CRISPR-based tools for targeted transcriptional and epigenetic regulation in plants.
Arabidopsis
/ genetics
Arabidopsis Proteins
/ genetics
CRISPR-Cas Systems
/ genetics
Epigenesis, Genetic
/ genetics
Feasibility Studies
Gene Expression Regulation, Plant
Genetic Engineering
/ methods
Genetic Vectors
/ genetics
Histone Code
/ genetics
Plants, Genetically Modified
/ genetics
Promoter Regions, Genetic
/ genetics
Transcriptional Activation
Journal
PloS one
ISSN: 1932-6203
Titre abrégé: PLoS One
Pays: United States
ID NLM: 101285081
Informations de publication
Date de publication:
2019
2019
Historique:
received:
19
07
2019
accepted:
23
08
2019
entrez:
27
9
2019
pubmed:
27
9
2019
medline:
3
4
2020
Statut:
epublish
Résumé
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.
Identifiants
pubmed: 31557222
doi: 10.1371/journal.pone.0222778
pii: PONE-D-19-20357
pmc: PMC6762090
doi:
Substances chimiques
Arabidopsis Proteins
0
FT protein, Arabidopsis
0
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Validation Study
Langues
eng
Sous-ensembles de citation
IM
Pagination
e0222778Déclaration de conflit d'intérêts
The authors have declared that no competing interests exist.
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