Role of cell tropism for Zika virus transmission and pathogenesis

该提案是针对R21，细胞嗜性对寨卡病毒发病机制和载体能力的作用。的 该项目是格雷戈里·埃贝尔博士（科罗拉多州立大学）和亚伦·布劳特博士（ 疾病控制和预防）。这项研究的总体目标是了解不同的细胞 导致寨卡病毒的性传播和蚊子传播。我们对这个项目的总体方法包括使用一个 新开发的寨卡病毒PRABC59株的cDNA克隆（来自波多黎各患者，2015年）， 已经被工程化以表达miRNA靶序列。这些序列限制了病毒在生物体中的复制- 组织和细胞特异性的方式，由于结合的同源miRNA的病毒RNA和随后的 通过RNAi途径降解vRNA。 目的1着重于开发合适的构建体，并使用细胞系和原代细胞评估功效。 cultures.目标2将该项目转移到使用Ag129小鼠的体内测试阶段，这些小鼠可在CDC获得 作为繁殖群体，和埃及伊蚊，这是在科罗拉多州立大学提供的几个新建立的 殖民地 拟议的研究利用了广泛的初步数据和发展中的合作， 在分子病毒学、虫媒病毒发病机理和载体生物学方面的专业知识。

This proposal is for an R21, Role of cell tropism for pathogenesis and vector competence of Zika virus. The project is a collaboration between Drs. Gregory Ebel (Colorado State University) and Aaron Brault (Centers for Disease Control and Prevention). The overall goal of this research is to understand how different cells contribute to sexual and mosquito transmission of Zika. Our overall approach to this project involves using a newly developed cDNA clone of the PRABC59 strain of Zika virus (from a Puerto Rican patient, 2015) that has been engineered to express miRNA target sequences. These sequences restrict virus replication in organism- tissue- and cell-specific manners due to binding of the cognate miRNA to viral RNA and subsequent degradation of the vRNA by the RNAi pathway. Aim 1 focuses on developing appropriate constructs and evaluating efficacy using cell lines and primary cultures. Aim 2 moves the project into an in vivo testing phase using Ag129 mice, which are available at CDC as a breeding colony, and Aedes aegypti mosquitoes, which are available at CSU as several newly established colonies. The proposed research leverages extensive preliminary data and a developing collaboration that brings together expertise in molecular virology, arboviral pathogenesis and vector biology.