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基于复制体的全基因组CRISPR淘汰查用于识别参与病毒复制的宿主因素

Karen W Cheng¹, Madhura Bhave¹, Andrew L Markhard^1,2

¹Chan Zuckerberg Biohub, San Francisco, CA, USA.

Nature communications

|December 10, 2025

概括

这项研究引入了一种新的病毒复制CRISPR淘汰查方法,以找到病毒复制的必要宿主因素. 这种方法成功地识别了登革热病毒,奇孔古尼亚病毒和埃博拉病毒的已知和新宿主基因.

科学领域:

病毒学病毒学
遗传学是一个遗传学.
分子生物学分子生物学

背景情况:

识别对病毒复制至关重要的宿主因素对于开发抗病毒疗法至关重要.
传统的活病毒选通常会因为安全问题或检测局限性而忽略关键宿主因素.
克里斯普尔淘汰 (KO) 查为功能基因组学提供了强大的工具,但其在病毒研究中的应用需要特定的适应.

研究的目的:

开发和验证一种基于病毒复制的CRISPR KO查方法,用于识别病毒复制必不可少的宿主因素.
使用登革热病毒2型 (DENV-2) 复制系统对查方法进行基准测试.
将这种查方法的应用扩展到其他RNA病毒,包括 Chikungunya病毒 (CHIKV) 和埃博拉病毒 (EBOV).

主要方法:

稳定的光登革热病毒2型 (DENV-2) 复制细胞系的发展.
将CRISPR KO选应用于DENV-2复制体系统,以识别重要的宿主基因.
将基于复制体的CRISPR KO查扩展到奇孔古尼亚病毒 (CHIKV) 和埃博拉病毒 (EBOV) 系统.
对已识别的宿主因子及其在病毒复制中的作用进行分析.

主要成果:

通过DENV-2复制体查,成功地确定了已知的宿主因素,包括内细胞网膜复合体和寡糖转移酶复合体组件.
屏幕还揭示了以前在CRISPR KO屏幕中与DENV-2复制无关的新型宿主基因.
使用基于复制体的查方法确定了对CHIKV和EBOV复制必不可少的不同组宿主基因.
该方法在识别阳性和负性RNA病毒的宿主因素方面被证明是有效的.

结论:

基于病毒复制的CRISPR KO查是一种有效和多功能的方法,用于识别对各种病毒复制至关重要的宿主因素.
这种方法克服了活病毒查的局限性,并促进了新型宿主病毒相互作用的发现.
确定的宿主因素代表了开发宿主导抗病毒疗法和医疗对策的潜在新目标.

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