Journal of Antivirals & Antiretrovirals
Open Access
ISSN: 1948-5964
+44 1300 500008
ISSN: 1948-5964
+44 1300 500008
Takashi Sera
Okayama University, Japan
Posters & Accepted Abstracts: J Antivir Antiretrovir
Previously, we reported that artificial zinc-finger proteins (AZPs) inhibited virus DNA replication in planta and in mammalian cells by blocking binding of a viral replication protein to its replication origin. However, the replication mechanisms of viruses of interest need to be disentangled for the application. To develop more widely applicable methods for antiviral therapy, we explored the feasibility of inhibition of HPV-18 replication as a model system by cleaving its viral genome. To this end, we fused a nuclease cleaving DNA as a monomer to an AZP that binds to the viral genome. The resulting artificial endonuclease cleaved its target DNA plasmid efficiently and sequence specifically in vitro. Then, we confirmed that transfection with a plasmid expressing the artificial endonuclease inhibited HPV-18 DNA replication in transient replication assays using mammalian cells. Linker mediated PCR analysis revealed that the artificial endonuclease cleaved an HPV-18 ori plasmid around its binding site. Finally, we demonstrated that the protein delivered artificial endonuclease inhibited HPV-18 DNA replication as well and did not show any significant cytotoxicity. Thus, both gene and protein delivered zinc finger based artificial endonucleases efficiently inhibited HPV-18 DNA replication, leading to development of a more universal antiviral therapy for human DNA viruses.
Email: sera@cc.okayama-u.ac.jp