Inducible and multiplex gene regulation using CRISPR-Cpf1-based transcription factors.

Tak, Y Esther; Kleinstiver, Benjamin P; Nuñez, James K; Hsu, Jonathan Y; Horng, Joy E; Gong, Jingyi; Weissman, Jonathan S; Joung, J Keith

Tak, Y Esther; Kleinstiver, Benjamin P; Nuñez, James K; Hsu, Jonathan Y; Horng, Joy E; Gong, Jingyi; Weissman, Jonathan S; Joung, J Keith.

Afiliación

Tak YE; Molecular Pathology Unit, Center for Cancer Research, and Center for Computational and Integrative Biology, Massachusetts General Hospital, Charlestown, Massachusetts, USA.
Kleinstiver BP; Department of Pathology, Harvard Medical School, Boston, Massachusetts, USA.
Nuñez JK; Molecular Pathology Unit, Center for Cancer Research, and Center for Computational and Integrative Biology, Massachusetts General Hospital, Charlestown, Massachusetts, USA.
Hsu JY; Department of Pathology, Harvard Medical School, Boston, Massachusetts, USA.
Horng JE; Department of Cellular & Molecular Pharmacology University of California, San Francisco, San Francisco, California, USA.
Gong J; Molecular Pathology Unit, Center for Cancer Research, and Center for Computational and Integrative Biology, Massachusetts General Hospital, Charlestown, Massachusetts, USA.
Weissman JS; Department of Biological Engineering, Massachusetts Institute of Technology, Cambridge, Massachusetts, USA.
Joung JK; Molecular Pathology Unit, Center for Cancer Research, and Center for Computational and Integrative Biology, Massachusetts General Hospital, Charlestown, Massachusetts, USA.

Nat Methods ; 14(12): 1163-1166, 2017 Dec.

Article en En | MEDLINE | ID: mdl-29083402

ABSTRACT

ABSTRACT

Targeted and inducible regulation of mammalian gene expression is a broadly important capability. We engineered drug-inducible catalytically inactive Cpf1 nuclease fused to transcriptional activation domains to tune the expression of endogenous genes in human cells. Leveraging the multiplex capability of the Cpf1 platform, we demonstrate both synergistic and combinatorial gene expression in human cells. Our work should enable the development of multiplex gene perturbation library screens for understanding complex cellular phenotypes.

Asunto(s)

Proteínas Bacterianas/genética; Sistemas CRISPR-Cas/genética; Repeticiones Palindrómicas Cortas Agrupadas y Regularmente Espaciadas/genética; Endonucleasas/genética; Activación Transcripcional; Técnicas de Cultivo de Célula; Proteínas Fluorescentes Verdes/genética; Células HEK293; Proteína Vmw65 de Virus del Herpes Simple/genética; Humanos; Proteínas Inmediatas-Precoces/genética; Plásmidos; Proteínas Recombinantes de Fusión/genética; Transactivadores/genética; Factor de Transcripción ReIA/genética; Transfección

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Texto completo: 1 Colección: 01-internacional Banco de datos: MEDLINE Asunto principal: Proteínas Bacterianas / Activación Transcripcional / Endonucleasas / Repeticiones Palindrómicas Cortas Agrupadas y Regularmente Espaciadas / Sistemas CRISPR-Cas Límite: Humans Idioma: En Revista: Nat Methods Asunto de la revista: TECNICAS E PROCEDIMENTOS DE LABORATORIO Año: 2017 Tipo del documento: Article País de afiliación: Estados Unidos

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