Zika and dengue virus agonists for transmission-blocking

用于阻断传播的寨卡病毒和登革热病毒激动剂

• 批准号: 9469995
• 负责人: George Dimopoulos
• 金额: $ 20.44万
• 依托单位: JOHNS HOPKINS UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-12-15 至 2019-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9469995
• 关键词: Aedes; Agonist; Antibodies; Arboviruses; Biological Assay; Biology; Blood; CRISPR/Cas technology; Computer Retrieval of Information on Scientific Projects Database; Culicidae; Data; Dengue; Dengue Infection; Dengue Virus; Development; Fertility; Future; Gene Deletion; Gene Silencing; Gene Transfer Techniques; Genes; Goals; Impairment; Infection; Institution; Integration Host Factors; Knock-out; Knowledge; Literature; Longevity; Mediating; Methodology; Midgut; Modeling; Nutritional; Pharmaceutical Preparations; Principal Investigator; RNA Interference; Research; Research Infrastructure; Resistance; Role; Saliva; System; Technology; Testing; Time; TimeLine; Tissues; Transgenic Organisms; Virus; Virus Diseases; Work; ZIKV infection; Zika Virus; base; disorder control; experience; feeding; fitness; functional genomics; novel; permissiveness; screening; small molecule; transmission process; vector; vector mosquito; viral transmission

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.

首席研究员的长期研究目标是阐明蚊子载体与寨卡病毒（ZIKV）和登革热病毒（DENV）之间的相互作用，以确定适合制定疾病控制策略的传播阻断靶点。在通过媒介传播的过程中，病毒依赖于许多蚊子衍生的激动剂（宿主因子），与蚊子的中肠和其他组织进行密切的相互作用。ZIKV和DENV可以通过基因编辑删除激动剂，通过RNA干扰耗尽，或通过抗体阻断/干扰/抑制，在载体中受损。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手段靶向。最近在伊蚊中开展的基于CRISPR/ cas9的基因编辑为通过基因缺失研究激动剂功能提供了新的和有希望的机会，并可用于开发新的ZIKV和DENV控制策略。在这里，我们将重点关注10种病毒激动剂，以评估它们的疾病控制潜力，并进一步了解它们在寨卡病毒和登革热病毒感染中的作用。在目标1中，我们将验证6种假定的DENV激动剂对寨卡病毒的阻断潜力。利用rnai介导的基因沉默。在Aim 2中，我们将为选定的ZIKV和DENV激动剂培养CRISPR/ cas9介导的条件血粉诱导的中肠特异性基因KO蚊子。在目标3中，我们将评估这些KO Ae。埃及伊蚊对病毒感染的抗性及其对蚊子寿命和繁殖力的适应性影响。