The emergence of RNA interference (RNAi) as a powerful tool for silencing gene expression has spurred considerable interest in its experimental and therapeutic potential. RNAi is a cellular process of gene silencing in which small duplexes of RNA specifically target a homologous sequence for cleavage by cellular ribonucleases. The introduction of 21-23 nucleotide RNA duplexes, termed small interfering RNAs (siRNAs), into mammalian cells can specifically degrade homologous mRNAs. RNAi efficiently silences the expression of both cellular and viral RNAs. A number of groups have demonstrated that siRNAs interfere with hepatitis C virus (HCV) gene expression and replication. Additionally, cellular genes are efficiently silenced in the presence of replicating HCV. These studies lay the foundation for using RNAi as an experimental tool for studying HCV replication and defining host genes that are significant for viral replication. The potential for RNAi as an antiviral therapy remains less clear, as it will face many of the challenges that have hindered nucleic acid therapies in the past.