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Nat Commun. 2019 Feb 13;10(1):729. doi: 10.1038/s41467-019-08736-7.

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

Author information

1
Department of Molecular, Cell, and Developmental Biology, University of California, Los Angeles, CA, 90095, USA.
2
Department of Human Genetics, David Geffen School of Medicine, University of California, Los Angeles, CA, 90095, USA.
3
Molecular Biology Institute, University of California, Los Angeles, CA, 90095, USA.
4
Department of Molecular, Cell, and Developmental Biology, University of California, Los Angeles, CA, 90095, USA. jacobsen@ucla.edu.
5
Howard Hughes Medical Institute, University of California, Los Angeles, CA, 90095, USA. jacobsen@ucla.edu.

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.

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