Orthogonal gene knockout and activation with a catalytically active Cas9 nuclease
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Zhang_Orthogonal gene.pdf
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Author(s)
Joung, Julia
Zhang, Feng
Konermann, Silvana
Dahlman, James E.
Abudayyeh, Omar Osama
Gootenberg, Jonathan S.
Date Issued
October 2015
Journal
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.
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Author's final manuscript
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.
MIT Department
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 MIT
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DOI of Published Version
https://doi.org/10.1038/nbt.3390
