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

通过改进的基于 CRISPR 的全基因组敲除筛选来识别 ZIKV 宿主因子

• 批准号: 9265173
• 负责人: Haoquan Wu
• 金额: $ 19.13万
• 依托单位: TEXAS TECH UNIVERSITY HEALTH SCIS CENTER
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-09-05 至 2018-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9265173
• 关键词: Attention; Biological Process; Cell Death; Cells; Clinical; Data Quality; Disease Outbreaks; Endoplasmic Reticulum Degradation Pathway; Ensure; Essential Genes; Family; Family member; Flavivirus; Genes; Genome engineering; Health; Human; Individual; Integration Host Factors; Investigation; Knock-out; Knowledge; Libraries; Life Cycle Stages; Maps; Mediating; Methods; Paper; Pathway interactions; Performance; Phenotype; Process; Public Health; RNA Interference; Regulation; Reporting; Research; Resistance; Sensitivity and Specificity; Series; System; Technology; Viral; Virus; Virus Replication; West Nile virus; Zika Virus; base; design; endonuclease; genome-wide; genome-wide analysis; improved; killings; knockout gene; pathogen; screening; therapeutic target; virus host interaction

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.

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。可是 对这种病毒知之甚少，包括它如何复制和杀死宿主细胞。无偏 全基因组功能筛选促进病毒复制并随后产生的宿主因子 细胞死亡将为更深入地探索宿主-病毒相互作用提供一条途径，因为病毒依赖于 在生命周期的每一步都有宿主因素的影响。在这里，我们建议系统地识别ZIKV 利用基于CRISPR的全基因组敲除筛选方法， Cas9是我们独立开发的。在初步研究中，我们已经确定了宿主 具有高灵敏度和特异性的因素，更重要的是，西方的强表型 尼罗河病毒（WNV），其是ZIKV的密切黄病毒家族成员。此外，我们还力求 改进我们的筛选策略，使具有较弱表型的宿主基因也能被 鉴定因此，我们希望识别ZIKV宿主因素，以构建全面的人类 宿主因子-病毒相互作用图谱，并鉴定ZIKV的潜在治疗靶标。