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Engineering a high-fidelity SpaCas9 variant
(A) Schematic of designed SpaCas9-HF. R266A, N281D, and Q445A are all located in the REC2 domain. (B) Comparison of the tolerance of SpaCas9 and SpaCas9-HF for mismatched sgRNAs. Mismatched sgRNAs that differed from the S2 site by two or three nucleotides. Mismatched nucleotides and the PAM sequence are shown in red and green, respectively. Error bars indicate the SEM (n = 3).
