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Proc Natl Acad Sci U S A. 2000 Apr 25;97(9):4985-90.

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

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1
Division of Biology 156-29, California Institute of Technology, Pasadena, CA 91125, USA.

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.

PMID:
10781109
PMCID:
PMC18344
DOI:
10.1073/pnas.060034297
[Indexed for MEDLINE]
Free PMC Article
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