Rational designs of Cas12a nuclease with the improved targeting specificity at both the proximal and distal region from the PAM. 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