Genetic tools for manipulation of mosquitoes

操纵蚊子的遗传工具

基本信息

批准号：
8515315
负责人：
KRISTEN A BERNARD
金额：
$ 17.26万
依托单位：
UNIVERSITY OF WISCONSIN-MADISON
依托单位国家：
美国
项目类别：
财政年份：
2012
资助国家：
美国
起止时间：
2012-08-01 至 2014-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8515315
关键词：
Adult Aedes Anopheles Genus Anopheles gambiae Area Behavior Biological Biological Assay Biology Culicidae DNA DNA Transposable Elements DNA delivery Dissection Ecology Elements Embryo Evolution Female Future Gene Expression Gene Expression Profiling Gene Transfer Techniques Generations Genetic Genetic Engineering Genetic Recombination Genetic Transformation Human Human Bites Injection of therapeutic agent Insecta Insecticide Resistance Larva Lethal Genes Malaria Measures Mediating Methods Midgut Mosquito Control Olfactory Pathways Oocytes Ovary Plasmids Procedures Process Public Health Reporter Reporter Genes Reporting Resources Staging System Testing Time Transgenic Organisms Vector-transmitted infectious disease base burden of illness disorder control egg gene function genome sequencing in vivo molecular vector novel pathogen plasmid DNA promoter research study success tool uptake vector vector mosquito

项目摘要

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.

描述（由申请人提供）：蚊子传播的病原体会引起巨大的人类痛苦。媒介蚊子的基因组序列是理解和开发媒介特异性过程以减轻疾病负担的重要资源。但是，对这些过程的研究的主要局限性是可用的基因表达在体内表达的可用遗传工具不足。我们的初步研究表明，基于女性蚊子生物学的关键元素，将DNA传递到胚胎的新方法的可行性。我们的研究记录了在将质粒递送到上演的女性之后，质粒DNA和报告基因活性从卵和成年阶段的持续存在。现在，我们建议以这种背景为基础，并测试“垂直输送方法”的潜在应用。在AIM 1中，我们将优化该方法，并确定报告基因的瞬时表达是否可用于鉴定卵巢和中肠的成年女性特异性启动子。在AIM 2中，我们将确定是否可以使用转座元件构建体的垂直输送来有效地生成AE的转基因线。埃及和。冈比亚。这些目标的成功完成将提供新的方法，这些方法将极大地促进对蚊子生物学的未来分析。