315A/Q1014A/K414A (termed HyperFi-As) by modifying amino acid residues interacting with the target DNA and crRNA strand