A range of NRF2-driven Restriction Factors inhibit Hepatitis C virus Replication

The transcription factor NRF2 (Nuclear factor erythroid 2-related factor 2) has recently emerged as an important component in non-canonical anti-viral immunity. We have previously reported that NRF2 agonists induce a distinct interferon (IFN)-independent antiviral program that is broadly effective in limiting virus replication of human pathogenic viruses, including SARS-CoV-2. Using both live infectious virus as well as a subgenomic replicon system, we demonstrate that CRISPR-mediated induction of endogenous NRF2 expression restricted Hepatitis C virus (HCV) replication. Further, using a focused CRISPR-activation (CRISPRa) screen, we identified a broad range of NRF2-driven genes with antiviral activity against HCV. Among these, the RNA-binding protein ZC3H12C exhibited potent antiviral effects across multiple assays, identifying ZC3H12C as a previously unrecognized HCV restriction factor. In conclusion, we demonstrate that NRF2 controls a range of restriction factors that can limit HCV replication and identify ZC3H12C as a novel antiviral effector gene.