title: Utilisation of ISA Reverse Genetics and Large-Scale Random Codon Re-Encoding to Produce Attenuated Strains of Tick-Borne Encephalitis Virus within Days.
abstract: Large-scale codon re-encoding is a new method of attenuating RNA viruses. However, the use of infectious clones to generate attenuated viruses has inherent technical problems. We previously developed a bacterium-free reverse genetics protocol, designated ISA, and now combined it with large-scale random codon-re-encoding method to produce attenuated tick-borne encephalitis virus (TBEV), a pathogenic flavivirus which causes febrile illness and encephalitis in humans. We produced wild-type (WT) and two re-encoded TBEVs, containing 273 or 273+284 synonymous mutations in the NS5 and NS5+NS3 coding regions respectively. Both re-encoded viruses were attenuated when compared with WT virus using a laboratory mouse model and the relative level of attenuation increased with the degree of re-encoding. Moreover, all infected animals produced neutralizing antibodies. This novel, rapid and efficient approach to engineering attenuated viruses could potentially expedite the development of safe and effective new-generation live attenuated vaccines.
authors.affiliation: UMR "Emergence des Pathologies Virales" (EPV: Aix-Marseille University-IRD 190-Inserm 1207-EHESP), Marseille, France.
authors.forename: Lauriane
authors.initials: L
authors.lastname: de Fabritus
authors.affiliation: UMR "Emergence des Pathologies Virales" (EPV: Aix-Marseille University-IRD 190-Inserm 1207-EHESP), Marseille, France.|Institut Hospitalo-Universitaire Méditerranée Infection, Marseille, France.
authors.forename: Antoine
authors.initials: A
authors.lastname: Nougairède
authors.affiliation: UMR "Emergence des Pathologies Virales" (EPV: Aix-Marseille University-IRD 190-Inserm 1207-EHESP), Marseille, France.
authors.forename: Fabien
authors.initials: F
authors.lastname: Aubry
authors.affiliation: UMR "Emergence des Pathologies Virales" (EPV: Aix-Marseille University-IRD 190-Inserm 1207-EHESP), Marseille, France.
authors.forename: Ernest A
authors.initials: EA
authors.lastname: Gould
authors.affiliation: UMR "Emergence des Pathologies Virales" (EPV: Aix-Marseille University-IRD 190-Inserm 1207-EHESP), Marseille, France.|Institut Hospitalo-Universitaire Méditerranée Infection, Marseille, France.
authors.forename: Xavier
authors.initials: X
authors.lastname: de Lamballerie
mesh_terms: D000818:Animals; D057134:Antibodies, Neutralizing / Q000096:biosynthesis*; D000914:Antibodies, Viral / Q000096:biosynthesis*; D002460:Cell Line; D003062:Codon / Q000737:chemistry* / Q000378:metabolism; D006224:Cricetinae; D004669:Encephalitis Viruses, Tick-Borne / Q000187:drug effects / Q000235:genetics / Q000276:immunology*; D004675:Encephalitis, Tick-Borne / Q000276:immunology / Q000401:mortality / Q000517:prevention & control* / Q000821:virology; D005260:Female; D006801:Humans; D051379:Mice; D008810:Mice, Inbred C57BL; D009154:Mutation; D016366:Open Reading Frames; D020365:RNA Helicases / Q000235:genetics / Q000276:immunology; D059386:Reverse Genetics / Q000379:methods*; D012697:Serine Endopeptidases / Q000235:genetics / Q000276:immunology; D016019:Survival Analysis; D014613:Vaccines, Attenuated; D017361:Viral Nonstructural Proteins / Q000235:genetics / Q000276:immunology; D014765:Viral Vaccines / Q000008:administration & dosage / Q000235:genetics* / Q000276:immunology; D043583:Nucleoside-Triphosphatase; D053487:DEAD-box RNA Helicases; D000085242:Viral Proteases
chemical_list: D057134:Antibodies, Neutralizing; D000914:Antibodies, Viral; D003062:Codon; D020365:RNA Helicases; D012697:Serine Endopeptidases; D014613:Vaccines, Attenuated; D017361:Viral Nonstructural Proteins; D014765:Viral Vaccines; C060028:NS3 protein, flavivirus; D043583:Nucleoside-Triphosphatase; D053487:DEAD-box RNA Helicases; D000085242:Viral Proteases; C096041:NS5 protein, flavivirus