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.
Mino
Takashi
T
Department of Synthetic Chemistry and Biological Chemistry, Graduate School of Engineering, Kyoto University, Kyotodaigaku-Katsura, Nishikyo-ku, Kyoto 615-8510, Japan.
Mori
Tomoaki
T
Aoyama
Yasuhiro
Y
Sera
Takashi
T
D004261:DNA Replication; D004279:DNA, Viral / Q000096:biosynthesis*; D003851:Deoxyribonucleases / Q000235:genetics / Q000378:metabolism*; D052161:Human papillomavirus 18 / Q000235:genetics*; D009856:Oncogene Proteins, Viral / Q000037:antagonists & inhibitors; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D016335:Zinc Fingers
D004279:DNA, Viral; D009856:Oncogene Proteins, Viral; D011993:Recombinant Fusion Proteins; C069102:oncogene protein E2, Human papillomavirus type 18; D003851:Deoxyribonucleases