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PMID: 10781109 Published · ppublish English Journal Article Research Support, U.S. Gov't, P.H.S.

Specific and heritable genetic interference by double-stranded RNA in Arabidopsis thaliana.

Proceedings of the National Academy of Sciences of the United States of America ·Vol. 97 ·No. 9 ·2000-04-25 ·Pages 4985-90

Chuang CF, Meyerowitz EM

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.

MeSH Terms
AGAMOUS Protein, Arabidopsis Arabidopsis/genetics Arabidopsis Proteins DNA, Plant/genetics DNA-Binding Proteins/genetics Homeodomain Proteins/genetics In Situ Hybridization MADS Domain Proteins Plant Proteins/genetics Plants, Genetically Modified RNA, Double-Stranded/genetics RNA, Messenger/genetics Rhizobium/genetics Transcription Factors/genetics Transformation, Genetic
Chemicals
AGAMOUS Protein, Arabidopsis AP1 protein, Arabidopsis AT2G27250 protein, Arabidopsis Arabidopsis Proteins DNA, Plant DNA-Binding Proteins Homeodomain Proteins MADS Domain Proteins PAN protein, Arabidopsis Plant Proteins RNA, Double-Stranded RNA, Messenger Transcription Factors
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Chuang C F
Division of Biology 156-29, California Institute of Technology, Pasadena, CA 91125, USA.
Meyerowitz E M
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Article Info
Journal
Proceedings of the National Academy of Sciences of the United States of America
Abbr.
Proc Natl Acad Sci U S A
ISSN
0027-8424
DOI
10.1073/pnas.060034297
Published
2000-04-25
Pages
4985-90
Language
English
Region
United States
NLM ID
7505876
PMCID
PMC18344
Subset
IM
Grants
NIGMS NIH HHS · R01 GM045697 · United States
NIGMS NIH HHS · GM45697 · United States
External Links
PubMed source DOI: 10.1073/pnas.060034297 PMC Full Text View journal details
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