Identification and validation of Zika virus receptor(s) in the midgut lumen of Aedes aegypti

埃及伊蚊中肠腔中寨卡病毒受体的鉴定和验证

• 批准号: 10379934
• 负责人: Sujit K Pujhari
• 金额: $ 18.94万
• 依托单位: UNIVERSITY OF SOUTH CAROLINA AT COLUMBIA
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-04-01 至 2024-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10379934
• 关键词: Ablation; Aedes; Africa; Arbovirus Infections; Arboviruses; Asia; Autoimmune Diseases; Binding; Biochemical; Biological Assay; Biology; Blood; Brain; Breathing; CRISPR/Cas technology; Cell Surface Receptors; Centers for Disease Control and Prevention (U.S.); Cessation of life; Country; Culicidae; DNA Sequence; Data; Development; Disease; Disease Outbreaks; Epidemic; Epithelial; Epithelial Cells; Flavivirus; Future; Generations; Genes; Genetic; Goals; Grant; Guide RNA; Guillain Barré Syndrome; Human; Impairment; Infant; Infection; Insecta; Intervention; Investigation; Island; Mass Spectrum Analysis; Measures; Mediating; Membrane; Membrane Proteins; Methods; Micronesia; Midgut; Molecular; Molecular Virology; Mosquito Control; Muscle Weakness; Oral; Organism; Paralysed; Placenta; Procedures; Process; Protein Engineering; Proteins; Proteome; Protocols documentation; Recombinants; Research; Research Personnel; Role; Surface; Symptoms; Talents; Testing; Therapeutic; Training; Transgenic Organisms; Uganda; Update; Vaccines; Validation; Vector-transmitted infectious disease; Virus; Virus Receptors; Wolbachia; Work; ZIKA; ZIKV infection; Zika Virus; arthropod-borne; design; disorder control; endonuclease; env Gene Products; experience; experimental study; fetal; forest; improved; in vivo; insight; mosquito-borne; mutant; novel; pathogen; protein complex; receptor; transmission process; transmission-blocking vaccine; vaccine candidate; vaccine development; vector; vector competence; vector mosquito; viral transmission

SUMMARY The central goal of this project is to identify Zika virus (ZIKV) receptor(s), generation ZIKV transmission incompetent transgenic mosquitoes that lack the receptor-encoding gene and identification of novel transmission blocking vaccine candidate(s). Transmission of arthropod-borne viruses (arboviruses) causes widespread and debilitating diseases across the globe. In the absence of effective licensed vaccines or therapeutic treatments of arboviruses, including ZIKV, control at the vector level is the most effective method to ameliorate the burden of these viruses. Before these viruses can be transmitted to humans, they must first infect the mosquito following a blood meal. Thus, investigations to understand ZIKV-mosquito interactions are of critical importance. In this proposal, we will identify specific molecules in the Aedes aegypti mosquito midgut that act as receptors and facilitators for ZIKV binding and establishment of infection. Identified candidate receptors will be validated in vivo by genetic ablation using CRISPR/Cas9 gene editing and subsequent oral challenge with an infectious ZIKV blood meal. Data generated in this study will assist in 1) the development of effective disease control measures, e.g. via the generation of transgenic mosquitoes that lack the receptor-encoding gene and would be incompetent for ZIKV transmission; and 2) identification of ZIKV receptor molecules in the mosquito midgut will provide new insights into the basic biology and molecular mechanisms underpinning the vector competency and mosquito- arboviral pathogen interactions. The proposed experiments are also significant because they provide the groundwork for future experiments that will determine whether the same receptor(s) is used by other arboviruses and other mosquito species.

总结 该项目的中心目标是确定寨卡病毒（ZIKV）受体，产生ZIKV传播 缺乏受体编码基因的无能转基因蚊子和新传播的鉴定 阻断候选疫苗。节肢动物传播的病毒（虫媒病毒）的传播引起广泛的， 使人衰弱的疾病在地球仪上传播。在缺乏有效的许可疫苗或治疗方法的情况下， 虫媒病毒，包括ZIKV，在载体水平上控制是改善虫媒病毒负担的最有效方法。 这些病毒。在这些病毒可以传播给人类之前，它们必须首先感染蚊子， 血餐因此，了解ZIKV与蚊子相互作用的研究至关重要。在这 根据这项提议，我们将确定埃及伊蚊中肠中作为受体的特定分子， ZIKV结合和建立感染的促进剂。将在体内验证已鉴定的候选受体 通过使用CRISPR/Cas9基因编辑的基因消融和随后的感染性ZIKV的口服攻击， 血餐本研究产生的数据将有助于1）制定有效的疾病控制措施， 例如通过产生缺乏受体编码基因的转基因蚊子， 2）蚊子中肠中ZIKV受体分子的鉴定将为ZIKV的传播提供新的途径。 深入了解支持媒介能力和蚊子的基本生物学和分子机制， 虫媒病毒病原体相互作用。所提出的实验也很重要，因为它们提供了 为将来的实验奠定基础，以确定其他虫媒病毒是否使用相同的受体 和其他蚊子物种。