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

Kearns, Nicola A; Pham, Hannah; Tabak, Barbara; Genga, Ryan M; Silverstein, Noah J; Garber, Manuel; Maehr, René

Kearns, Nicola A; Pham, Hannah; Tabak, Barbara; Genga, Ryan M; Silverstein, Noah J; Garber, Manuel; Maehr, René.

Afiliación

Kearns NA; Diabetes Center of Excellence, University of Massachusetts Medical School, Worcester, Massachusetts 01605, USA.
Pham H; Program in Molecular Medicine, University of Massachusetts Medical School, Worcester, Massachusetts 01605, USA.
Tabak B; Diabetes Center of Excellence, University of Massachusetts Medical School, Worcester, Massachusetts 01605, USA.
Genga RM; Program in Molecular Medicine, University of Massachusetts Medical School, Worcester, Massachusetts 01605, USA.
Silverstein NJ; Program in Molecular Medicine, University of Massachusetts Medical School, Worcester, Massachusetts 01605, USA.
Garber M; Bioinformatics and Integrative Biology, University of Massachusetts Medical School, Worcester, MA 01605, USA.
Maehr R; Diabetes Center of Excellence, University of Massachusetts Medical School, Worcester, Massachusetts 01605, USA.

Nat Methods ; 12(5): 401-403, 2015 May.

Article en En | MEDLINE | ID: mdl-25775043

ABSTRACT

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.

Asunto(s)

Caspasa 9/metabolismo; Elementos de Facilitación Genéticos/fisiología; Histona Demetilasas/metabolismo; Animales; Proteínas Bacterianas/genética; Proteínas Bacterianas/metabolismo; Caspasa 9/efectos de los fármacos; Células Cultivadas; Células Madre Embrionarias/metabolismo; Elementos de Facilitación Genéticos/genética; Regulación Enzimológica de la Expresión Génica; Genes Reporteros; Histona Demetilasas/efectos de los fármacos; Histonas/metabolismo; Ratones; Neisseria meningitidis/enzimología; Proteínas Recombinantes/metabolismo

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Texto completo: 1 Banco de datos: MEDLINE Asunto principal: Elementos de Facilitación Genéticos / Caspasa 9 / Histona Demetilasas Límite: Animals Idioma: En Revista: Nat Methods Asunto de la revista: TECNICAS E PROCEDIMENTOS DE LABORATORIO Año: 2015 Tipo del documento: Article País de afiliación: Estados Unidos

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