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Orthogonal gene knock out and activation with a catalytically active Cas9 nuclease
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Published Date:Nov 2015
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Source:Nat Biotechnol. 33(11):1159-1161.
[PDF - 398.15 KB]
Details:
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Personal Authors:
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Keywords:
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Pubmed ID:26436575
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Pubmed Central ID:PMC4747789
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Funding:1DP1-MH100706/DP/NCCDPHP CDC HHS/United States
DP1 MH100706/MH/NIMH NIH HHS/United States
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Document Type:
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Description: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.
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Supporting Files:
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