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dc.contributor.author
Jay, Florence
dc.contributor.author
Vitel, Maxime
dc.contributor.author
Brioudes, Florian
dc.contributor.author
Louis, Mélissa
dc.contributor.author
Knobloch, Thomas
dc.contributor.author
Voinnet, Olivier
dc.date.accessioned
2019-08-30T12:48:11Z
dc.date.available
2019-06-06T12:14:15Z
dc.date.available
2019-06-06T12:37:16Z
dc.date.available
2019-08-30T12:48:11Z
dc.date.issued
2019-09
dc.identifier.issn
1355-8382
dc.identifier.issn
1469-9001
dc.identifier.other
10.1261/rna.068627.118
en_US
dc.identifier.uri
http://hdl.handle.net/20.500.11850/345960
dc.identifier.doi
10.3929/ethz-b-000345960
dc.description.abstract
RNAi mediated by small-interfering RNAs (siRNAs) operates via transcriptional (TGS) and posttranscriptional gene silencing (PTGS). In Arabidopsis thaliana, TGS relies on DICER-LIKE-3 (DCL3)-dependent 24-nt siRNAs loaded into AGO4-clade ARGONAUTE effector proteins. PTGS operates via DCL4-dependent 21-nt siRNAs loaded into AGO1-clade proteins. We set up and validated a medium-throughput, semi-automatized procedure enabling chemical screening, in a 96-well in vitro format, of Arabidopsis transgenic seedlings expressing an inverted-repeat construct from the phloem companion cells. The ensuing quantitative PTGS phenotype was exploited to identify molecules, which, upon topical application, either inhibit or enhance siRNA biogenesis/activities. The vast majority of identified modifiers were enhancers, among which Sortin1, Isoxazolone, and [5-(3,4-dichlorophenyl)furan-2-yl]-piperidine-1-ylmethanethione (DFPM) provided the most robust and consistent results, including upon their application onto soil-grown plants in which their effect was nonautonomous and long lasting. The three molecules increased the RNAi potency of the inverted-repeat construct, in large part by enhancing 21-nt siRNA accumulation and loading into AGO1, and concomitantly reducing AGO4 and DCL3 levels in planta. A similar, albeit not identical effect, was observed on 22-nt siRNAs produced from a naturally occurring inverted-repeat locus, demonstrating that the molecules also enhance endogenous PTGS. In standardized assays conducted in seedling extracts, the three enhancers selectively increased DCL4-mediated processing of in vitro-synthesized double-stranded RNAs, indicating the targeting of a hitherto unknown PTGS component probably independent of the DCL4-cofactor DOUBLE-STRANDED RNA-BINDING 4 (DRB4). This study establishes the proof-of-concept that RNAi efficacy can be modulated by chemicals in a whole organism. Their potential applications and the associated future research are discussed.
en_US
dc.format
application/pdf
en_US
dc.language.iso
en
en_US
dc.publisher
Cold Spring Harbor Laboratory
en_US
dc.subject
Arabidopsis
en_US
dc.subject
Chemical screen
en_US
dc.subject
enhancers-suppressors
en_US
dc.subject
in vitro assay
en_US
dc.subject
RNAi
en_US
dc.subject
siRNA
en_US
dc.title
Chemical enhancers of post-transcriptional gene-silencing in Arabidopsis
en_US
dc.type
Journal Article
dc.date.published
2019-06-04
ethz.journal.title
RNA
ethz.journal.volume
25
en_US
ethz.journal.issue
9
en_US
ethz.pages.start
1078
en_US
ethz.pages.end
1090
en_US
ethz.version.deposit
publishedVersion
en_US
ethz.grant
High resolution and chemical genetic approaches to RNA silencing mechanisms
en_US
ethz.identifier.wos
ethz.identifier.scopus
ethz.publication.place
Cold Spring Harbor, NY
en_US
ethz.publication.status
published
en_US
ethz.leitzahl
ETH Zürich::00002 - ETH Zürich::00012 - Lehre und Forschung::00007 - Departemente::02030 - Dep. Biologie / Dep. of Biology::02541 - Institut für Molekulare Pflanzenbiologie / Institute of Molecular Plant Biology::03876 - Voinnet, Olivier / Voinnet, Olivier
en_US
ethz.leitzahl.certified
ETH Zürich::00002 - ETH Zürich::00012 - Lehre und Forschung::00007 - Departemente::02030 - Dep. Biologie / Dep. of Biology::02541 - Institut für Molekulare Pflanzenbiologie / Institute of Molecular Plant Biology::03876 - Voinnet, Olivier / Voinnet, Olivier
en_US
ethz.grant.agreementno
323071
ethz.grant.fundername
EC
ethz.grant.funderDoi
10.13039/501100000780
ethz.grant.program
FP7
ethz.date.deposited
2019-06-06T12:14:22Z
ethz.source
FORM
ethz.eth
yes
en_US
ethz.availability
Embargoed
en_US
ethz.date.embargoend
2020-10-15
ethz.rosetta.installDate
2019-08-30T12:48:27Z
ethz.rosetta.lastUpdated
2019-08-30T12:48:27Z
ethz.rosetta.exportRequired
true
ethz.rosetta.versionExported
true
ethz.COinS
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