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

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 DN...

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Autores principales: Papikian, Ashot, Liu, Wanlu, Gallego-Bartolomé, Javier, Jacobsen, Steven E.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Nature Publishing Group UK 2019
Materias:
Article
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6374409/
https://www.ncbi.nlm.nih.gov/pubmed/30760722
http://dx.doi.org/10.1038/s41467-019-08736-7
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author Papikian, Ashot
Liu, Wanlu
Gallego-Bartolomé, Javier
Jacobsen, Steven E.
author_facet Papikian, Ashot
Liu, Wanlu
Gallego-Bartolomé, Javier
Jacobsen, Steven E.
author_sort Papikian, Ashot
collection PubMed
description 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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spelling pubmed-63744092019-02-15 Site-specific manipulation of Arabidopsis loci using CRISPR-Cas9 SunTag systems Papikian, Ashot Liu, Wanlu Gallego-Bartolomé, Javier Jacobsen, Steven E. Nat Commun Article 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. Nature Publishing Group UK 2019-02-13 /pmc/articles/PMC6374409/ /pubmed/30760722 http://dx.doi.org/10.1038/s41467-019-08736-7 Text en © The Author(s) 2019 Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.
spellingShingle Article
Papikian, Ashot
Liu, Wanlu
Gallego-Bartolomé, Javier
Jacobsen, Steven E.
Site-specific manipulation of Arabidopsis loci using CRISPR-Cas9 SunTag systems
title Site-specific manipulation of Arabidopsis loci using CRISPR-Cas9 SunTag systems
title_full Site-specific manipulation of Arabidopsis loci using CRISPR-Cas9 SunTag systems
title_fullStr Site-specific manipulation of Arabidopsis loci using CRISPR-Cas9 SunTag systems
title_full_unstemmed Site-specific manipulation of Arabidopsis loci using CRISPR-Cas9 SunTag systems
title_short Site-specific manipulation of Arabidopsis loci using CRISPR-Cas9 SunTag systems
title_sort site-specific manipulation of arabidopsis loci using crispr-cas9 suntag systems
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6374409/
https://www.ncbi.nlm.nih.gov/pubmed/30760722
http://dx.doi.org/10.1038/s41467-019-08736-7
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