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

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.

Citation

James E Dahlman, Omar O Abudayyeh, Julia Joung, Jonathan S Gootenberg, Feng Zhang, Silvana Konermann. Orthogonal gene knockout and activation with a catalytically active Cas9 nuclease. Nature biotechnology. 2015 Nov;33(11):1159-61

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PMID: 26436575

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