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

Chuang, C.F., Meyerowitz, E.M.
Proceedings of the National Academy of Sciences of the United States of America 2000 v.97 no.9 pp. 4985-4990
Arabidopsis thaliana, RNA, molecular conformation, genetic transformation, Agrobacterium tumefaciens, gene expression, messenger RNA, genes, transgenic plants, reporter genes, beta-glucuronidase, inheritance (genetics)
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.