The present study investigated the role of interferon-inducible pathways in herpes simplex virus type 1-infected mice transduced with an adenoviral vector expressing murine interferon-β (Ad:IFN-β). Wild type mice or RNase L −/− mice deficient in responses to 2′-5′ oligoadenylate synthetase activation, or lacking RNA-dependent protein kinase and transduced with Ad:IFN-β showed enhanced survival following HSV-1 infection. The protective effect was associated with a reduction in viral gene expression in the cornea and trigeminal ganglion in wild type mice as well as the trigeminal ganglion of RNase L −/− mice. However, the efficacy of Ad:IFN-β was lost in the corneas of RNase L −/− mice and significantly diminished in both the cornea and trigeminal ganglion as measured by viral gene expression in RNA-dependent protein kinase deficient mice. Collectively, the data suggest survival rates of viral-infected mice do not reflect the replication capacity as measured by herpes simplex virus type one lytic gene expression.