Cis-regulatory element discovery in a sequenced mosquito genome

 描述(申请人提供)：媒介蚊子给人类造成的痛苦比任何其他生物体都多--每年导致100多万人死亡。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学研究。尽管蚊子基因研究取得了实质性进展，但对蚊子基因表达的调控仍然知之甚少。例如，在蚊子中几乎没有发现顺式调控元件(CRE)，也就是控制基因表达的DNA序列。由此导致的缺陷--蚊子遗传学基础知识的巨大差距--导致缺乏在特定时间操纵选定组织中基因表达的驱动因素。这些工具彻底改变了遗传模型生物的研究，将促进遗传研究，并使蚊子研究的所有途径受益。FIRE-SEQ，甲醛辅助的调节元件分离与DNA测序相结合，正在成为一种强大的新的高通量工具，用于全球Cre发现。我们最近在登革热和黄热病媒介蚊子埃及伊蚊中进行了一项FIRE-SEQ研究。这些努力最终确定了埃及伊蚊基因组中数千个可能的调控元件。这项拟议中的研究将检验埃及斑潜蝇DNA序列作为Cres功能的假设，包括在转基因昆虫中进行高通量筛选，以检查数百种这些元素在体内促进基因表达的能力。除了验证FIRE数据集之外，该筛查还被设计用于选择具有高潜力驱动多种媒介昆虫物种中具有媒介重要性的组织中的基因表达的元件。这项研究将产生一个基因驱动程序工具包 在蚊子研究方面，推广在更多媒介昆虫物种中使用FIRE-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接近科学的框架。

 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.