Guy1-conferred female lethality: mechanism and application for mosquito control

 描述（由申请人提供）：按蚊是疟疾的主要媒介，疟疾是人类最致命和代价最高的疾病之一。随着抗药性和杀虫剂抗药性的蔓延，疟疾控制措施受到越来越大的威胁。最近开发的基因组资源和遗传方法提供了前所未有的机会，以提高我们对蚊子生物学的理解，并制定新的战略来对抗疟疾和其他蚊媒疾病。尽管Y染色体在雄性决定和雄性生物学的其他方面很重要，但它在按蚊中的作用还没有得到研究。由于吸血和因此传播疾病病原体是雌性蚊子的唯一领域，因此在实施减少蚊子种群或用不易传播疾病的种群取代有能力的病媒种群的遗传方法时，只释放雄性蚊子至关重要。克服技术瓶颈，我们最近在An中发现了四个Y基因。史蒂芬西其中，Guy 1是男性决定性M因子的最佳候选者。产生了多个表达Guy 1的转基因系，其在多代中产生了6520个转基因雄性和0个转基因雌性。我们的初步数据还表明，在Guy 1转基因An. Stephensi可能由剂量补偿的错误调节引起。重要的是，三个已知的性别决定主开关，性别致死，Fem/Masc，xo-lethal 1，也直接或间接调节剂量补偿在果蝇，家蚕，和秀丽隐杆线虫，分别。性致死等位基因的功能丧失和Masc靶基因的敲低导致雌性胚胎死亡。melanogaster和B.森，分别。在这里，我们将测试的假设，一个。stephensi Guy 1是性别决定或剂量补偿或两者的主调节器。我们将追求以下具体目标：1）确定Guy 1是否控制剂量补偿和/或性别决定; 2）描述Guy 1功能的分子机制; 3）利用Guy 1转基因作为按蚊性别分离和种群减少的手段。

 DESCRIPTION (provided by applicant): Anopheles mosquitoes are the primary vectors of malaria, one of humankind's most deadly and costly diseases. Malaria control measures are under increasing threat as drug- and insecticide- resistance spreads. Recently developed genomic resources and genetic methods provide unprecedented opportunities to improve our understanding of mosquito biology and develop new strategies to combat malaria and other mosquito-borne diseases. Y chromosome remains unexplored in Anopheles mosquitoes despite its importance in male determination and other aspects of male biology. Since blood feeding and, consequently, the transmission of disease pathogens is the sole province to female mosquitoes, it is critical to release only males when implementing genetic approaches for reducing mosquito populations or for replacing competent vector populations with populations that are refractory to disease transmission. Overcoming technical bottlenecks, we have recently uncovered four Y genes in An. stephensi. Among these, Guy1 is the best candidate for the male-determining M factor. Multiple Guy1-expressing transgenic lines were generated, which produced 6520 transgenic males and 0 transgenic females over multiple generations. Our preliminary data also suggest that the observed 100% female lethality in the Guy1 transgenic An. stephensi may result from mis-regulation of dosage compensation. Importantly, three known master switches of sex-determination, sex-lethal, Fem/Masc, xo-lethal 1, also directly or indirectly regulate dosage compensation in Drosophila melanogaster, Bombyx mori, and Caenorhabditis elegans, respectively. Loss of function sex- lethal alleles and knockdown of the target of Masc cause female embryonic lethality in D. melanogaster and B. mori, respectively. Here, we will test the hypothesis that the An. stephensi Guy1 is a master regulator of either sex-determination or dosage compensation or both. We will pursue the following specific aims: 1) To determine whether Guy1 controls dosage compensation and/or sex-determination; 2) To delineate the molecular mechanism underlying Guy1 function; and 3) To exploit Guy1 transgenics as a means for sex separation and population reduction in Anopheles mosquitoes.