title: Flavivirus reverse genetic systems, construction techniques and applications: a historical perspective.
abstract: The study of flaviviruses, which cause some of the most important emerging tropical and sub-tropical human arbovirus diseases, has greatly benefited from the use of reverse genetic systems since its first development for yellow fever virus in 1989. Reverse genetics technology has completely revolutionized the study of these viruses, making it possible to manipulate their genomes and evaluate the direct effects of these changes on their biology and pathogenesis. The most commonly used reverse genetics system is the infectious clone technology. Whilst flavivirus infectious clones provide a powerful tool, their construction as full-length cDNA molecules in bacterial vectors can be problematic, laborious and time consuming, because they are often unstable, contain unwanted induced substitutions and may be toxic for bacteria due to viral protein expression. The incredible technological advances that have been made during the past 30years, such as the use of PCR or new sequencing methods, have allowed the development of new approaches to improve preexisting systems or elaborate new strategies that overcome these problems. This review summarizes the evolution and major technical breakthroughs in the development of flavivirus reverse genetics technologies and their application to the further understanding and control of these viruses and their diseases.
authors.affiliation: Aix Marseille Université, IRD French Institute of Research for Development, EHESP French School of Public Health, EPV UMR_D 190 "Emergence des Pathologies Virales", 13385 Marseille, France. Electronic address: fab1.aubry@gmail.com.
authors.forename: Fabien
authors.initials: F
authors.lastname: Aubry
authors.affiliation: Aix Marseille Université, IRD French Institute of Research for Development, EHESP French School of Public Health, EPV UMR_D 190 "Emergence des Pathologies Virales", 13385 Marseille, France; Institut Hospitalo-Universitaire Méditerranée Infection, Marseille, France.
authors.forename: Antoine
authors.initials: A
authors.lastname: Nougairède
authors.affiliation: Aix Marseille Université, IRD French Institute of Research for Development, EHESP French School of Public Health, EPV UMR_D 190 "Emergence des Pathologies Virales", 13385 Marseille, France.
authors.forename: Ernest A
authors.initials: EA
authors.lastname: Gould
authors.affiliation: Aix Marseille Université, IRD French Institute of Research for Development, EHESP French School of Public Health, EPV UMR_D 190 "Emergence des Pathologies Virales", 13385 Marseille, France; Institut Hospitalo-Universitaire Méditerranée Infection, Marseille, France.
authors.forename: Xavier
authors.initials: X
authors.lastname: de Lamballerie
mesh_terms: D003001:Cloning, Molecular; D018076:DNA, Complementary; D005416:Flavivirus / Q000235:genetics* / Q000276:immunology / Q000502:physiology; D005822:Genetic Vectors; D012367:RNA, Viral / Q000235:genetics; D059386:Reverse Genetics; D014765:Viral Vaccines / Q000235:genetics; D014779:Virus Replication / Q000235:genetics
chemical_list: D018076:DNA, Complementary; D012367:RNA, Viral; D014765:Viral Vaccines
keywords: Flavivirus; Infectious clone; Reverse genetics