Inducible CRISPR activation screen for interferon-stimulated genes identifies OAS1 as a SARS-CoV-2 restriction factor.

Inducible CRISPR activation screen for interferon-stimulated genes identifies OAS1 as a SARS-CoV-2 restriction factor.

Blog Article

Interferons establish an antiviral state through the induction of hundreds of interferon-stimulated genes (ISGs).The mechanisms and viral specificities for most ISGs remain incompletely understood.To enable high-throughput interrogation of ISG antiviral functions in pooled genetic screens while mitigating potentially glass confounding effects of endogenous interferon and antiproliferative/proapoptotic ISG activities, we adapted a CRISPR-activation (CRISPRa) system for inducible ISG expression in isogenic cell lines with and without the capacity to respond to interferons.We used this platform to screen for ISGs that restrict SARS-CoV-2.

Results included ISGs previously described to restrict SARS-CoV-2 and novel candidate antiviral factors.We validated a subset of these by complementary CRISPRa and cDNA expression experiments.OAS1, a top-ranked hit Cilantro across multiple screens, exhibited strong antiviral effects against SARS-CoV-2, which required OAS1 catalytic activity.These studies demonstrate a high-throughput approach to assess antiviral functions within the ISG repertoire, exemplified by identification of multiple SARS-CoV-2 restriction factors.