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

• Published in: Nature communications Vol. 10; no. 1; pp. 729 - 11
• Main Authors: Papikian, Ashot, Liu, Wanlu, Gallego-Bartolomé, Javier, Jacobsen, Steven E.
• Format: Journal Article
• Language: English
• Published: London Nature Publishing Group UK 13.02.2019; Nature Publishing Group; Nature Portfolio
• Subjects: 38/1; 38/15; 38/22; 38/23; 38/35; 38/39; 38/44; 38/77; 38/88; 38/90; 38/91; 45/15; 45/22; 45/23; 45/44; 45/77; 45/88; 45/90; 45/91; 631/337/4041/3196; 631/449/2491/2046; 631/61/447/2311; Activation; Arabidopsis; Arabidopsis - genetics; Arabidopsis - metabolism; Arabidopsis Proteins - genetics; Arabidopsis Proteins - metabolism; Binding Sites - genetics; Catalysis; Chromatin; Chromatin - genetics; Chromatin - metabolism; CRISPR; CRISPR-Cas Systems; Deoxyribonucleic acid; DNA; DNA Methylation; DNA methyltransferase; DRM protein; Epigenesis, Genetic; Gene Editing - methods; Gene Expression Regulation, Plant; Homeodomain Proteins - genetics; Homeodomain Proteins - metabolism; Humanities and Social Sciences; Loci; multidisciplinary; Phenotypes; Plants, Genetically Modified; Promoter Regions, Genetic - genetics; Proteins; Science; Science (multidisciplinary); Transcription; Transcription Factors - genetics; Transcription Factors - metabolism
• ISSN: 2041-1723; 2041-1723
• DOI: 10.1038/s41467-019-08736-7

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. Few approaches for targeted manipulation of the epigenome are available in plants. Here, the authors adapt the dCas9-SunTag system to engineer targeted gene activation and site-specific manipulation of DNA methylation in Arabidopsis .