Reverse genetics in the mosquito Anopheles gambiae:: targeted disruption of the Defensin gene
Reverse genetics in the mosquito Anopheles gambiae:: targeted disruption of the Defensin gene
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DOI：
10.1093/embo-reports/kvf180
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发表时间：
2002-09-01
期刊：
影响因子：
7.7
通讯作者：
Levashina, EA
中科院分区：
文献类型：
--
作者：
Blandin, S;Moita, LF;Levashina, EA
冈比亚按蚊是人类疟疾的主要传播媒介，是研究媒介-寄生虫相互作用的重要模式昆虫。在此，我们开发了一种简单的体内双链RNA（dsRNA）敲除方法来确定蚊子抗菌肽基因Defensin的功能。我们将dsRNA注射到成年人中，并观察到防御素的有效和可重复的沉默。敲低表型的分析显示，这种肽是蚊子对革兰氏阳性菌的抗菌防御所必需的。相比之下，在感染伯氏疟原虫的蚊子中，在不存在防御素的情况下，没有观察到蚊子生存力的损失，也没有观察到对寄生虫中肠阶段的发育和形态的显著影响。我们的结论是，该肽不是一个主要的抗寄生虫因子在A。冈比亚体内。本研究结果为蚊子体内基因功能的研究开辟了新的视角，并为通过靶向基因沉默进行基因组规模的系统功能筛选奠定了基础。
Anopheles gambiae, the major vector of human malaria parasite, is an important insect model to study vector-parasite interactions. Here, we developed a simple in vivo double-stranded RNA (dsRNA) knockout approach to determine the function of the mosquito antimicrobial peptide gene Defensin. We injected dsRNA into adults and observed efficient and reproducible silencing of Defensin. Analysis of the knockdown phenotype revealed that this peptide is required for the mosquito antimicrobial defense against Gram-positive bacteria. In contrast, in mosquitoes infected by Plasmodium berghei, no loss of mosquito viability and no significant effect on the development and morphology of the parasite midgut stages were observed in the absence of Defensin. We conclude that this peptide is not a major antiparasitic factor in A. gambiae in vivo. Our results open new perspectives for the study of mosquito gene function in vivo and provide a basis for genome-scale systematic functional screens by targeted gene silencing.