Identify ZIKV host factors with an improved CRISPR-based genome-wide knockout screen

摘要 持续的寨卡病毒(ZIKV)疫情在全球范围内构成了严重的健康挑战。然而，非常 人们对这种病毒知之甚少，包括它如何复制和杀死宿主细胞。不偏不倚的 对促进病毒复制和随后的宿主因子的全基因组功能筛选 细胞死亡将为更深入地探索宿主与病毒的相互作用提供一条途径，因为病毒依赖于 它们生命周期中每一步的寄主因素。在这里，我们建议系统地识别ZIKV 基于CRISPR的全基因组基因敲除筛选方法在人类细胞中的宿主因子 CAS9是我们自主开发的。在初步研究中，我们已经确定了宿主 具有高敏感度和特异度的因子，更重要的是，西方的强表型 尼罗河病毒(Nile Virus，WNV)是ZIKV的一个亲缘关系密切的黄病毒家族成员。此外，我们还寻求 改进我们的筛选策略，以便表型较弱的宿主基因也将 已确认身份。因此，我们期望找出ZIKV的宿主因素，构建一个全面的人类 宿主因子-病毒相互作用图，并确定ZIKV的潜在治疗靶点。

Abstract Ongoing Zika virus (ZIKV) outbreaks pose a serious health challenge worldwide. However, very little is known about this virus, including how it replicates and kills host cells. An unbiased genome-wide functional screen for the host factors that facilitate viral replication and ensuing cell death would provide a path to explore host-virus interaction more deeply since viruses rely on host factors for every step of their life cycle. Here we propose to systematically identify ZIKV host factors in human cells with a genome-wide knockout screening method based on CRISPR- Cas9 that we have developed independently. In preliminary studies, we have identified host factors with high sensitivity and specificity and more importantly, strong phenotypes for West Nile virus (WNV), which is a close flavivirus family member of ZIKV. In addition, we also seek to improve our screening strategy so that host genes with weaker phenotypes will also be identified. Thus, we expect to identify ZIKV host factors to construct a comprehensive human host factor–virus interaction map, and to identify potential therapeutic targets for ZIKV.