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Published April 25, 2000 | Published
Journal Article Open

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

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.

Additional Information

© 2000 National Academy of Sciences. Contributed by Elliot M. Meyerowitz, January 24, 2000. Published online before print April 18, 2000. We thank John Bowman and Yuval Eshed for critical reading of the manuscript; Xavier Ambroggio, Catherine Baker, Chieh Chang, Toshiro Ito, Jeff Long, Kazuaki Ohashi, Carolyn Ohno, G. Venugopala Reddy, Doris Wagner, Frank Wellmer, and Eva Ziegelhoffer for their comments on the manuscript; Amani Zewail for technical assistance; and Toshiro Ito for providing the anti-AG antibody. This work was supported by National Institutes of Health Grant GM45697 to E.M.M.

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