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通过基因组尺度CRISPR选进行Flaviviridae宿主因子的遗传剖析

Caleb D Marceau, Andreas S Puschnik, Karim Majzoub

|July 8, 2016

概括

这项研究揭示了登革热病毒 (DENV) 和型肝炎病毒 (HCV) 复制所必需的不同宿主因素. 针对这些宿主依赖,如DENV的寡糖转移酶复合体,提供了新的抗病毒策略.

科学领域:

病毒学
分子生物学
遗传学

背景情况:

包括登革热病毒 (DENV) 和型肝炎病毒 (HCV) 在内的Flaviviridae病毒造成了严重的全球健康负担,治疗选择有限.
了解宿主-病原体相互作用对于开发有效的抗病毒疗法来对抗这些新出现和持续的病毒威胁至关重要.

研究的目的:

采用聚合CRISPR遗传查方法全面识别DENV和HCV复制所必需的宿主因素.
阐明已识别的宿主因子在病毒复制机制中的特定作用.
探索DENV和HCV之间宿主依赖的潜在分歧和趋同.

主要方法:

使用集成的CRISPR基因查策略系统地破坏宿主基因并评估它们对DENV和HCV复制的影响.
进行了机理学研究,以确定病毒生命周期阶段依赖于已识别的宿主因素.
研究病毒蛋白与宿主复合体之间的相互作用.

主要成果:

确定了与内质网膜 (ER) 相关的蛋白质复合体,包括寡糖转移酶 (OST) 复合体,对于DENV复制,特别是病毒RNA复制至关重要.
证明OST复合体缺陷消除了DENV复制,病毒蛋白与OST相互作用.
发现了HCV的独特宿主因素,包括病毒受体,RNA结合蛋白,代谢酶以及与黄氨酸二核酸 (FAD) 水平的意外联系.

结论:

对DENV和HCV的宿主因子要求存在显著差异,突出了病毒特异性的脆弱性.
OST复合体是DENV和其他蚊子传播的黄病毒的关键宿主因子,是潜在的抗病毒点.
鉴定到的宿主依赖因素为开发抗病毒疗法提供了新的点.

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