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Orthogonal gene knockout and activation with a catalytically active Cas9 nuclease.

ABSTRACT: We have developed a CRISPR-based method that uses catalytically active Cas9 and distinct single guide (sgRNA) constructs to knock out and activate different genes in the same cell. These sgRNAs, with 14- to 15-bp target sequences and MS2 binding loops, can activate gene expression using an active Streptococcus pyogenes Cas9 nuclease, without inducing double-stranded breaks. We use these 'dead RNAs' to perform orthogonal gene knockout and transcriptional activation in human cells.

SUBMITTER: Dahlman JE

PROVIDER: S-EPMC4747789 | biostudies-literature | 2015 Nov

REPOSITORIES: biostudies-literature

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Orthogonal gene knockout and activation with a catalytically active Cas9 nuclease.

Dahlman James E JE Abudayyeh Omar O OO Joung Julia J Gootenberg Jonathan S JS Zhang Feng F Konermann Silvana S

Nature biotechnology 20151101 11

We have developed a CRISPR-based method that uses catalytically active Cas9 and distinct single guide (sgRNA) constructs to knock out and activate different genes in the same cell. These sgRNAs, with 14- to 15-bp target sequences and MS2 binding loops, can activate gene expression using an active Streptococcus pyogenes Cas9 nuclease, without inducing double-stranded breaks. We use these 'dead RNAs' to perform orthogonal gene knockout and transcriptional activation in human cells. ...[more]

PMID: 26436575

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