Specific and heritable genetic interference by double-stranded RNA in <i>Arabidopsis thaliana</i>
Proceedings of the National Academy of SciencesPublished 18 April 2000
Chiou‐Fen Chuang, Elliot M. Meyerowitz
Citations645
SJR quartileQ1
SJR score3.41
SNIP2.38
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TL;DR
In situ hybridization revealed a correlation between a declining AG mRNA accumulation and increasingly severe phenotypes in AG (RNAi) mutants, suggesting that endogenous mRNA is the target of double-stranded RNA-mediated genetic interference.
Abstract
We investigated the potential of double-stranded RNA interference (RNAi) with gene activity in Arabidopsis thaliana. To construct transformation vectors that produce RNAs capable of duplex formation, gene-specific sequences in the sense and antisense orientations were linked and placed under the control of a strong viral promoter. When introduced into the genome of A. thaliana by Agrobacterium-mediated transformation, double-stranded RNA-expressing constructs corresponding to four genes, AGAMOUS (AG), CLAVATA3, APETALA1, and PERIANTHIA, caused specific and heritable genetic interference. The severity of phenotypes varied between transgenic lines. In situ hybridization revealed a correlation between a declining AG mRNA accumulation and increasingly severe phenotypes in AG (RNAi) mutants, suggesting that endogenous mRNA is the target of double-stranded RNA-mediated genetic interference. The ability to generate stably heritable RNAi and the resultant specific phenotypes allows us to selectively reduce gene function in A. thaliana.
Keywords
Agricultural and Biological SciencesBiochemistry, Genetics and Molecular Biology
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The Plant CellEarly flower development in Arabidopsis.
2,090 Citations1990David Smyth, John P. Bowman +1 more
The early development of the flower of Arabidopsis thaliana is described from initiation until the opening of the bud, and the morphogenesis, growth rate, and surface structure of floral organs were recorded in detail using scanning electron microscopy.
NatureThe protein encoded by the Arabidopsis homeotic gene agamous resembles transcription factors
1,631 Citations1990Martin F. Yanofsky, Hong Mā +4 more
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ScienceSignaling of Cell Fate Decisions by <i>CLAVATA3</i> in <i>Arabidopsis</i> Shoot Meristems
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CellThe rde-1 Gene, RNA Interference, and Transposon Silencing in C. elegans
1,308 Citations1999Hiroaki Tabara, Madathia Sarkissian +6 more
It is shown that rde-1 is a member of the piwi/sting/argonaute/zwille/eIF2C gene family conserved from plants to vertebrates and the possibility that one natural function of RNAi is transposon silencing is discussed.
The Plant CellGenes directing flower development in Arabidopsis.
1,219 Citations1989John L. Bowman, David Smyth +1 more
It seems that the wild-type alleles of these four genes allow cells to determine their place in the developing flower and thus to differentiate appropriately and is proposed that these genes may be involved in setting up or responding to concentric, overlapping fields within the flower primordium.
NatureMolecular characterization of the Arabidopsis floral homeotic gene APETALA1
1,160 Citations1992M. Alejandra Mandel, Cindy Gustafson‐Brown +2 more
Cloned the APETALA1 gene is cloned and it is shown that it encodes a putative transcription factor that contains a MADS-domain2 that is uniformly expressed in young flower primordia, and later becomes localized to sepals and petals.
Proceedings of the National Academy of SciencesVirus resistance and gene silencing in plants can be induced by simultaneous expression of sense and antisense RNA
1,132 Citations1998Peter M. Waterhouse, Michael W. Graham +1 more
It is shown that transforming plants with virus or reporter gene constructs that produce RNAs capable of duplex formation confer virus immunity or gene silencing on the plants.
CellUse of dsRNA-Mediated Genetic Interference to Demonstrate that frizzled and frizzled 2 Act in the Wingless Pathway
1,120 Citations1998Jason R. Kennerdell, Richard W. Carthew
Investigation of the potential of double-stranded RNA to interfere with the function of genes in Drosophila demonstrated that dsRNA interference can be used to analyze many aspects of gene function.
DevelopmentControl of flower development in <i>Arabidopsis thaliana</i> by <i>APETALA1</i> and interacting genes
762 Citations1993John L. Bowman, John Paul Alvarez +3 more
The results suggest that the products of APETALA1 and another gene, LEAFY, are required to ensure that primordia arising on the flanks of the inflorescence apex adopt a floral fate, as opposed to becoming an inflorescence shoot.
Cellmut-7 of C. elegans, Required for Transposon Silencing and RNA Interference, Is a Homolog of Werner Syndrome Helicase and RNaseD
746 Citations1999René F. Ketting, Thomas Haverkamp +2 more
This work mutagenized one silenced strain of C. elegans and isolated mutants in which Tc1 had been activated in the germline, finding one of the mutated genes, mut-7, to encode a protein with homology to RNaseD, providing support for the notion that RNAi works by dsRNA-directed, enzymatic RNA degradation.
CellPIF3, a Phytochrome-Interacting Factor Necessary for Normal Photoinduced Signal Transduction, Is a Novel Basic Helix-Loop-Helix Protein
732 Citations1998Min Ni, James M. Tepperman +1 more
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Development<i>CLAVATA3</i> is a specific regulator of shoot and floral meristem development affecting the same processes as <i>CLAVATA1</i>
718 Citations1995Steven E. Clark, Mark Running +1 more
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Proceedings of the National Academy of SciencesDouble-stranded RNA induces mRNA degradation in <i>Trypanosoma</i> <i>brucei</i>
715 Citations1998Huân M. Ngô, Christian Tschudi +2 more
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CellSystemic Spread of Sequence-Specific Transgene RNA Degradation in Plants Is Initiated by Localized Introduction of Ectopic Promoterless DNA
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CellNegative regulation of the Arabidopsis homeotic gene AGAMOUS by the APETALA2 product
674 Citations1991G. Drews, John L. Bowman +1 more
Observations show that the expression patterns of the Arabidopsis floral homeotic genes are in part established by regulatory interactions between these genes.
ScienceRNAi in <i>C. elegans</i> : Soaking in the Genome Sequence
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Double-stranded RNA has recently been shown to trigger sequence-specific gene silencing in a wide variety of organisms, including nematodes, plants, trypanosomes, fruit flies and planaria, and these observations are spurring new inquiries to understand RNA-triggered genetic-control mechanisms and their biological roles.
The Plant CellFunction of the apetala-1 gene during Arabidopsis floral development.
600 Citations1990Vivian F. Irish, I. M. Sussex
It is proposed that the AP1 and the apetala 2 (AP2) genes may encode similar functions that are required to define the pattern of where floral organs arise, as well as for determinate development of the floral meristem.
Proceedings of the National Academy of SciencesRNA as a target of double-stranded RNA-mediated genetic interference in <i>Caenorhabditis elegans</i>
571 Citations1998Mary K. Montgomery, Siqun Xu +1 more
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Proceedings of the National Academy of SciencesA chloroplast lipoxygenase is required for wound-induced jasmonic acid accumulation in Arabidopsis.
571 Citations1995Erin Bell, Robert A. Creelman +1 more
The characterization of LOX2 indicates that the protein is targeted to chloroplasts, and suggests that wound-induced JA (or some other LOx2-requiring component of the wound response pathway) is involved in the wound- induced regulation of this gene.
Proceedings of the National Academy of SciencesDouble-stranded RNA specifically disrupts gene expression during planarian regeneration
544 Citations1999Alejandro Sánchez Alvarado, Phillip A. Newmark
The ability to eliminate gene function in a regenerating organism such as the planarian overcomes previous experimental limitations and opens the study of animal regeneration to unprecedented levels of molecular detail.
NatureRole of SUPERMAN in maintaining Arabidopsis floral whorl boundaries
438 Citations1995Hajime Sakai, Leonard Medrano +1 more
Genes & DevelopmentThe interface of sigma with core RNA polymerase is extensive, conserved, and functionally specialized
417 Citations1999Meghan M. Sharp, Clement L. K. Chan +7 more
The regions of sigma identified perform specialized functions, suggesting that different portions of the interface perform discrete roles during transcription initiation, and that sigma(70) family members use homologous residues, at least in part, to interact with core.
The Plant CellCloning and Functional Characterization of an Arabidopsis Nitrate Transporter Gene That Encodes a Constitutive Component of Low-Affinity Uptake
410 Citations1999Nien‐Chen Huang, Kun-Hsiang Liu +2 more
Cl cloning and characterization of a CHL1 homolog, AtNRT1:2 (originally named NTL1), are reported, with data to indicate that this gene encodes a constitutive component of low-affinity nitrate uptake.
Development<i>CRABS</i> CLAW and <i>SPATULA</i>, two <i>Arabidopsis</i> genes that control carpel development in parallel with <i>AGAMOUS</i>
385 Citations1999John Paul Alvarez, David Smyth
Double mutants of crabs claw and spatula with homeotic mutants that develop ectopic carpels demonstrate that CRABS CLAW and SPATULA are necessary for, and inseparable from, carpel development, and that their action is negatively regulated by A and B organ identity genes.
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Trends in Plant ScienceListening to the silent genes: transgene silencing, gene regulation and pathogen control
366 Citations1999Jan M. Kooter
Plant Molecular BiologyImproved binary vectors for Agrobacterium-mediated plant transformation
341 Citations1990Kevin E. McBride, Kristin R. Summerfelt
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Trends in GeneticsDouble-stranded RNA as a mediator in sequence-specific genetic silencing and co-suppression
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Genes & DevelopmentThe PERIANTHIA gene encodes a bZIP protein involved in the determination of floral organ number in Arabidopsis thaliana
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PLANT PHYSIOLOGYProtective Function of Chloroplast 2-Cysteine Peroxiredoxin in Photosynthesis. Evidence from Transgenic Arabidopsis1
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The Plant JournalSequence‐based identification of T‐DNA insertion mutations in <i>Arabidopsis:</i> actin mutants <i>act2‐1</i> and <i>act4‐1</i>
212 Citations1995Elizabeth C. McKinney, Nazeem Ali +5 more
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183 Citations1997Sherry A. Kempin, Sarah J. Liljegren +4 more
This work has successfully disrupted the AGL5 MADS-box gene in Arabidopsis by homologous recombination, providing a useful tool for future analyses.
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180 Citations1998Yang Shi, C. Mello
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158 Citations1996Mark Running, Elliot M. Meyerowitz
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It is shown that co-suppression and other types of post-transcriptional gene silencing in plants are initiated by homology-dependent pairing of transgene sequences, and the silencing mechanism acts in the cytoplasm to prevent accumulation of all RNAs with homology to thetransgene.
Arabidopsis
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54 Citations1999Helle Naver, Anna Haldrup +1 more
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