Functional Annotation of Native Enhancers With a Cas9-histone Demethylase Fusion

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

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.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Bacterial Proteins / genetics
  • Bacterial Proteins / metabolism
  • Caspase 9 / drug effects
  • Caspase 9 / metabolism*
  • Cells, Cultured
  • Embryonic Stem Cells / metabolism
  • Enhancer Elements, Genetic / genetics
  • Enhancer Elements, Genetic / physiology*
  • Gene Expression Regulation, Enzymologic
  • Genes, Reporter
  • Histone Demethylases / drug effects
  • Histone Demethylases / metabolism*
  • Histones / metabolism
  • Mice
  • Neisseria meningitidis / enzymology
  • Recombinant Proteins / metabolism

Substances

  • Bacterial Proteins
  • Histones
  • Recombinant Proteins
  • Histone Demethylases
  • Caspase 9