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Cleavage of Influenza RNA Using Artificial RNA‐cleaving Enzyme
Author(s) -
Mori Tomoaki,
Nakamura Kento,
Masaoka Keisuke,
Mori Koichi,
Tobimatsu Takamasa,
Sera Takashi
Publication year - 2021
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.2021.35.s1.01768
Subject(s) - rna , dna , rna dependent rna polymerase , biology , rna binding protein , microbiology and biotechnology , biochemistry , chemistry , gene
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.