Development of a Reverse Genetics System for Toscana Virus (Lineage A)

Autor:	Daniel Mair, Srikeerthana Kuchi, Sophie Desloire, James I. Dunlop, Marie-Pierre Confort, Benjamin Brennan, Alain Kohl, Gavin S. Wilkie, Vattipally B. Sreenu, Maxime Ratinier, Frederick Arnaud, Ana da Silva Filipe, Akira J T Alexander
Přispěvatelé:	Infections Virales et Pathologie Comparée - UMR 754 (IVPC), École pratique des hautes études (EPHE), Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-Université Claude Bernard Lyon 1 (UCBL), Université de Lyon-Université de Lyon-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), Université Claude Bernard Lyon 1 (UCBL), Université de Lyon, University of Glasgow, MRC - University of Glasgow Centre for Virus Research, Institut de biologie et chimie des protéines [Lyon] (IBCP), Université de Lyon-Université de Lyon-Centre National de la Recherche Scientifique (CNRS), European Union (EU)731060Medical Research Council UK (MRC)MC_UU_12014/8Fondation FINOVI Institut National de la Recherche Agronomique University of Lyon 1 Ecole Pratique des Hautes Etudes
Rok vydání:	2020
Předmět:	0301 basic medicine Phlebovirus [SDV]Life Sciences [q-bio] 030106 microbiology lcsh:QR1-502 Genome Viral Genome Virus Article lcsh:Microbiology Bunyavirales 03 medical and health sciences Viral Proteins reverse genetics Plasmid Virology Complementary DNA Humans Promoter Regions Genetic Toscana virus biology Sandfly fever Naples virus biology.organism_classification Reverse genetics 3. Good health 030104 developmental biology Infectious Diseases Phlebotomus Fever A549 Cells Reference genome
Zdroj:	Viruses Viruses, Vol 12, Iss 411, p 411 (2020) Volume 12 Issue 4 Viruses, MDPI, 2020, 12 (4), pp.411. ⟨10.3390/v12040411⟩
ISSN:	1999-4915
DOI:	10.3390/v12040411
Popis:	International audience; 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.
Databáze:	OpenAIRE
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