CRISPR-based tools for targeted transcriptional and epigenetic regulation in plants


Autoři: Joanne E. Lee aff001;  Manuela Neumann aff002;  Daniel Iglesias Duro aff001;  Markus Schmid aff001
Působiště autorů: Umeå Plant Science Centre, Department of Plant Physiology, Umeå University, Umeå, Sweden aff001;  Max Planck Institute for Developmental Biology, Department of Molecular Biology, Tübingen, Germany aff002;  Beijing Advanced Innovation Centre for Tree Breeding by Molecular Design, Beijing Forestry University, Beijing, People’s Republic of China aff003
Vyšlo v časopise: PLoS ONE 14(9)
Kategorie: Research Article
doi: 10.1371/journal.pone.0222778

Souhrn

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.

Klíčová slova:

Arabidopsis thaliana – Cloning – Epigenetics – Flowering plants – Gene regulation – Genetically modified plants – Histones – Regulator genes


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