Efficient silencing of endogenous microRNAs using artificial microRNAs in Arabidopsis thaliana

Eamens, Andrew L., Agius, Claire, Smith, Neil A., Waterhouse, Peter M., & Wang, Ming-Bo (2011) Efficient silencing of endogenous microRNAs using artificial microRNAs in Arabidopsis thaliana. Molecular Plant, 4(1), pp. 157-170.

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Abstract

We report here that the expression of endogenous microRNAs (miRNAs) can be efficiently silenced in Arabidopsis thaliana (Arabidopsis) using artificial miRNA (amiRNA) technology. We demonstrate that an amiRNA designed to target a mature miRNA directs silencing against all miRNA family members, whereas an amiRNA designed to target the stem-loop region of a miRNA precursor transcript directs silencing against only the individual family member targeted. Furthermore, our results indicate that amiRNAs targeting both the mature miRNA and stem-loop sequence direct RNA silencing through cleavage of the miRNA precursor transcript, which presumably occurs in the nucleus of a plant cell during the initial stages of miRNA biogenesis. This suggests that small RNA (sRNA)-guided RNA cleavage in plants occurs not only in the cytoplasm, but also in the nucleus. Many plant miRNA gene families have been identified via sequencing and bioinformatic analysis, but, to date, only a small tranche of these have been functionally characterized due to a lack of effective forward or reverse genetic tools. Our findings therefore provide a new and powerful reverse-genetic tool for the analysis of miRNA function in plants. © The Author 2010. Published by the Molecular Plant Shanghai Editorial Office in association with Oxford University Press on behalf of CSPP and IPPE, SIBS, CAS.

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ID Code: 65914
Item Type: Journal Article
Refereed: Yes
Additional URLs:
Keywords: Artificial microRNA, MicroRNA, RNA silencing, plant RNA, Arabidopsis, article, chemistry, conformation, gene expression regulation, gene silencing, genetics, metabolism, molecular genetics, nucleotide sequence, synthesis, Base Sequence, Gene Expression Regulation, Plant, MicroRNAs, Molecular Sequence Data, Nucleic Acid Conformation, RNA, Plant, Arabidopsis thaliana
DOI: 10.1093/mp/ssq061
ISSN: 1674-2052
Subjects: Australian and New Zealand Standard Research Classification > BIOLOGICAL SCIENCES (060000) > GENETICS (060400)
Australian and New Zealand Standard Research Classification > BIOLOGICAL SCIENCES (060000) > PLANT BIOLOGY (060700)
Divisions: Current > Schools > School of Earth, Environmental & Biological Sciences
Current > QUT Faculties and Divisions > Science & Engineering Faculty
Copyright Owner: Copyright 2010 The Author
Copyright Statement: Published by the Molecular Plant Shanghai Editorial
Office in association with Oxford University Press on behalf of CSPP and
IPPE, SIBS, CAS.
Deposited On: 09 Jan 2014 02:28
Last Modified: 05 Feb 2015 03:25

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