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. 2020 Oct;38(10):1154-1158.
doi: 10.1038/s41587-020-0517-0. Epub 2020 May 11.

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

Pranam Chatterjee #  1   2 , Noah Jakimo #  3   4 , Jooyoung Lee  5 , Nadia Amrani  5 , Tomás Rodríguez  5 , Sabrina R T Koseki  3   4 , Emma Tysinger  3   4 , Rui Qing  4 , Shilei Hao  4 , Erik J Sontheimer  5 , Joseph Jacobson  3   4
Affiliations

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

Pranam Chatterjee et al. Nat Biotechnol. 2020 Oct.

Erratum in

Abstract

CRISPR enzymes require a protospacer-adjacent motif (PAM) near the target cleavage site, constraining the sequences accessible for editing. In the present study, we combine protein motifs from several orthologs to engineer two variants of Streptococcus canis Cas9-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 use of CRISPR editing for diverse applications.

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Comment in

  • PAM-less is more.
    Tang L. Tang L. Nat Methods. 2020 Jun;17(6):559. doi: 10.1038/s41592-020-0861-5. Nat Methods. 2020. PMID: 32499620 No abstract available.

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