An engineered ScCas9 with broad PAM range and high specificity and activity

Pranam Chatterjee


 CRISPR enzymes require a protospacer adjacent motif (PAM) near the target cleavage site, constraining the sequences accessible for editing. Here, we combine protein motifs from several orthologs to engineer two variants of Streptococcus canis Cas9 (ScCas9) – Sc++ and a higher-fidelity mutant HiFi-Sc++ – that have simultaneously broad 5'-NNG-3' PAM compatibility, robust DNA cleavage activity, and minimal off-target activity. Sc++ and HiFi-Sc++ extend the utility of CRISPR editing for diverse applications.

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