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Functional annotation of native enhancers with a Cas9-histone demethylase fusion.

ABSTRACT: Understanding of mammalian enhancers is limited by the lack of a technology to rapidly and thoroughly test the cell type-specific function. Here, we use a nuclease-deficient Cas9 (dCas9)-histone demethylase fusion to functionally characterize previously described and new enhancer elements for their roles in the embryonic stem cell state. Further, we distinguish the mechanism of action of dCas9-LSD1 at enhancers from previous dCas9-effectors.

SUBMITTER: Kearns NA 

PROVIDER: S-EPMC4414811 | biostudies-literature | 2015 May

REPOSITORIES: biostudies-literature

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Functional annotation of native enhancers with a Cas9-histone demethylase fusion.

Kearns Nicola A NA   Pham Hannah H   Tabak Barbara B   Genga Ryan M RM   Silverstein Noah J NJ   Garber Manuel M   Maehr René R  

Nature methods 20150316 5

Understanding of mammalian enhancers is limited by the lack of a technology to rapidly and thoroughly test the cell type-specific function. Here, we use a nuclease-deficient Cas9 (dCas9)-histone demethylase fusion to functionally characterize previously described and new enhancer elements for their roles in the embryonic stem cell state. Further, we distinguish the mechanism of action of dCas9-LSD1 at enhancers from previous dCas9-effectors. ...[more]

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