Site-specific manipulation of Arabidopsis loci using CRISPR-Cas9 SunTag systems.
basic_science · Level V
Where this comes from
- Record sourced from PubMed, PMID 30760722.
- Also identified by DOI 10.1038/s41467-019-08736-7 and PMC identifier 6374409.
- Licence recorded as CC BY.
- The licence permits redistribution, so the abstract is shown in full and the full text is available from the publisher.
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.
Medical subject headings
- Arabidopsis
- Arabidopsis Proteins
- CRISPR-Cas Systems
- DNA Methylation
- Gene Editing