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Elucidating genes sufficient for viral entry into cells through sequential genome-wide CRISPR activation screens

Chai, T.; Wong, A.; Yin, Q.; von Creytz, I.; Weissman, J. S.; Saunders, R. A.; Prescott, J. B.; Loh, K. M.

2026-03-06 microbiology
10.64898/2026.03.06.710083 bioRxiv
Show abstract

A preeminent goal of virology is to discover cellular genes that mediate virus entry. Genome-wide loss-of-function screens can illuminate single genes necessary for virus entry, but are stymied by genetic redundancy. Here we report a genome-wide CRISPR activation screening strategy to discover single genes that are sufficient for viral entry into normally-uninfectable cells. Sequential rounds of viral infection vastly enhanced screening sensitivity. This sequential screening strategy was generalizable to two unrelated viruses--Ebola and rabies viruses--and could broadly accelerate the discovery of viral entry factors.

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