Generation of transmission-compromised mosquitoes

昆虫传播的疾病导致的死亡人数超过了所有其他疾病的总和。现有的战略限制 这类疾病的传播是有帮助的，但很明显，彻底根除一些最致命的 疾病--尤其是疟疾--将需要更多的方法。该项目的目标是测试 通过在昆虫媒介水平进行干预来阻止虫媒疾病传播的策略，预防 通过导致成年雌性及时死亡来传播致病寄生虫和病毒 蚊子唾液腺(SG)。当前对试图操纵SG-a网关的研究的限制 传播疟疾和其他昆虫传播疾病的器官--关于SG几乎一无所知 果蝇以外的任何昆虫的发育、功能或动态平衡。参与指定的分子， 建造或专门化蚊子SGS在很大程度上是未知的。拟议的研究首先了解 冈比亚按蚊转录因子(Tf)基因RNAi敲除的后果 果蝇同源基因在果蝇SG的发育、功能和生存中起着关键作用。我们问特工队是否 基因敲除会影响SG的存活或形态，或感染疟疾的子孢子侵袭 SGS并通过唾液传播。在第二个目标中，我们在 冈比亚按蚊的sage基因，它编码一种SG特异性的转铁蛋白，它的果蝇同源基因需要 SG的存活和SG特有的Cargo基因的表达。我们问，完全丧失SAGE对生存能力有何影响， 繁殖力和SG形态。我们询问SAGE的丢失是否影响子孢子SG的入侵和子孢子在 唾液。我们还确定了血餐后特定于SG的SAGE缺失是否会导致类似的结果。 最后，为了了解我们以SG为中心的方法在总体上限制疾病传播的效果如何，我们询问 埃及伊蚊SAGE同源基因的RNAi敲除影响SG及其传播登革热的能力 病毒。

Insect-borne diseases kill more humans than all other diseases combined. Existing strategies for limiting the spread of such diseases are helping, but it is clear that complete eradication of some of the most deadly diseases – especially malaria – is going to require additional approaches. The goal of this project is to test strategies to block insect-borne disease transmission by intervening at the level of the insect vector, preventing transmission of disease-causing parasites and viruses by causing the timely death of the adult female mosquito salivary gland (SG). A current limitation for studies attempting to manipulate the SG – a gateway organ for transmitting malaria and other insect borne diseases - is that almost nothing is known about SG development, function or homeostasis in any insect outside Drosophila. The molecules involved in specifying, building or specializing mosquito SGs are largely unknown. The proposed study begins with learning the consequences of RNAi knock-down of three Anopheles gambiae transcription factor (TF) genes whose Drosophila orthologues have key roles in Drosophila SG development, function and survival. We ask if TF gene knockdown affects SG survival or morphology, or the ability of infective malaria sporozoites to invade the SGs and to be transmitted in the saliva. In the second aim, we create null and tissue-specific mutations in the Anopheles gambiae sage gene, which encodes a SG-specific TF whose Drosophila orthologue is required for SG survival and expression of SG-specific cargo genes. We ask how complete loss of sage affects viability, fecundity and SG morphology. We ask if loss of sage affects sporozoite SG invasion and sporozoite counts in saliva. We also determine if SG-specific loss of sage following a blood meal leads to similar outcomes. Finally, to learn how well our SG-centric approach to limiting disease transmission works in general, we ask if RNAi knockdown of the sage orthologue in Aedes aegypti affects the SG and its ability to transmit Dengue virus.