Genetic tools for manipulation of mosquitoes

描述（由申请人提供）：蚊媒病原体给人类带来巨大痛苦。病媒蚊子的基因组序列是了解和利用病媒特异性过程以减轻疾病负担的重要资源。然而，研究这些过程的一个主要限制是缺乏可用的遗传工具来操纵基因在体内的表达。我们的初步研究证明了一种基于雌蚊生物学关键要素的胚胎DNA传递新方法的可行性。我们的研究记录了埃及伊蚊和冈比亚按蚊的质粒DNA和报告基因活性从卵到成虫阶段的持久性，并将质粒交付给分期的雌性。我们现在建议在此背景下，测试“垂直交付方法”的潜在应用。在Aim 1中，我们将优化方法，并确定报告基因的瞬时表达是否可以用于鉴定卵巢和中肠的成年女性特异性启动子。在目标2中，我们将确定垂直递送转座元件构建体是否可用于有效地产生Ae转基因系。埃及伊蚊和安。冈比亚按蚊。这些目标的成功完成将为今后蚊子生物学的分析提供新的方法。

DESCRIPTION (provided by applicant): Mosquito-borne pathogens cause enormous human suffering. The genome sequences of vector mosquitoes are important resources for understanding and exploitation of vector-specific processes to reduce the burden of disease. However, a major limitation to studies of these processes is the inadequacy of available genetic tools for manipulating gene expression in vivo. Our preliminary studies demonstrate the feasibility of a new method for DNA delivery to embryos based on key elements of female mosquito biology. Our studies document the persistence of plasmid DNA and reporter gene activity from the egg to the adult stage in both Aedes aegypti and Anopheles gambiae, following delivery of the plasmids to the staged females. We now propose to build on this background and test potential applications of the "vertical delivery method". In Aim 1, we will optimize the method and determine whether transient expression of reporter genes can be used to identify adult female-specific promoters for the ovary and midgut. In Aim 2, we will determine whether vertical delivery of transposable element constructs can be used to efficiently generate transgenic lines of Ae. aegypti and An. gambiae. Successful completion of these aims will provide new methods that will greatly facilitate future analyses of mosquito biology.