Specific and heritable genetic interference by double-stranded RNA in Arabidopsis thaliana.
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.
Chuang
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CF
Division of Biology 156-29, California Institute of Technology, Pasadena, CA 91125, USA.
Meyerowitz
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EM
D026342:AGAMOUS Protein, Arabidopsis; D017360:Arabidopsis / Q000235:genetics*; D029681:Arabidopsis Proteins; D018744:DNA, Plant / Q000235:genetics; D004268:DNA-Binding Proteins / Q000235:genetics; D018398:Homeodomain Proteins / Q000235:genetics; D017403:In Situ Hybridization; D026161:MADS Domain Proteins; D010940:Plant Proteins / Q000235:genetics; D030821:Plants, Genetically Modified; D012330:RNA, Double-Stranded / Q000235:genetics*; D012333:RNA, Messenger / Q000235:genetics; D012231:Rhizobium / Q000235:genetics; D014157:Transcription Factors / Q000235:genetics; D014170:Transformation, Genetic
D026342:AGAMOUS Protein, Arabidopsis; C091535:AP1 protein, Arabidopsis; C118167:AT2G27250 protein, Arabidopsis; D029681:Arabidopsis Proteins; D018744:DNA, Plant; D004268:DNA-Binding Proteins; D018398:Homeodomain Proteins; D026161:MADS Domain Proteins; C118078:PAN protein, Arabidopsis; D010940:Plant Proteins; D012330:RNA, Double-Stranded; D012333:RNA, Messenger; D014157:Transcription Factors