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Orthogonal Cas9 proteins for RNA-guided gene regulation and editing.

ABSTRACT: The Cas9 protein from the Streptococcus pyogenes CRISPR-Cas acquired immune system has been adapted for both RNA-guided genome editing and gene regulation in a variety of organisms, but it can mediate only a single activity at a time within any given cell. Here we characterize a set of fully orthogonal Cas9 proteins and demonstrate their ability to mediate simultaneous and independently targeted gene regulation and editing in bacteria and in human cells. We find that Cas9 orthologs display consistent patterns in their recognition of target sequences, and we identify an unexpectedly versatile Cas9 protein from Neisseria meningitidis. We provide a basal set of orthogonal RNA-guided proteins for controlling biological systems and establish a general methodology for characterizing additional proteins.

SUBMITTER: Esvelt KM

PROVIDER: S-EPMC3844869 | biostudies-literature | 2013 Nov

REPOSITORIES: biostudies-literature

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Orthogonal Cas9 proteins for RNA-guided gene regulation and editing.

Esvelt Kevin M KM Mali Prashant P Braff Jonathan L JL Moosburner Mark M Yaung Stephanie J SJ Church George M GM

Nature methods 20130929 11

The Cas9 protein from the Streptococcus pyogenes CRISPR-Cas acquired immune system has been adapted for both RNA-guided genome editing and gene regulation in a variety of organisms, but it can mediate only a single activity at a time within any given cell. Here we characterize a set of fully orthogonal Cas9 proteins and demonstrate their ability to mediate simultaneous and independently targeted gene regulation and editing in bacteria and in human cells. We find that Cas9 orthologs display consi ...[more]

PMID: 24076762

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