Interferon inhibits a model RNA virus via a limited set of inducible effector genes [ISG_CRISPR_Screen]

Host cells produce interferon (IFN) in response to viral infections. Secreted interferon results in the transcription and production of hundreds of interferon-stimulated genes (ISGs). An ISG-targeted CRISPR screen using IFN beta-treated U-2 OS cells was performed to determine which ISGs were required in order for host cells to suppress Venezuelan equine encephalitis virus (VEEV) infection. Human U-2 OS cells were transduced with a human interferon-stimulated gene CRISPR knockout library, selected for transduced cells, treated with IFNβ, infected with Venezuelan equine encephalitis virus encoding a GFP reporter, and GFP-positive cells that became infected despite IFNβ suppression were collected by FACS. The screen was performed as 3 independent biological replicates at approximately 1270x, 430x, and 1160x library coverage.