Zika and dengue virus agonists for transmission-blocking

首席研究员研究的长期目标是阐明蚊媒与寨卡病毒(ZIKV)和登革病毒(DENV)之间的相互作用，确定适合制定疾病控制战略的传播阻断目标。在通过媒介的旅程中，病毒依赖于许多蚊子衍生的激动剂(宿主因素)与蚊子中肠和其他组织进行亲密的相互作用。载体中的ZIKV和DENV可通过基因编辑的激动剂缺失、RNA干扰的耗尽或抗体的阻断/干扰/抑制而受损。因此，研究病毒激动剂用于疾病控制的一个明显优势是，它们可以通过多种方式被靶向。最近发展起来的基于CRISPR/Cas9的伊蚊基因编辑为通过基因缺失来研究激动剂功能提供了新的机会，并可用于开发新的ZIKV和DENV控制策略。在这里，我们将重点介绍10种病毒激动剂，以评估它们在疾病控制方面的潜力，并进一步了解它们与寨卡病毒和登革热病毒感染的关系。在目标1中，我们将验证6种推定的DENV激动剂对Ae中ZIKV的阻断潜力。埃及伊蚊利用RNAi介导的基因沉默。在目标2中，我们将为选定的ZIKV和DENV激动剂开发CRISPR/Cas9介导的条件性血粉诱导的中肠特异基因KO蚊子。在目标3中，我们将评估这些KO Ae。埃及伊蚊品系对病毒感染的抵抗力及其对蚊子寿命和繁殖力的适合性影响。

The long-term goal of the principal investigator’s research is to elucidate the interactions between the mosquito vector and Zika virus (ZIKV) and Dengue virus (DENV), to identify transmission-blocking targets suitable for the development of disease control strategies. During their journey through the vector, the viruses engage in intimate interactions with the mosquito midgut and other tissues, relying on numerous mosquito-derived agonists (host factors). ZIKV and DENV can be impaired in the vector by agonist deletion through gene editing, depletion through RNA interference, or blocking / interference / inhibition through antibodies. Hence, an obvious advantage of studying virus agonists for disease control is that they can be targeted through multiple means. The recently developed CRISPR/CAS9-based gene editing in Aedes mosquitoes provide new and promising opportunities for the study of agonist function through gene deletion, and could be used to develop novel ZIKV and DENV control strategies. Here we will focus on 10 virus agonists to assess their potential for disease control and further our knowledge of their involvement in ZIKV and DENV infection. In Aim 1, we will validate 6 putative DENV agonists for ZIKV blocking potential in Ae. aegypti using RNAi-mediated gene silencing. In Aim 2, we will develop CRISPR/CAS9-mediated conditional bloodmeal-inducible midgut-specific gene KO mosquitoes for selected ZIKV and DENV agonists. In Aim 3, we will assess these KO Ae. aegypti lines for resistance to virus infection and their fitness impact in terms of mosquito longevity and fecundity.