Specially Appointed Assistant Professor Okayama University Okayama, Japan
Tomoaki Mori (Okayama University)| Kento Nakamura (Okayama University)| Keisuke Masaoka (Okayama University)| Koichi Mori (Okayama University)| Takamasa Tobimatsu (Okayama University)| Takashi Sera (Okayama University)
Previously, we reported that our designed artificial DNA-cleaving enzyme which comprises our artificial DNA-binding protein and a DNA-cleaving domain inhibited DNA replication of human papilloma virus (HPV) in mammalian cells by cleaving its target HPV ori plasmid sequence-specifically. Our artificial DNA-binding proteins have much higher affinity and selectivity than other DNA-binding proteins such as CRISPR-dCas9. In addition, our artificial DNA-cleaving enzymes cleave target DNA with multiple turnovers while CRISPR acts as a single-turnover enzyme. Thereafter, we applied this methodology for inactivating DNA viruses to RNA viruses. We have recently developed artificial RNA-binding proteins with higher affinity and selectivity than other RNA-binding proteins such as CRISPR-Cas13. In the present study, in order to inhibit RNA replication of influenza virus, we developed artificial RNA-cleaving enzymes which comprise our artificial RNA-binding protein and an RNA-cleaving domain. After we confirmed that an artificial RNA-binding protein specifically bound to its target RNA, we demonstrated that the artificial RNA-cleaving enzymes site-selectively and efficiently cleaved their target RNA sequence in an influenza viral genome in vitro. And we also demonstrated that the artificial RNA-cleaving enzyme effectively inhibited RNA replication of influenza virus in mammalian cells due to cleavage of target RNA. We will present the detail in this conference.