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Heritable gene silencing in Drosophila using double-stranded RNA

Nature Biotechnology volume 18, pages 896–898 (2000)Cite this article

Abstract

RNA-mediated interference (RNAi) is a recently discovered method to determine gene function in a number of organisms, including plants1, nematodes2, Drosophila3,4, zebrafish5, and mice6. Injection of double-stranded RNA (dsRNA) corresponding to a single gene into organisms silences expression of the specific gene2,3,4,5,6. Rapid degradation of mRNA in affected cells blocks gene expression2. Despite the promise of RNAi as a tool for functional genomics, injection of dsRNA interferes with gene expression transiently and is not stably inherited2,3,5,6. Consequently, use of RNAi to study gene function in the late stages of development has been limited. It is particularly problematic for development of disease models that reply on post-natal individuals. To circumvent this problem in Drosophila, we have developed a method to express dsRNA as an extended hairpin-loop RNA. This method has recently been successful in generating RNAi in the nematode Caenorhabditis elegans7. The hairpin RNA is expressed from a transgene exhibiting dyad symmetry in a controlled temporal and spatial pattern. We report that the stably inherited transgene confers specific interference of gene expression in embryos, and tissues that give rise to adult structures such as the wings, legs, eyes, and brain. Thus, RNAi can be adapted to study late-acting gene function in Drosophila. The success of this approach in Drosophila and C. elegans suggests that a similar approach may prove useful to study gene function in higher organisms for which transgenic technology is available.

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Figure 1: Effect of injection of hairpin-loop RNA on Drosophila embryos.
Figure 2: Effect of transgenic hairpin-loop RNA on target gene expression.

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  1. Department of Biological Sciences, University of Pittsburgh, 4249 Fifth Avenue, 217 Clapp Hall, Pittsburgh, 15260, PA

    Jason R. Kennerdell & Richard W. Carthew

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  1. Jason R. Kennerdell
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  2. Richard W. Carthew
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Correspondence to Richard W. Carthew.

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Kennerdell, J., Carthew, R. Heritable gene silencing in Drosophila using double-stranded RNA. Nat Biotechnol 18, 896–898 (2000). https://doi.org/10.1038/78531

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