Publication: Orthogonal gene knock out and activation with a catalytically active Cas9 nuclease
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2015
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Dahlman, James E., Omar O. Abudayyeh, Julia Joung, Jonathan S. Gootenberg, Feng Zhang, and Silvana Konermann. 2015. “Orthogonal gene knock out and activation with a catalytically active Cas9 nuclease.” Nature biotechnology 33 (11): 1159-1161. doi:10.1038/nbt.3390. http://dx.doi.org/10.1038/nbt.3390.
Abstract
We have developed a CRISPR-based method that uses catalytically active Cas9 and distinct sgRNA constructs to knock out and activate different genes in the same cell. These sgRNAs, with 14 15 bp target sequences and MS2 binding loops, can activate gene expression using an active Cas9 nuclease, without inducing DSBs. We use these ‘dead RNAs’ to perform orthogonal gene knockout and transcriptional activation in human cells.
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