Development of a reverse genetics system for Toscana virus (lineage A)

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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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
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:Wilkie, Dr Gavin and Kuchi, Dr Srikeerthana and Alexander, Dr Akira and Dunlop, Dr James and Da Silva Filipe, Dr Ana and Vattipally, Dr Sreenu and Brennan, Dr Benjamin and Mair, Mr Daniel and Kohl, Professor Alain
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 > Institute of Infection Immunity and Inflammation
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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Project CodeAward NoProject NamePrincipal InvestigatorFunder's NameFunder RefLead Dept
MC_UU_12014/8
173819INFRAVEC-2Alain KohlEuropean Commission (EC)Kohl, Dr AlainIII-MRC-GU Centre for Virus Research