Transgenic alteration of Toll immune pathway in the female mosquito Aedes aegypti
Transgenic alteration of Toll immune pathway in the female mosquito Aedes aegypti
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DOI：
10.1073/pnas.0502815102
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发表时间：
2005-09-20
影响因子：
11.1
通讯作者：
Raikhel, AS
中科院分区：
文献类型：
--
作者：
Bian, G;Shin, SW;Raikhel, AS
反向遗传学是了解蚊子先天免疫中基因功能及其相互作用的有力工具。我们采用转基因技术结合RNA干扰（RNAi）技术，研究了果蝇（Drosophila Dorsal）同源基因mosquito 1在埃及伊蚊（Aedes aegypti）Toll免疫途径中的调控作用。通过用pBac[3xP 3-EGFP，afm]载体用Delta Aa 11-A或由雌性脂肪体特异性卵黄蛋白原（Vg）启动子驱动的Aa 11的双链RNA构建体转化蚊子，我们产生了两种不同的转基因蚊子品系，一种具有过表达的Aa 11，第二种具有Aa 11敲低。两种品系都具有各自转基因的单拷贝，并且在两种转基因蚊子中的表达通过血液喂养被高度激活。Vg-Δ Toll 1-A转基因蚊子通过吸血激活脂肪体中的Toll免疫途径。在Vg-Δ A1-A转基因蚊子中，Aaa 1-A和Aaa 1-B两种亚型的过表达导致了与脓毒性损伤无关的斑蚊1A和Serpin-27 A的伴随激活。在RNAi敲低果蝇仙人掌的伊蚊同源物（果蝇Toll途径的I κ B抑制剂）的蚊子中观察到相同的表型。转基因RNAi敲除Aaa 11对蚊子先天免疫的影响通过增加对昆虫病原真菌球孢白僵菌的易感性和减少真菌攻击后Spz 1A和Serpin-27 A基因表达的诱导来揭示。这些结果证明了AaB 1是蚊Toll免疫途径的关键下游调节因子。埃及人。
Reverse genetics is a powerful tool for understanding gene functions and their interactions in the mosquito innate immunity. We took the transgenic approach, in combination with the RNA interference (RNAi) technique, to elucidate the role of mosquito REL1, a homolog of Drosophila Dorsal, in regulation of Toll immune pathway in the mosquito Aedes aegypti. By transforming the mosquitoes with Delta REL1-A or a double-stranded RNA construct of REL1 driven by the female fat body-specific vitellogenin (Vg) promoter with the pBac[3xP3-EGFP, afm] vector, we generated two different transgenic mosquito strains, one with overexpressed AaREL1 and the second with AaREL1 knockdown. Both strains had a single copy of the respective transgene, and the expression in both transgenic mosquitoes was highly activated by blood feeding. Vg-Delta REL1-A transgenic mosquitoes activate Toll immune pathway in the fat body by blood feeding. The overexpression of both isoforms, AaREL1-A and AaREL1-B, in Vg-Delta REL1-A transgenic mosquitoes resulted in the concomitant activation of Aedes Spatzle1A and Serpin-27A, independent of septic injury. The same phenotype was observed in the mosquitoes with RNAi knockdown of an Aedes homolog to Drosophila cactus, an I kappa B inhibitor of Drosophila Toll pathway. The effect of the transgenic RNAi knockdown of AaREL1 on mosquito innate immunity was revealed by increased susceptibility to the entomopathogenic fungus Beauveria bassiana and the reduced induction of Spz1A and Serpin-27A gene expression after fungal challenge. These results have proven that AaREL1 is a key downstream regulator of Toll immune pathway in the mosquito A. aegypti.