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CRISPR-based tools for targeted transcriptional and epigenetic regulation in plants
Umeå University, Faculty of Science and Technology, Umeå Plant Science Centre (UPSC). Umeå University, Faculty of Science and Technology, Department of Plant Physiology.ORCID iD: 0000-0003-0333-3524
Umeå University, Faculty of Science and Technology, Department of Plant Physiology. Umeå University, Faculty of Science and Technology, Umeå Plant Science Centre (UPSC).
Umeå University, Faculty of Science and Technology, Umeå Plant Science Centre (UPSC). Umeå University, Faculty of Science and Technology, Department of Plant Physiology. Max Planck Institute for Developmental Biology, Department of Molecular Biology, Tu ¨bingen, Germany; Beijing Advanced Innovation Centre for Tree Breeding by Molecular Design, Beijing Forestry University, Beijing, People's Republic of China.ORCID iD: 0000-0002-0068-2967
2019 (English)In: PLoS ONE, E-ISSN 1932-6203, Vol. 14, no 9, article id e0222778Article in journal (Refereed) Published
Abstract [en]

Programmable gene regulators that can modulate the activity of selected targets in trans are a useful tool for probing and manipulating gene function. CRISPR technology provides a convenient method for gene targeting that can also be adapted for multiplexing and other modifications to enable strong regulation by a range of different effectors. We generated a vector toolbox for CRISPR/dCas9-based targeted gene regulation in plants, modified with the previously described MS2 system to amplify the strength of regulation, and using Golden Gate-based cloning to enable rapid vector assembly with a high degree of flexibility in the choice of promoters, effectors and targets. We tested the system using the floral regulator FLOWERING LOCUS T (FT) as a target and a range of different effector domains including the transcriptional activator VP64, the H3K27 acetyltransferase p300 and the H3K9 methyltransferase KRYPTONITE. When transformed into Arabidopsis thaliana, several of the constructs caused altered flowering time phenotypes that were associated with changes in FT expression and/or epigenetic status, thus demonstrating the effectiveness of the system. The MS2-CRISPR/dCas9 system can be used to modulate transcriptional activity and epigenetic status of specific target genes in plants, and provides a versatile tool that can easily be used with different targets and types of regulation for a range of applications.

Place, publisher, year, edition, pages
Public Library of Science , 2019. Vol. 14, no 9, article id e0222778
National Category
Botany
Identifiers
URN: urn:nbn:se:umu:diva-166461DOI: 10.1371/journal.pone.0222778ISI: 000489744000001PubMedID: 31557222Scopus ID: 2-s2.0-85072661389OAI: oai:DiVA.org:umu-166461DiVA, id: diva2:1379454
Funder
Knut and Alice Wallenberg Foundation, KAW 2016.0025Available from: 2019-12-17 Created: 2019-12-17 Last updated: 2019-12-17Bibliographically approved

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