1. Development of an entirely cloned cDNA-based reverse genetics system for Tofla virus of orthonairovirus.
- Author
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Wulandari, Shelly, Nyampong, Samuel, Lokupathirage, Sithumini M.W., Yoshimatsu, Kumiko, Shimoda, Hiroshi, and Hayasaka, Daisuke
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RNA polymerases , *VIRAL genes , *VIRAL genetics , *COMPLEMENTARY DNA , *HEMORRHAGIC fever , *REVERSE genetics - Abstract
The genus Orthonairovirus includes highly pathogenic tick-borne viruses such as the Crimean-Congo hemorrhagic fever orthonairovirus (CCHFV). A reverse genetics system is an indispensable tool for determining the viral factors related to pathogenicity. Tofla orthonairovirus (TFLV) is a recently identified virus isolated from ticks in Japan and our research has suggested that TFLV is a useful model for studying pathogenic orthonairoviruses. In this study, we successfully established a reverse genetics system for TFLV using T7 RNA polymerase. Recombinant TFLV was generated by transfecting cloned complementary DNAs encoding the TFLV genome into BSR T7/5 cells expressing T7 RNA polymerase. We were able to rescue infectious recombinant TFLV mutant (rTFLVmt) and wild-type TFLV (rTFLVpt) viruses, which exhibited indistinguishable growth kinetics in mammalian cells and pathogenicity in A129 mice compared with the authentic virus. Our approach provides a valuable method for establishing reverse genetics system for orthonairoviruses. • T7 RNA polymerase-driven reverse genetic system of the Tofla orthonairovirus was developed. • Recombinant TFLVs showed similar virological characteristics with the parent virus. • This system provides an alternative method for reveres genetics of orthonairovirus. • TFLV recombinants harboring mutations can be generated. • This system is a useful tool for investigating viral genes responsible for pathogenicity. [ABSTRACT FROM AUTHOR]
- Published
- 2024
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