Orthogonal Cas9 proteins for RNA-guided gene regulation and editing

Esvelt et al. (2013). Nat Methods DOI: 10.1038/nmeth.2681 Citations: 814

SpCas9 could mediate only one RNA-guided activity per cell. This paper found orthogonal Cas9 proteins with distinct PAMs, including a targetable ortholog from N. meningitidis. NmCas9 repressed and activated genes in human cells without cross-talk with SpCas9.

Parts used

NmeCas9 Nuclease

Key findings

  • Characterized multiple Cas9 orthologs including NmeCas9 from Neisseria meningitidis. Demonstrated orthogonal functionality for multiplex applications. Established NmeCas9 compact size for AAV delivery.