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Open AccessArticle

Novel Toscana Virus Reverse Genetics System Establishes NSs as an Antagonist of Type I Interferon Responses

1
CellNetworks Cluster of Excellence, University Hospital Heidelberg, 69120 Heidelberg, Germany
2
Center for Integrative Infectious Diseases Research (CIID), Virology, University Hospital Heidelberg, 69120 Heidelberg, Germany
3
Wageningen Bioveterinary Research, Department of Virology, 8221 RA Lelystad, The Netherlands
4
Center for Integrative Infectious Diseases Research (CIID), Molecular Virology, University Hospital Heidelberg, 69120 Heidelberg, Germany
5
Structural Virology Unit, Pasteur Institute, 75015 Paris, France
6
Laboratory of Virology, Wageningen University, 6708 PB Wageningen, The Netherlands
7
INRAE, EPHE, Viral Infections and Comparative Pathology (IVPC), University Claude Bernard Lyon1, University of Lyon, UMR754, 69007 Lyon, France
*
Authors to whom correspondence should be addressed.
These authors contributed equally to this work.
Viruses 2020, 12(4), 400; https://doi.org/10.3390/v12040400
Received: 4 March 2020 / Revised: 30 March 2020 / Accepted: 3 April 2020 / Published: 4 April 2020
(This article belongs to the Special Issue Emerging Arboviruses)
The sand fly-borne Toscana virus (TOSV) is the major cause of human meningoencephalitis in the Mediterranean basin during the summer season. In this work, we have developed a T7 RNA polymerase-driven reverse genetics system to recover infectious particles of a lineage B strain of TOSV. The viral protein pattern and growth properties of the rescued virus (rTOSV) were found to be similar to those of the corresponding wild-type (wt) virus. Using this system, we genetically engineered a TOSV mutant lacking expression of the non-structural protein NSs (rTOSVɸNSs). Unlike rTOSV and the wt virus, rTOSVɸNSs was unable to (i) suppress interferon (IFN)-b messenger RNA induction; and (ii) grow efficiently in cells producing IFN-b. Together, our results highlight the importance of NSs for TOSV in evading the IFN response and provide a comprehensive toolbox to investigate the TOSV life cycle in mammalian and insect host cells, including several novel polyclonal antibodies. View Full-Text
Keywords: Arbovirus; Bunyavirales; interferon; neglected diseases; Phenuiviridae; Phlebovirus; reverse genetics; sand fly fever; Toscana virus Arbovirus; Bunyavirales; interferon; neglected diseases; Phenuiviridae; Phlebovirus; reverse genetics; sand fly fever; Toscana virus
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Woelfl, F.; Léger, P.; Oreshkova, N.; Pahmeier, F.; Windhaber, S.; Koch, J.; Stanifer, M.; Roman Sosa, G.; Uckeley, Z.M.; Rey, F.A.; Boulant, S.; Kortekaas, J.; Wichgers Schreur, P.J.; Lozach, P.-Y. Novel Toscana Virus Reverse Genetics System Establishes NSs as an Antagonist of Type I Interferon Responses. Viruses 2020, 12, 400.

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