The role of mosquito microRNAs in blood digestion.

亚历山大·S·莱赫尔 项目摘要/摘要 蚊媒疾病的破坏性是巨大的，约有100万人死亡，数百人死亡 每年有数以千计的人患重病。作为登革热、黄热病的主要传播媒介， 基孔肯雅病毒和最近的寨卡病毒，埃及伊蚊是一种极其重要的生物 调查。小的非编码microRNAs(MiRNA)负责mRNAs的转录后调控 并作为发展的基本调节者。我们在上一个资金周期中的努力揭示了 MiRNAs在控制雌性蚊子多种生理功能中的作用。此外，我们有 建立了一些基本的蚊子miRNA调查方法，从生物信息学和 MiRNA文库测序到遗传学方法，如CRISPR/Cas9和GAL4/UAS。这些研究已经 为进一步探索蚊虫miRNAs奠定了基础。然而，我们对miRNA角色的了解 在雌性蚊子这样一个必不可少的过程中，繁殖仍然有限。因此，目前赠款的目标是 建议阐明miRNAs参与管理血餐激活的调控网络 雌性蚊子的繁殖事件。总体假设是miRNAs是 这些监管网络。我们将在具体目标中检验以下假设：目标1.miRNAs是 参与血液消化所必需的酶的转录后调节。目标2.需要miRNAs 为了控制转录网络的关键因素，这是协调雌性蚊子所必需的 生殖周期。目的3.作为重要的调节分子，miRNAs受到严格的控制，协调 它们的活动与蚊子繁殖的需求有关。这项拟议研究的创新之处在于 MiRNAs在蚊子繁殖中的作用及其广泛应用的综合分析 方法，如CRISPR-CAS9和GAL4-UAS。目前拨款建议的意义在于 建立关于miRNAs与调控网络控制的关联的新范式 蚊子繁殖。

Alexander S. Raikhel PROJECT SUMMARY/ABSTRACT The damaging effect of mosquito-borne diseases is colossal, with about a million deaths and hundreds of thousands of people becoming seriously ill annually. Being the principal vector for dengue, yellow fever, chikungunya virus and recently Zika virus, the Aedes aegypti mosquito is an extremely important organism for investigation. Small non-coding microRNAs (miRNA) are responsible for posttranscriptional control of mRNAs and act as essential regulators of development. Our efforts during the previous funding cycle have revealed the role of miRNAs in controlling multiple physiological functions in female mosquitoes. Moreover, we have established a number of essential methodologies for mosquito miRNA investigations, from bioinformatics and miRNA library sequencing to genetics approaches such as CRISPR/Cas9 and GAL4/UAS. These studies have provided a foundation for further exploration of mosquito miRNAs. However, our knowledge about miRNA roles in such an essential process as female mosquito reproduction is still limited. Thus, the goal of the present grant proposal is to elucidate the involvement of miRNAs in regulatory networks governing blood-meal-activated reproductive events in female mosquitoes. The overall hypothesis is that miRNAs are critical components of these regulatory networks. We will examine the following hypotheses in Specific Aims: Aim 1. miRNAs are involved in posttranscriptional regulation of enzymes essential for blood digestion. Aim 2. miRNAs are required for the control of key factors of transcriptional networks that are necessary for orchestrating the female mosquito reproductive cycle. Aim 3. As important regulatory molecules, miRNAs are under strict control, coordinating their activity with demands of mosquito reproduction. The innovation of this proposed research rests on a comprehensive analysis of the role of miRNAs in mosquito reproduction and application of wide-ranging methodologies, such as CRISPR-Cas9 and Gal4-UAS. The significance of the present grant proposal is in establishing a novel paradigm concerning the association of miRNAs with regulatory networks controlling mosquito reproduction.