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Nucleic Acids Symposium Series 2006 50(1):313-314; doi:10.1093/nass/nrl156
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© 2006 Oxford University Press

Application of artificial zinc-finger proteins to inhibition of DNA replication of human papillomavirus

Takashi Mino, Tomoaki Mori, Naoki Matsumoto, Yusuke Mineta, Tomoyuki Okamoto, Yasuhiro Aoyama and Takashi Sera

Department of Synthetic Chemistry and Biological Chemistry, Graduate School of Engineering, Kyoto University, Kyotodaigaku-Katsura, Nishikyo-ku, Kyoto 615-8510, Japan

Recently, we have demonstrated that plant DNA virus replication could be inhibited in Arabidopsis thaliana by using an artificial zinc-finger protein (AZP) and created AZP-based transgenic A. thaliana resistant to DNA virus infection. Here we apply the AZP technology to inhibition of replication of a mammalian DNA virus, human papillomavirus (HPV) type 18. Two AZPs, designated AZPHPV-1 and AZPHPV-2, were designed to block binding of the HPV-18 E2 replication protein to the replication origin. Both the designed AZPs had much higher affinities towards the replication origin than did the E2 protein, and efficiently blocked E2 binding in vitro. In transient replication assays, both AZPs inhibited the viral DNA replication: AZPHPV-2, especially, reduced the replication level to approximately 10%. Thus, it was demonstrated that the AZP technology could be applied not only to plant DNA viruses, but also to mammalian DNA viruses.


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