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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Vydáno v:	Nature methods Ročník 12; číslo 5; s. 401 - 403
Hlavní autoři:	Kearns, Nicola A, Pham, Hannah, Tabak, Barbara, Genga, Ryan M, Silverstein, Noah J, Garber, Manuel, Maehr, René
Médium:	Journal Article
Jazyk:	angličtina
Vydáno:	New York Nature Publishing Group US 01.05.2015 Nature Publishing Group
Témata:	13 13/100 38/47 45 631/1647/1513/1967 631/1647/2210/2212 631/532/2064/2117 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
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Shrnutí:	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.
Bibliografie:	SourceType-Scholarly Journals-1 ObjectType-Feature-1 content type line 14 ObjectType-Article-1 ObjectType-Feature-2 content type line 23
ISSN:	1548-7091 1548-7105 1548-7105
DOI:	10.1038/nmeth.3325