近日，以色列魏茨曼科学研究所Noam Stern-Ginossar等研究人员合作完成巨细胞病毒依赖性的病毒编码高分辨率筛选。该研究于2024年6月5日在线发表于国际一流学术期刊《自然》。

研究人员通过对人类巨细胞病毒进行工程改造，直接从病毒基因组表达单向导RNA文库，开发出了基于病毒编码CRISPR的直接读出筛选（VECOS），这是一种灵敏、多用途、以病毒为中心的方法，能以集合的形式分析病毒感染的不同阶段。利用这种方法，研究人员确定了数百种宿主依赖和限制因子，并量化了它们对病毒基因组复制、病毒颗粒分泌和分泌颗粒感染性的直接影响，为病毒与宿主的相互作用提供了一个多维视角。

这些高分辨率测量结果表明，改变人类巨细胞病毒（HCMV）生命周期晚期阶段的扰动主要调节病毒颗粒的质量而非数量，从而将正确的病毒组装确定为严重依赖病毒-宿主相互作用的关键阶段。总之，VECOS有助于系统地高分辨率分析人类蛋白质在感染周期中的作用，为深入研究宿主与疱疹病毒的相互作用提供了路线图。

研究人员表示，基因筛选改变了人们探究病毒感染对细胞因子要求的能力，但目前大多数方法的灵敏度有限，偏重于感染的早期阶段，而且只能提供简单的表型信息，这些信息往往基于感染细胞的存活率。

附：英文原文

Title: A virally encoded high-resolution screen of cytomegalovirus dependencies

Author: Finkel, Yaara, Nachshon, Aharon, Aharon, Einav, Arazi, Tamar, Simonovsky, Elena, Dobeov, Martina, Saud, Zack, Gluck, Avi, Fisher, Tal, Stanton, Richard J., Schwartz, Michal, Stern-Ginossar, Noam

Issue&Volume: 2024-06-05

Abstract: Genetic screens have transformed our ability to interrogate cellular factor requirements for viral infections1,2, but most current approaches are limited in their sensitivity, biased towards early stages of infection and provide only simplistic phenotypic information that is often based on survival of infected cells2,3,4. Here, by engineering human cytomegalovirus to express single guide RNA libraries directly from the viral genome, we developed virus-encoded CRISPR-based direct readout screening (VECOS), a sensitive, versatile, viral-centric approach that enables profiling of different stages of viral infection in a pooled format. Using this approach, we identified hundreds of host dependency and restriction factors and quantified their direct effects on viral genome replication, viral particle secretion and infectiousness of secreted particles, providing a multi-dimensional perspective on virus–host interactions. These high-resolution measurements reveal that perturbations altering late stages in the life cycle of human cytomegalovirus (HCMV) mostly regulate viral particle quality rather than quantity, establishing correct virion assembly as a critical stage that is heavily reliant on virus–host interactions. Overall, VECOS facilitates systematic high-resolution dissection of the role of human proteins during the infection cycle, providing a roadmap for in-depth study of host–herpesvirus interactions.

DOI: 10.1038/s41586-024-07503-z

Source: https://www.nature.com/articles/s41586-024-07503-z