Sex separation through targeting of conserved female-specific lethal genes in disease vector mosquitoes

项目摘要 雌蚊在形态、生理和行为特征上与雄蚊不同， 致病病原体的传播。尽管大多数调控两性异形的基因 蚊子的发育尚未确定，这些基因可能代表了新的靶点， 灭蚊。雌性特异性发育基因的表征也可以阐明遗传学 雄性蚊子的生产和分类战略，这是阻碍全球执行 几种新出现的基于人口的蚊子控制战略依赖于大规模释放雄性蚊子。 导致性别特异性致死效应的基因座簇被认为位于性别决定基因M/m内。 基因座区域的登革热载体蚊子埃及伊蚊，但这些基因的身份是不是 以前知道的。最近的一项高通量RNAi介导的筛选揭示了雌性特异性幼虫致死性 A.埃及人性别决定基因座区域，包括几个在其他基因中具有已知直系同源物的基因。 病媒蚊子的种类。针对这些基因的酵母干扰RNA杀幼虫剂杀死雌性A。 埃及伊蚊幼虫发育期间，但不影响雄性蚊子。拟议的研究计划将 测试假设，这是支持的初步数据，即这些基因的直系同源物也发挥作用， 白纹伊蚊（多种虫媒病毒的载体）、冈比亚按蚊的雌性特异性幼虫致死位点 （疟疾媒介）和致倦库蚊（西尼罗河和淋巴丝虫病媒介）。的具体目标 提出的研究包括i）RNAi介导的这些中的预期女性特异性致死基因的筛选， 疾病媒介蚊子，ii）构建和向下选择雌性特异性酵母杀幼虫剂以靶向 基因在每一个物种，和iii）开发的协议，酵母幼虫为基础的大量饲养适合，交配- 三种蚊子都有竞争力预计这些研究将产生新的 雌性特异性杀幼虫控制策略，提供了对蚊子性染色体进化的深入了解 和性别规范机制，并阐明方法，大规模饲养的男性，这将有助于 在多种人类病媒蚊子中，正在出现基于人口的控制战略。

Project Summary Female mosquitoes differ from males in morphological, physiological, and behavioral traits that promote the spread of disease-causing pathogens. Although the majority of genes that regulate sexually dimorphic development in mosquitoes have not yet been identified, these genes may represent novel targets for mosquito control. Characterization of female-specific developmental genes could also elucidate genetic strategies for production and sorting of male mosquitoes, an obstacle that hinders global implementation of several emerging population-based mosquito control strategies that are dependent on mass releases of males. Clusters of loci that cause sex-specific lethal effects are believed to reside within the sex-determining M/m locus region of the dengue vector mosquito Aedes aegypti, but the identities of these genes were not previously known. A recent high-throughput RNAi-mediated screen uncovered female-specific larval lethal genes in the A. aegypti sex-determining locus region, including several genes with known orthologs in other species of disease vector mosquitoes. Yeast interfering RNA larvicides that target these genes kill female A. aegypti larvae during development, but do not impact male mosquitoes. The proposed research program will test the hypothesis, which is supported by preliminary data, that orthologs of these genes also function as female-specific larval lethal loci in Aedes albopictus (vector of multiple arboviruses), Anopheles gambiae (malaria vector), and Culex quinquefasciatus (West Nile and lymphatic filariasis vector). Specific aims of the proposed investigation include i) RNAi-mediated screening of prospective female-specific lethal genes in these disease vector mosquitoes, ii) construction and down-selection of female-specific yeast larvicides to target the genes in each species, and iii) development of protocols for yeast larvicide-based mass rearing of fit, mating- competitive male mosquitoes of all three species. It is anticipated that these studies will generate novel female-specific larvicidal control strategies, provide insight into the evolution of mosquito sex chromosomes and sex-specification mechanisms, and elucidate methodology for mass rearing of males that will facilitate emerging population-based control strategies in multiple species of human disease vector mosquitoes.