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

Area of Science:

• Virology
• Genetics
• Molecular Biology

Background:

• Discovering cellular genes crucial for virus entry is a key challenge in virology.
• Genome-wide loss-of-function screens identify necessary genes but are hindered by genetic redundancy.

Purpose of the Study:

• To develop and validate a genome-wide CRISPR activation screening strategy.
• To identify single genes sufficient for viral entry into normally uninfectable cells.

Main Methods:

• Utilized a genome-wide CRISPR activation screen.
• Employed sequential rounds of viral infection to enhance screening sensitivity.
• Applied the strategy to identify entry factors for Ebola and rabies viruses.

Main Results:

• The CRISPR activation screen successfully identified genes sufficient for viral entry.
• Sequential infection rounds significantly improved screening sensitivity.
• The method proved generalizable to distinct viruses, including Ebola and rabies.

Conclusions:

• CRISPR activation screening is a powerful tool for discovering viral entry factors.
• This strategy overcomes genetic redundancy issues inherent in loss-of-function screens.
• The approach has broad applicability for accelerating the discovery of host factors mediating viral entry.