Silencing of Toll pathway components by direct injection of double-stranded RNA into Drosophila adult flies
Silencing of Toll pathway components by direct injection of double-stranded RNA into Drosophila adult flies
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DOI：
10.1093/nar/gkg852
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发表时间：
2003-11-15
影响因子：
14.9
通讯作者：
Levashina, EA
中科院分区：
文献类型：
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作者：
Goto, A;Blandin, S;Levashina, EA
双链RNA（dsRNA）基因干扰是一种有效的序列特异性基因沉默方法。在这里，我们表明，直接注射的dsRNA可以用于成年果蝇在体内破坏内源性基因的功能。作为原理的证明，我们已经使用这种方法来沉默一个主要的信号级联，Toll通路，控制果蝇对真菌和革兰氏阳性细菌感染的抗性的组件。我们证明，敲除是有效的，只有当dsRNA注射在4天或更大的苍蝇，它持续至少1周。此外，我们报告的dsRNA为基础的上位基因分析，通过注射的混合物的两个dsRNA，并提出注射的dsRNA是一个强大的方法，在果蝇成年人的基因的快速功能分析，特别是那些在发展过程中的突变是致命的。
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.