Alexander, A. J.T. et al. (2020) Development of a reverse genetics system for Toscana virus (lineage A). Viruses, 12(4), 411. (doi: 10.3390/v12040411)
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211590.pdf - Published Version Available under License Creative Commons Attribution. 2MB |
Abstract
Toscana virus (TOSV) is a Phlebovirus in the Phenuiviridae family, order Bunyavirales, found in the countries surrounding the Mediterranean. TOSV is an important cause of seasonal acute meningitis and encephalitis within its range. Here, we determined the full sequence of the TOSV strain 1500590, a lineage A virus obtained from an infected patient (Marseille, 2007) and used this in combination with other sequence information to construct functional cDNA plasmids encoding the viral L, M, and S antigenomic sequences under the control of the T7 RNA promoter to recover recombinant viruses. Importantly, resequencing identified two single nucleotide changes to a TOSV reference genome, which, when corrected, restored functionality to the polymerase L and made it possible to recover infectious recombinant TOSV (rTOSV) from cDNA, as well as establish a minigenome system. Using reverse genetics, we produced an NSs-deletant rTOSV and also obtained viruses expressing reporter genes instead of NSs. The availability of such a system assists investigating questions that require genetic manipulation of the viral genome, such as investigations into replication and tropism, and beyond these fundamental aspects, also the development of novel vaccine design strategies.
| Item Type: | Articles |
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| Additional Information: | Funding: This project received funding from the European Union’s Horizon 2020 Research and Innovation Programme Infravec2, Research infrastructures for the control of insect vector-borne diseases, under the grant agreement No 731060 (AK). It was also supported by the UK MRC (MC_UU_12014/8) (AK), the Fondation FINOVI, the Ecole Pratique des Hautes Etudes, the Institut National de la Recherche Agronomique, and the University of Lyon 1 (MR, FA). |
| Status: | Published |
| Refereed: | Yes |
| Glasgow Author(s) Enlighten ID: | Mair, Mr Daniel and Wilkie, Dr Gavin and Da Silva Filipe, Dr Ana and Brennan, Dr Benjamin and Alexander, Dr Akira and Dunlop, Dr James and Vattipally, Dr Sreenu and Kuchi, Dr Srikeerthana and Kohl, Professor Alain |
| Creator Roles: | Alexander, A.Conceptualization, Data curation, Formal analysis, Investigation, Methodology, Validation, Visualization, Writing – original draft, Writing – review and editing Kohl, A.Conceptualization, Data curation, Formal analysis, Funding acquisition, Investigation, Methodology, Project administration, Resources, Supervision, Writing – original draft, Writing – review and editing Kuchi, S.Data curation, Formal analysis, Investigation, Methodology, Software, Writing – review and editing Brennan, B.Formal analysis, Investigation, Methodology, Writing – original draft, Writing – review and editing Vattipally, S.Formal analysis, Investigation, Methodology, Software, Writing – review and editing Dunlop, J.Investigation, Methodology, Writing – review and editing Wilkie, G.Investigation, Methodology, Writing – review and editing Mair, D.Investigation, Methodology, Writing – review and editing Da Silva Filipe, A.Investigation, Methodology, Resources, Writing – review and editing |
| Authors: | Alexander, A. J.T., Confort, M.-P., Desloire, S., Dunlop, J. I., Kuchi, S., Sreenu, V. B., Mair, D., Wilkie, G. S., Da Silva Filipe, A., Brennan, B., Ratinier, M., Arnaud, F., and Kohl, A. |
| College/School: | College of Medical Veterinary and Life Sciences > School of Infection & Immunity College of Medical Veterinary and Life Sciences > School of Infection & Immunity > Centre for Virus Research |
| Journal Name: | Viruses |
| Publisher: | MDPI |
| ISSN: | 1999-4915 |
| ISSN (Online): | 1999-4915 |
| Copyright Holders: | Copyright © 2020 by the authors |
| First Published: | First published in Viruses 12(4):411 |
| Publisher Policy: | Reproduced under a Creative Commons license |
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