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Highly specific gene silencing by artificial miRNAs in rice

Chen, Hao; Warthmann, Norman; Ossowski, Stephan; Weigel, Detlef; Hervé, Philippe

Description

BACKGROUND Endogenous microRNAs (miRNAs) are potent negative regulators of gene expression in plants and animals. Artificial miRNAs (amiRNAs)-designed to target one or several genes of interest-provide a new and highly specific approach for effective post-transcriptional gene silencing (PTGS) in plants. METHODOLOGY We devised an amiRNA-based strategy for both japonica and indica type strains of cultivated rice, Oryza sativa. Using an endogenous rice miRNA precursor and customized 21mers, we...[Show more]

dc.contributor.authorChen, Hao
dc.contributor.authorWarthmann, Norman
dc.contributor.authorOssowski, Stephan
dc.contributor.authorWeigel, Detlef
dc.contributor.authorHervé, Philippe
dc.date.accessioned2015-11-16T00:49:16Z
dc.date.available2015-11-16T00:49:16Z
dc.identifier.issn1932-6203
dc.identifier.urihttp://hdl.handle.net/1885/16491
dc.description.abstractBACKGROUND Endogenous microRNAs (miRNAs) are potent negative regulators of gene expression in plants and animals. Artificial miRNAs (amiRNAs)-designed to target one or several genes of interest-provide a new and highly specific approach for effective post-transcriptional gene silencing (PTGS) in plants. METHODOLOGY We devised an amiRNA-based strategy for both japonica and indica type strains of cultivated rice, Oryza sativa. Using an endogenous rice miRNA precursor and customized 21mers, we designed amiRNA constructs targeting three different genes (Pds, Spl11, and Eui1/CYP714D1). Upon constitutive expression of these amiRNAs in the varieties Nipponbare (japonica) and IR64 (indica), the targeted genes are down-regulated by amiRNA-guided cleavage of the transcripts, resulting in the expected mutant phenotypes. The effects are highly specific to the target gene, the transgenes are stably inherited and they remain effective in the progeny. CONCLUSION/SIGNIFICANCE Our results not only show that amiRNAs can efficiently trigger gene silencing in a monocot crop, but also that amiRNAs can effectively modulate agronomically important traits in varieties used in modern breeding programs. We provide all software tools and a protocol for the design of rice amiRNA constructs, which can be easily adapted to other crops. The approach is suited for candidate gene validation, comparative functional genomics between different varieties, and for improvement of agronomic performance and nutritional value.
dc.description.sponsorshipThis work was supported by the Max Planck Society, of which D.W. is a director, by European Community FP6 IP SIROCCO (contract LSHG-CT-2006- 037900), and by the USAID-funded project entitled Development of rice biotechnology products for Asia: technical and pre-regulatory components at the International Rice Research Institute (IRRI) coordinated by P.H. and Gerard Barry.
dc.format10 pages
dc.publisherPublic Library of Science
dc.rights© 2008 Warthmann et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
dc.sourcePLoS ONE
dc.subjectgenes, plant
dc.subjectmicrornas
dc.subjectnucleic acid conformation
dc.subjectoryza sativa
dc.subjectreverse transcriptase polymerase chain reaction
dc.subjectgene silencing
dc.titleHighly specific gene silencing by artificial miRNAs in rice
dc.typeJournal article
local.description.notesImported from ARIES. At the time of publication, Norman Warthmann was affiliated with Department of Molecular Biology, Max Planck Institute for Developmental Biology, Tubingen, Germany.
local.identifier.citationvolume3
dcterms.dateAccepted2008-02-19
dc.date.issued2008-03-19
local.identifier.absfor060102
local.identifier.absfor060702
local.identifier.absfor060113
local.identifier.ariespublicationU3488905xPUB1097
local.publisher.urlhttps://www.plos.org/
local.type.statusPublished Version
local.contributor.affiliationWarthmann, Norman, College of Medicine, Biology and Environment, CMBE Research School of Biology, Division of Plant Sciences, The Australian National University
local.contributor.affiliationChen, Hao, International Rice Research Institute (IRRI), Philippines
local.contributor.affiliationOssowski, Stephan, Max Planck Institute for Developmental Biology, Germany
local.contributor.affiliationWeigel, Detlef, Max Planck Institute for Developmental Biology,, Germany
local.contributor.affiliationHerve, Philippe, International Rice Research Institute (IRRI), Philippines
local.identifier.essn1932-6203
local.bibliographicCitation.issue3
local.bibliographicCitation.startpagee1829
local.bibliographicCitation.lastpage10
local.identifier.doi10.1371/journal.pone.0001829
local.identifier.absseo970106
local.identifier.absseo820402
dc.date.updated2015-12-10T10:02:01Z
local.identifier.scopusID2-s2.0-46649106719
local.identifier.thomsonID000260762300031
CollectionsANU Research Publications

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