Elucidation of mosquito-malaria parasite interaction using genetically engineered mosquitoes

疟原虫子孢子侵入蚊子的唾液腺，并在唾液管中等待下一次吸血。唾液腺侵袭的过程和分子机制仍不清楚。为了在斯氏按蚊中建立一个强大的唾液腺特异性转基因表达，我们获得了一个编码按蚊抗血小板聚集蛋白（AAPP）基因的唾液腺特异性启动子。aapp启动子是女性唾液腺特异性和血餐诱导型强启动子。使用这个启动子，我们产生了一个转基因的斯氏按蚊表达丰富的Discosoma sp.红色荧光蛋白（DsRed）的远侧叶的腺体，其中子孢子入侵优先。这些结果为阐明唾液腺-寄生虫相互作用和产生抗寄生虫转基因蚊子提供了可能性。 ...更多信息 疟疾控制的新战略，并提供了一个强有力的工具，调查蚊子寄生虫的相互作用。我们使用转基因蚊子阻断传播的研究一直集中在寄生虫的中肠阶段，干扰动合子入侵或卵囊分化。我们引入转基因蚊子的分子是从Cucumaria echinata（海洋无脊椎动物）中分离的CEL-III。CEL-III作为凝集素具有很强的溶血活性和血凝活性。CEL-III基因表达由羧肽酶启动子（中肠特异性启动子）驱动。正如预期的那样，在血餐后6小时在中肠中诱导CEL-III表达。我们在同一只感染的大鼠上喂养对照蚊子和转基因蚊子，并测量形成的卵囊数量。转基因蚊子的传播阻断效率为91%，与对照蚊子相比。因此，CEL-III在蚊子中肠中的表达通过抑制疟原虫发育而严重降低了载体能力。由于CEL-III靶向红细胞而不是寄生虫，我们预计这种CEL-III转基因蚊子将能够削弱所有疟原虫物种的寄生虫发育。少

Malaria sporozoites invade the mosquito salivary glands and wait in the salivary duct until the next blood feeding. The mechanisms of the process and molecules involved in the salivary gland invasion remain largely unknown. To establish a robust salivary gland-specific transgene expression in Anopheles stephensi, we obtained a salivary gland-specific promoter for a gene encoding Anopheles anti-platelet aggregation protein (AAPP). The aapp promoter is a female salivary gland-specific and blood meal-inducible strong promoter. Using this promoter, we generated a transgenic An.stephensi expressing abundant Discosoma sp. red fluorescent protein (DsRed) in the distal-lateral lobes of the glands, where the sporozoites invade preferentially. These results open up the possibilities of elucidating salivary gland-parasite interactions and generating transgenic mosquitoes refractory to parasites.Recent advances in the genetic engineering of anopheline mosquitoes have raised hopes for their use as … More new strategies for malaria control and provided a powerful tool to investigate mosquito-parasite interactions. Our research on transmission blockade using transgenic mosquitoes has been focused on the midgut stage of the parasites, interfering with ookinete invasion or oocyst differentiation. The molecule that we have introduced into a transgenic mosquito is CEL-III isolated from Cucumaria echinata (Marine invertebrate). CEL-III has strong hemolytic activity as well as haemaggrutination activity as a lectin. CEL-III gene expression was driven by the carboxypeptidase promoter, midgut specific promoter. As expected, CEL-III expression was induced in the midgut at 6h after a blood meal. We fed control and transgenic mosquitoes on the same infected rat and measure the numbers of oocysts formed. The transmission blocking efficacy of the transgenic mosquitoes was 91%, as compared with control mosquito. Thus, expression of CEL-III in the mosquito midgut severely reduced vector competency by inhibiting Plasmodium development. Because CEL-III targets to red blood cells but not the parasites, we anticipate that this CEL-III transgenic mosquito will be able to impair parasite development of all Plasmodium species. Less