Inhibition of human papillomavirus replication by using artificial zinc-finger nucleases.

Recently, we have designed artificial zinc-finger proteins (AZPs) that prevent a viral replication protein, E2, of human papillomavirus type 18 (HPV-18) from binding to its replication origin and demonstrated that the gene-delivered AZPs inhibited HPV-18 DNA replication in mammalian cells. In the present study, we examined a new approach to inhibition of DNA virus replication by using an AZP-nuclease fusion. In transient replication assays for HPV-18, the gene-delivered AZP-nuclease fusion reduced the viral DNA replication rate significantly. Moreover, it was demonstrated by ligation-mediated PCR that viral DNA regions close to the AZP-binding site were cleaved in the cells by the AZP-nuclease. Thus, our results demonstrate that AZP-nucleases have potentials to inhibit replication of any DNA viruses whose replication mechanisms remain unsolved.