Engineered CRISPR-Cas9 nuclease with expanded targeting space

Abstract

Expanding the targeting space of Cas9 CRISPR-Cas9 associates with a guide RNA to target and cleave a specific DNA site next to a protospacer adjacent motif (PAM). Streptococcus pyogenes Cas9 (SpCas9), the one most often used for genome editing, only recognizes the NGG sequence (where N is any nucleobase) as the PAM, which restricts regions in the genome that can be targeted. To address this limitation, Nishimasu et al. created a SpCas9 variant that recognizes NG rather than NGG. The SpCas9-NG variant increased the targeting range, had a specificity similar to that of the wild-type enzyme, and could be used with a base editor. Thus, SpCas9-NG is a powerful addition to the CRISPR-Cas9 genome engineering toolbox and will be useful in a broad range of applications, from basic research to clinical therapeutics. Science , this issue p. 1259

Keywords

Affiliated Institutions

• Keio University Shonan Fujisawa JP
• The University of Tokyo JP
• Broad Institute US
• The University of Osaka JP
• Massachusetts Institute of Technology US
• McGovern Institute for Brain Research US
• Keio University JP

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