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Nat Methods. 2015 May;12(5):401-403. doi: 10.1038/nmeth.3325. Epub 2015 Mar 16.

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

Author information

1
Diabetes Center of Excellence, University of Massachusetts Medical School, Worcester, Massachusetts 01605, USA.
2
Program in Molecular Medicine, University of Massachusetts Medical School, Worcester, Massachusetts 01605, USA.
3
Bioinformatics and Integrative Biology, University of Massachusetts Medical School, Worcester, MA 01605, USA.
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Contributed equally

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.

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PMID:
25775043
PMCID:
PMC4414811
DOI:
10.1038/nmeth.3325
[Indexed for MEDLINE]
Free PMC Article

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