A conditional silencing suppression system for transient expression.

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State: Public
Version: Final published version
Serval ID
serval:BIB_56B986180F40
Type
Article: article from journal or magazin.
Collection
Publications
Institution
Title
A conditional silencing suppression system for transient expression.
Journal
Scientific Reports
Author(s)
Reis R.S., Litholdo C.G., Bally J., Roberts T.H., Waterhouse P.M.
ISSN
2045-2322 (Electronic)
ISSN-L
2045-2322
Publication state
Published
Issued date
2018
Peer-reviewed
Oui
Volume
8
Number
1
Pages
9426
Language
english
Abstract
RNA silencing is a powerful tool deployed by plants against viral infection and abnormal gene expression. Plant viruses have evolved a suite of silencing suppressors for counter-defense, which are also widely used to boost transcript and protein accumulation in transient assays. However, only wild type silencing suppressor proteins have been reported to date. Here we demonstrate that P0 of Potato leafroll virus (PLRV), PLP0, can be split into two proteins that only show silencing suppression activity upon co-expression. We cloned each of these proteins in two different constructs and transiently co-infiltrated them in N. benthamiana leaves. We expressed a fluorescent protein from one of the vectors and observed that cells expressing both halves of PLP0 suppressed gene silencing. Further, we showed that Q system of Neurospora crassa, based on co-expression of a transcription activator and inhibitor, is functional in agroinfiltrated leaves of N. benthamiana. Q system combined with the split PLP0 system showed very tight co-expression of Q system's transcriptional activator and inhibitor. Altogether, our experiments demonstrate a functioning conditional silencing suppressor system and its potential as a powerful tool for transient expression in N. benthamiana leaves, as well as the application of the Q system in plants.
Pubmed
Web of science
Open Access
Yes
Create date
12/07/2018 12:12
Last modification date
20/08/2019 14:10
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