Functional annotation of native enhancers with a Cas9-histone demethylase fusion
Diabetes Center of Excellence; Program in Molecular Medicine; Program in Bioinformatics and Integrative Biology
Animals; Bacterial Proteins; Caspase 9; Cells, Cultured; Embryonic Stem Cells; Enhancer Elements, Genetic; Gene Expression Regulation, Enzymologic; Genes, Reporter; Histone Demethylases; Histones; Mice; Neisseria meningitidis; Recombinant Proteins
Biochemistry, Biophysics, and Structural Biology | Bioinformatics | Computational Biology | Integrative Biology | Systems Biology
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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Citation: Nat Methods. 2015 May;12(5):401-3. doi: 10.1038/nmeth.3325. Epub 2015 Mar 16. Link to article on publisher's site
Kearns, Nicola A.; Pham, Hannah; Tabak, Barbara; Genga, Ryan M. J.; Silverstein, Noah J.; Garber, Manuel; and Maehr, Rene, "Functional annotation of native enhancers with a Cas9-histone demethylase fusion" (2015). Program in Bioinformatics and Integrative Biology Publications and Presentations. 79.