Functional annotation of native enhancers with a Cas9–histone demethylase fusion

Targeting a histone demethylase to specific enhancers with Cas9 allows modification of epigenetic marks at the enhancers. 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...

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Bibliographic Details
Published in:Nature methods Vol. 12; no. 5; pp. 401 - 403
Main Authors: Kearns, Nicola A, Pham, Hannah, Tabak, Barbara, Genga, Ryan M, Silverstein, Noah J, Garber, Manuel, Maehr, René
Format: Journal Article
Language:English
Published: New York Nature Publishing Group US 01.05.2015
Nature Publishing Group
Subjects:
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Animals
Bacterial Proteins - genetics
Bacterial Proteins - metabolism
Bioinformatics
Biological Microscopy
Biological Techniques
Biomedical Engineering/Biotechnology
brief-communication
Caspase 9 - drug effects
Caspase 9 - metabolism
Cells, Cultured
Cellular biology
Cellular control mechanisms
Embryonic Stem Cells - metabolism
Enhancer Elements, Genetic - genetics
Enhancer Elements, Genetic - physiology
Enzymes
Gene Expression Regulation, Enzymologic
Genes, Reporter
Genomics
Histone Demethylases - drug effects
Histone Demethylases - metabolism
Histones
Histones - metabolism
Life Sciences
Mammals
Mice
Neisseria meningitidis - enzymology
Physiological aspects
Proteomics
Recombinant Proteins - metabolism
Stem cells
ISSN:1548-7091, 1548-7105, 1548-7105
Online Access:Get full text
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Summary:Targeting a histone demethylase to specific enhancers with Cas9 allows modification of epigenetic marks at the enhancers. 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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ISSN:1548-7091
1548-7105
1548-7105
DOI:10.1038/nmeth.3325