Cis-regulatory element discovery in a sequenced mosquito genome

 描述（由申请人提供）：媒介蚊子比任何其他生物都给人类带来更多的痛苦，每年杀死100多万人。蚊子基因组计划促进了蚊子生物学新方面的研究，包括我们对发育的功能遗传研究。尽管蚊子基因研究取得了重大进展，但对蚊子基因表达的调控仍然知之甚少。例如，很少有顺式调控元件（克雷斯），控制基因表达的DNA序列，已被确定在蚊子。由此产生的缺陷--蚊子遗传学基础知识的一个重大缺口--导致缺乏在特定时间操纵选定组织中基因表达的驱动程序。这些工具彻底改变了遗传模式生物的研究，将促进遗传研究，并有利于蚊子研究的所有途径。FAIRE-seq，与DNA测序配对的调节元件的甲醛辅助分离，正在成为全球CRE发现的强大的新的高通量工具。我们最近在登革热和黄热病媒介蚊子埃及伊蚊中进行了FAIRE-seq研究。这些努力的结果是在整个A.埃及人基因组这项研究将检验这些A.埃及伊蚊FAIRE DNA序列作为克雷斯的功能，包括在转基因昆虫中的高通量筛选，其将检查数百个这些元件在体内促进基因表达的能力。除了验证FAIRE数据集之外，筛选还被设计为选择具有高潜力的元件，以驱动多个载体昆虫物种中载体重要性组织中的基因表达。这项研究将产生一个基因驱动工具包 在蚊子研究方面，促进在其他媒介昆虫物种中使用FAIRE-seq，并鼓励研究蚊子基因调控网络。这项工作有可能改变媒介生物学家对待科学的框架。

 DESCRIPTION (provided by applicant): Vector mosquitoes inflict more human suffering than any other organism-and kill more than one million people each year. The mosquito genome projects have facilitated research in new facets of mosquito biology, including our functional genetic studies of development. Despite substantial progress in mosquito genetic research, regulation of mosquito gene expression is still poorly understood. For example, few cis-regulatory elements (CREs), DNA sequences that control gene expression, have been identified in mosquitoes. The resulting deficiency-a significant gap in the basic knowledge of mosquito genetics-has resulted in a lack of drivers to manipulate gene expression in selected tissues at specific times. Such tools, which have revolutionized research in genetic model organisms, would facilitate genetic studies and benefit all avenues of mosquito research. FAIRE-seq, formaldehyde-assisted isolation of regulatory elements paired with DNA sequencing, is emerging as a powerful new high-throughput tool for global CRE discovery. We recently performed a FAIRE-seq study in the dengue and yellow fever vector mosquito Aedes aegypti. These efforts resulted in identification of thousands of putative regulatory elements throughout the A. aegypti genome. The proposed investigation, which will test the hypothesis that these A. aegypti FAIRE DNA sequences function as CREs, includes a high throughput screen in transgenic insects that will examine the ability of hundreds of these elements to promote gene expression in vivo. In addition to validating the FAIRE data set, the screen has been designed to select for elements that have a high potential to drive gene expression in tissues of vector importance in multiple vector insect species. This research will generate a toolkit of gene drivers for mosquito research, promote use of FAIRE-seq in additional vector insect species, and encourage the study of mosquito gene regulatory networks. This work has the potential to change the framework in which vector biologists approach science.