Inhibition of VCP by chemical inhibitors decreased WNV infection in a dose-dependent manner. Knockdown of endogenous VCP level using siRNA suppressed WNV infection. Depletion of VCP levels suppressed WNV infection at the early stages of WNV replication cycle. Depletion of VCP levels lowered nascent WNV genomic RNA. VCP participates in early stages and viral genomic RNA replication.