A virally encoded high-resolution screen of cytomegalovirus dependencies

Finkel, Yaara, Nachshon, Aharon, Aharon, Einav, Arazi, Tamar, Simonovsky, Elena, Dobesová, Martina, Saud, Zack, Gluck, Avi, Fisher, Tal, Stanton, Richard J. ORCID: https://orcid.org/0000-0002-6799-1182, Schwartz, Michal and Stern-Ginossar, Noam 2024. A virally encoded high-resolution screen of cytomegalovirus dependencies. Nature 630 , pp. 712-719. 10.1038/s41586-024-07503-z

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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.

Item Type:	Article
Date Type:	Publication
Status:	Published
Schools:	Medicine Systems Immunity Research Institute (SIURI)
Publisher:	Nature Research
ISSN:	0028-0836
Funders:	MRC
Date of First Compliant Deposit:	25 June 2024
Date of Acceptance:	1 May 2024
Last Modified:	09 Nov 2024 18:30
URI:	https://orca.cardiff.ac.uk/id/eprint/169512

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