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Published VersionCC BY-NC V4.0, Open Access
Journal article
A 22-nt artificial microRNA mediates widespread RNA silencing in Arabidopsis
The Plant Journal , Vol.76(3), pp.519-529
2013
PMCID: PMC4241025
PMID: 23937661
Abstract
It is known that 22-nucleotide (nt) microRNAs (miRNAs) derived from asymmetric duplexes trigger phased small-interfering RNA (phasiRNA) production from complementary targets. Here we investigate the efficacy of 22-nt artificial miRNA (amiRNA)-mediated RNA silencing relative to conventional hairpin RNA (hpRNA) and 21-nt amiRNA-mediated RNA silencing. CHALCONE SYNTHASE (CHS) was selected as a target in Arabidopsis thaliana due to the obvious and non-lethal loss of anthocyanin accumulation upon widespread RNA silencing. Over-expression of CHS in the pap1-D background facilitated visual detection of both local and systemic RNA silencing. RNA silencing was initiated in leaf tissues from hpRNA and amiRNA plant expression vectors under the control of an Arabidopsis RuBisCo small subunit 1A promoter (SSU). In this system, hpRNA expression triggered CHS silencing in most leaf tissues but not in roots or seed coats. Similarly, 21-nt amiRNA expression from symmetric miRNA/miRNA* duplexes triggered CHS silencing in all leaf tissues but not in roots or seed coats. However, 22-nt amiRNA expression from an asymmetric duplex triggered CHS silencing in all tissues, including roots and seed coats, in the majority of plant lines. This widespread CHS silencing required RNA-DEPENDENT RNA POLYMERASE6-mediated accumulation of phasiRNAs from the endogenous CHS transcript. These results demonstrate the efficacy of asymmetric 22-nt amiRNA-directed RNA silencing and associated phasiRNA production and activity, in mediating widespread RNA silencing of an endogenous target gene. Asymmetric 22-nt amiRNA-directed RNA silencing requires little modification of existing amiRNA technology and is expected to be effective in suppressing other genes and/or members of gene families.
Details
- Title
- A 22-nt artificial microRNA mediates widespread RNA silencing in Arabidopsis
- Authors
- M McHale (Author) - The University of SydneyAndrew Eamens (Author) - The University of SydneyE. Jean Finnegan (Author) - CSIRO Plant Industry (Australia)Peter M. Waterhouse (Author) - The University of Sydney
- Publication details
- The Plant Journal , Vol.76(3), pp.519-529
- Publisher
- Wiley-Blackwell Publishing Ltd.
- Date published
- 2013
- DOI
- 10.1111/tpj.12306
- ISSN
- 1365-313X; 0960-7412
- PMID
- 23937661; PMC4241025
- Copyright note
- © 2013 The Authors. The Plant Journal © 2013 John Wiley & Sons Ltd. This is an open access article under the terms of the Creative Commons Attribution-NonCommercial License,which permits use, distribution and reproduction in any medium, provided the original work is properly cited and is not used for commercial purposes.
- Organisation Unit
- School of Health - Biomedicine; University of the Sunshine Coast, Queensland; School of Health and Behavioural Sciences - Legacy
- Language
- English
- Record Identifier
- 99656590502621
- Output Type
- Journal article
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