Specific and heritable genetic interference by double-stranded RNA in <i>Arabidopsis thaliana</i>

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  • Chiou-Fen Chuang
    Division of Biology 156–29, California Institute of Technology, Pasadena, CA 91125
  • Elliot M. Meyerowitz
    Division of Biology 156–29, California Institute of Technology, Pasadena, CA 91125

抄録

<jats:p> We investigated the potential of double-stranded RNA interference (RNAi) with gene activity in <jats:italic>Arabidopsis thaliana</jats:italic> . 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 <jats:italic>A. thaliana</jats:italic> by <jats:italic>Agrobacterium</jats:italic> -mediated transformation, double-stranded RNA-expressing constructs corresponding to four genes, <jats:italic>AGAMOUS</jats:italic> ( <jats:italic>AG</jats:italic> ), <jats:italic>CLAVATA3</jats:italic> , <jats:italic>APETALA1</jats:italic> , and <jats:italic>PERIANTHIA</jats:italic> , caused specific and heritable genetic interference. The severity of phenotypes varied between transgenic lines. <jats:italic>In situ</jats:italic> hybridization revealed a correlation between a declining <jats:italic>AG</jats:italic> mRNA accumulation and increasingly severe phenotypes in <jats:italic>AG</jats:italic> ( <jats:italic>RNAi</jats:italic> ) 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 <jats:italic>A. thaliana</jats:italic> . </jats:p>

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