Orthogonal gene knockout and activation with a catalytically active Cas9 nuclease
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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.
Date issued
2015-10Department
Harvard University--MIT Division of Health Sciences and Technology; Massachusetts Institute of Technology. Department of Biological Engineering; Massachusetts Institute of Technology. Department of Brain and Cognitive Sciences; McGovern Institute for Brain Research at MITJournal
Nature Biotechnology
Publisher
Nature Publishing Group
Citation
Dahlman, James E, Omar O Abudayyeh, Julia Joung, Jonathan S Gootenberg, Feng Zhang, and Silvana Konermann. “Orthogonal Gene Knockout and Activation with a Catalytically Active Cas9 Nuclease.” Nature Biotechnology 33, no. 11 (October 5, 2015): 1159–61.
Version: Author's final manuscript
ISSN
1087-0156
1546-1696