Site-specific manipulation of Arabidopsis loci using CRISPR-Cas9 SunTag systems.

Papikian, Ashot; Liu, Wanlu; Gallego-Bartolomé, Javier; Jacobsen, Steven E

Papikian, Ashot; Liu, Wanlu; Gallego-Bartolomé, Javier; Jacobsen, Steven E.

Afiliación

Papikian A; Department of Molecular, Cell, and Developmental Biology, University of California, Los Angeles, CA, 90095, USA.
Liu W; Department of Human Genetics, David Geffen School of Medicine, University of California, Los Angeles, CA, 90095, USA.
Gallego-Bartolomé J; Department of Molecular, Cell, and Developmental Biology, University of California, Los Angeles, CA, 90095, USA.
Jacobsen SE; Molecular Biology Institute, University of California, Los Angeles, CA, 90095, USA.

Nat Commun ; 10(1): 729, 2019 02 13.

Article en En | MEDLINE | ID: mdl-30760722

ABSTRACT

ABSTRACT

Understanding genomic functions requires site-specific manipulation of loci via efficient protein effector targeting systems. However, few approaches for targeted manipulation of the epigenome are available in plants. Here, we adapt the dCas9-SunTag system to engineer targeted gene activation and DNA methylation in Arabidopsis. We demonstrate that a dCas9-SunTag system utilizing the transcriptional activator VP64 drives robust and specific activation of several loci, including protein coding genes and transposable elements, in diverse chromatin contexts. In addition, we present a CRISPR-based methylation targeting system for plants, utilizing a SunTag system with the catalytic domain of the Nicotiana tabacum DRM methyltransferase, which efficiently targets DNA methylation to specific loci, including the FWA promoter, triggering a developmental phenotype, and the SUPERMAN promoter. These SunTag systems represent valuable tools for the site-specific manipulation of plant epigenomes.

Asunto(s)

Proteínas de Arabidopsis/genética; Arabidopsis/genética; Sistemas CRISPR-Cas; Metilación de ADN; Edición Génica/métodos; Arabidopsis/metabolismo; Proteínas de Arabidopsis/metabolismo; Sitios de Unión/genética; Cromatina/genética; Cromatina/metabolismo; Epigénesis Genética; Regulación de la Expresión Génica de las Plantas; Proteínas de Homeodominio/genética; Proteínas de Homeodominio/metabolismo; Plantas Modificadas Genéticamente; Regiones Promotoras Genéticas/genética; Factores de Transcripción/genética; Factores de Transcripción/metabolismo

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Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Arabidopsis / Metilación de ADN / Proteínas de Arabidopsis / Sistemas CRISPR-Cas / Edición Génica Idioma: En Revista: Nat Commun Asunto de la revista: BIOLOGIA / CIENCIA Año: 2019 Tipo del documento: Article País de afiliación: Estados Unidos

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