Development and Characterization of a Reverse Genetics System for a Human-Derived Severe Fever With Thrombocytopenia Syndrome Virus Isolate From South Korea

Severe fever with thrombocytopenia syndrome virus (SFTSV) is an emerging, tick-borne Bandavirus that causes lethal disease in humans. As there are no licensed vaccines and therapeutics for SFTSV, there is an urgent need to develop countermeasures against it. In this respect, a reverse genetics (RG)...

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Autores principales:	Seok-Min Yun, Tae-Young Lee, Hee-Young Lim, Jungsang Ryou, Joo-Yeon Lee, Young-Eui Kim
Formato:	article
Lenguaje:	EN
Publicado:	Frontiers Media S.A. 2021
Materias:	severe fever with thrombocytopenia syndrome virus reverse genetics system SFTSV animal model recombinant virus Microbiology QR1-502
Acceso en línea:	https://doaj.org/article/9c032f05f6214ea89a9c20197e69da1b
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spelling	oai:doaj.org-article:9c032f05f6214ea89a9c20197e69da1b2021-11-11T09:21:22ZDevelopment and Characterization of a Reverse Genetics System for a Human-Derived Severe Fever With Thrombocytopenia Syndrome Virus Isolate From South Korea1664-302X10.3389/fmicb.2021.772802https://doaj.org/article/9c032f05f6214ea89a9c20197e69da1b2021-11-01T00:00:00Zhttps://www.frontiersin.org/articles/10.3389/fmicb.2021.772802/fullhttps://doaj.org/toc/1664-302XSevere fever with thrombocytopenia syndrome virus (SFTSV) is an emerging, tick-borne Bandavirus that causes lethal disease in humans. As there are no licensed vaccines and therapeutics for SFTSV, there is an urgent need to develop countermeasures against it. In this respect, a reverse genetics (RG) system is a powerful tool to help achieve this goal. Herein, we established a T7 RNA polymerase-driven RG system to rescue infectious clones of a Korean SFTSV human isolate entirely from complementary DNA (cDNA). To establish this system, we cloned cDNAs encoding the three antigenomic segments into transcription vectors, with each segment transcribed under the control of the T7 promoter and the hepatitis delta virus ribozyme (HdvRz) sequences. We also constructed two helper plasmids expressing the nucleoprotein (NP) or viral RNA-dependent RNA polymerase (RdRp) under the control of the T7 promoter and the encephalomyocarditis virus (EMCV) internal ribosome entry site (IRES). After co-transfection into BHK/T7-9 cells with three transcription and two helper plasmids, then passaging in Vero E6 or Huh-7 cells, we confirmed efficient rescue of the recombinant SFTSV. By evaluating the in vitro and in vivo virological properties of the parental and rescued SFTSVs, we show that the rescued virus exhibited biological properties similar to those of the parental virus. This system will be useful for identifying molecular viral determinants of SFTSV infection and pathogenesis and for facilitating the development of vaccine and antiviral approaches.Seok-Min YunTae-Young LeeHee-Young LimJungsang RyouJoo-Yeon LeeYoung-Eui KimFrontiers Media S.A.articlesevere fever with thrombocytopenia syndrome virusreverse genetics systemSFTSVanimal modelrecombinant virusMicrobiologyQR1-502ENFrontiers in Microbiology, Vol 12 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	severe fever with thrombocytopenia syndrome virus reverse genetics system SFTSV animal model recombinant virus Microbiology QR1-502
spellingShingle	severe fever with thrombocytopenia syndrome virus reverse genetics system SFTSV animal model recombinant virus Microbiology QR1-502 Seok-Min Yun Tae-Young Lee Hee-Young Lim Jungsang Ryou Joo-Yeon Lee Young-Eui Kim Development and Characterization of a Reverse Genetics System for a Human-Derived Severe Fever With Thrombocytopenia Syndrome Virus Isolate From South Korea
description	Severe fever with thrombocytopenia syndrome virus (SFTSV) is an emerging, tick-borne Bandavirus that causes lethal disease in humans. As there are no licensed vaccines and therapeutics for SFTSV, there is an urgent need to develop countermeasures against it. In this respect, a reverse genetics (RG) system is a powerful tool to help achieve this goal. Herein, we established a T7 RNA polymerase-driven RG system to rescue infectious clones of a Korean SFTSV human isolate entirely from complementary DNA (cDNA). To establish this system, we cloned cDNAs encoding the three antigenomic segments into transcription vectors, with each segment transcribed under the control of the T7 promoter and the hepatitis delta virus ribozyme (HdvRz) sequences. We also constructed two helper plasmids expressing the nucleoprotein (NP) or viral RNA-dependent RNA polymerase (RdRp) under the control of the T7 promoter and the encephalomyocarditis virus (EMCV) internal ribosome entry site (IRES). After co-transfection into BHK/T7-9 cells with three transcription and two helper plasmids, then passaging in Vero E6 or Huh-7 cells, we confirmed efficient rescue of the recombinant SFTSV. By evaluating the in vitro and in vivo virological properties of the parental and rescued SFTSVs, we show that the rescued virus exhibited biological properties similar to those of the parental virus. This system will be useful for identifying molecular viral determinants of SFTSV infection and pathogenesis and for facilitating the development of vaccine and antiviral approaches.
format	article
author	Seok-Min Yun Tae-Young Lee Hee-Young Lim Jungsang Ryou Joo-Yeon Lee Young-Eui Kim
author_facet	Seok-Min Yun Tae-Young Lee Hee-Young Lim Jungsang Ryou Joo-Yeon Lee Young-Eui Kim
author_sort	Seok-Min Yun
title	Development and Characterization of a Reverse Genetics System for a Human-Derived Severe Fever With Thrombocytopenia Syndrome Virus Isolate From South Korea
title_short	Development and Characterization of a Reverse Genetics System for a Human-Derived Severe Fever With Thrombocytopenia Syndrome Virus Isolate From South Korea
title_full	Development and Characterization of a Reverse Genetics System for a Human-Derived Severe Fever With Thrombocytopenia Syndrome Virus Isolate From South Korea
title_fullStr	Development and Characterization of a Reverse Genetics System for a Human-Derived Severe Fever With Thrombocytopenia Syndrome Virus Isolate From South Korea
title_full_unstemmed	Development and Characterization of a Reverse Genetics System for a Human-Derived Severe Fever With Thrombocytopenia Syndrome Virus Isolate From South Korea
title_sort	development and characterization of a reverse genetics system for a human-derived severe fever with thrombocytopenia syndrome virus isolate from south korea
publisher	Frontiers Media S.A.
publishDate	2021
url	https://doaj.org/article/9c032f05f6214ea89a9c20197e69da1b
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Development and Characterization of a Reverse Genetics System for a Human-Derived Severe Fever With Thrombocytopenia Syndrome Virus Isolate From South Korea

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