Abstract
Double-stranded RNA (dsRNA) gene interference is an efficient method to silence gene expression in a sequence-specific manner. Here we show that the direct injection of dsRNA can be used in adult Drosophila flies to disrupt function of endogenous genes in vivo. As a proof of principle, we have used this method to silence components of a major signaling cascade, the Toll pathway, which controls fruit fly resistance to fungal and Gram-positive bacterial infections. We demonstrate that the knockout is efficient only if dsRNA is injected in 4- or more day-old flies and that it lasts for at least 1 week. Furthermore, we report dsRNA-based epistatic gene analysis via injection of a mixture of two dsRNAs and propose that injection of dsRNA represents a powerful method for rapid functional analysis of genes in Drosophila melanogaster adults, particularly of those whose mutations are lethal during development.
Publication types
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Research Support, Non-U.S. Gov't
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Research Support, U.S. Gov't, P.H.S.
MeSH terms
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Animals
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Drosophila Proteins / genetics
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Drosophila Proteins / metabolism*
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Drosophila melanogaster / drug effects
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Drosophila melanogaster / genetics*
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Drosophila melanogaster / growth & development
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Epistasis, Genetic
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Female
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Green Fluorescent Proteins
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Homeodomain Proteins / genetics
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Luminescent Proteins / genetics
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Phenotype
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RNA Interference*
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RNA, Double-Stranded / administration & dosage
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RNA, Double-Stranded / genetics*
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Receptors, Cell Surface / genetics
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Receptors, Cell Surface / metabolism*
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Serpins / genetics
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Signal Transduction / genetics
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Time Factors
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Toll-Like Receptors
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Transcription Factors / genetics
Substances
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Drosophila Proteins
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Homeodomain Proteins
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Luminescent Proteins
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Opa protein, Drosophila
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RNA, Double-Stranded
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Receptors, Cell Surface
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Serpins
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Tl protein, Drosophila
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Toll-Like Receptors
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Transcription Factors
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nec protein, Drosophila
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Green Fluorescent Proteins