title: A Full-Length Infectious cDNA Clone of Zika Virus from the 2015 Epidemic in Brazil as a Genetic Platform for Studies of Virus-Host Interactions and Vaccine Development.
abstract: An arthropod-borne virus, Zika virus (ZIKV), has recently emerged as a major human pathogen. Associated with complications during perinatal development and Guillain-Barré syndrome in adults, ZIKV raises new challenges for understanding the molecular determinants of flavivirus pathogenesis. This underscores the necessity for the development of a reverse genetic system based on an epidemic ZIKV strain. Here, we describe the generation and characterization in cell cultures of an infectious cDNA clone of ZIKV isolated from the 2015 epidemic in Brazil. The cDNA-derived ZIKV replicated efficiently in a variety of cell lines, including those of both neuronal and placental origin. We observed that the growth of cDNA-derived virus was attenuated compared to the growth of the parental isolate in most cell lines, which correlates with substantial differences in sequence heterogeneity between these viruses that were determined by deep-sequencing analysis. Our findings support the role of genetic diversity in maintaining the replicative fitness of viral populations under changing conditions. Moreover, these results indicate that caution should be exercised when interpreting the results of reverse-genetics experiments in attempts to accurately predict the biology of natural viruses. Finally, a Vero cell-adapted cDNA clone of ZIKV was generated that can be used as a convenient platform for studies aimed at the development of ZIKV vaccines and therapeutics.
  
  
  
  The availability of genetic tools and laboratory models determines the progress in understanding mechanisms of virus emergence and pathogenesis. Recent large-scale outbreaks of Zika virus (ZIKV) that were linked to complications during perinatal development and Guillain-Barré syndrome in adults emphasize the urgency for the development of a reverse-genetics system based on an epidemic ZIKV strain. Here, we report a stable infectious cDNA clone for ZIKV isolated during the 2015 epidemic in Brazil, as well as a Vero cell-adapted version of it, which will be used for virus-host interaction studies and vaccine development.
authors.affiliation: Laboratory of Infectious Diseases, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, Maryland, USA.
authors.forename: Konstantin A
authors.initials: KA
authors.lastname: Tsetsarkin
authors.affiliation: Laboratory of Infectious Diseases, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, Maryland, USA.
authors.forename: Heather
authors.initials: H
authors.lastname: Kenney
authors.affiliation: Department of Pathology, Institute for Human Infections and Immunity, University of Texas Medical Branch, Galveston, Texas, USA.
authors.forename: Rubing
authors.initials: R
authors.lastname: Chen
authors.affiliation: Laboratory of Infectious Diseases, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, Maryland, USA.
authors.forename: Guangping
authors.initials: G
authors.lastname: Liu
authors.affiliation: Center for Biologics Evaluation and Research, U.S. Food and Drug Administration, Silver Spring, Maryland, USA.
authors.forename: Hasmik
authors.initials: H
authors.lastname: Manukyan
authors.affiliation: Laboratory of Infectious Diseases, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, Maryland, USA.
authors.forename: Stephen S
authors.initials: SS
authors.lastname: Whitehead
authors.affiliation: Center for Biologics Evaluation and Research, U.S. Food and Drug Administration, Silver Spring, Maryland, USA.
authors.forename: Majid
authors.initials: M
authors.lastname: Laassri
authors.affiliation: Center for Biologics Evaluation and Research, U.S. Food and Drug Administration, Silver Spring, Maryland, USA.
authors.forename: Konstantin
authors.identifier: 0000-0003-3002-6247
authors.initials: K
authors.lastname: Chumakov
authors.affiliation: Laboratory of Infectious Diseases, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, Maryland, USA apletnev@niaid.nih.gov.
authors.forename: Alexander G
authors.initials: AG
authors.lastname: Pletnev
mesh_terms: D001938:Brazil / Q000453:epidemiology; D002460:Cell Line; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000235:genetics; D004279:DNA, Viral / Q000235:genetics; D004196:Disease Outbreaks; D054884:Host-Pathogen Interactions; D006801:Humans; D059386:Reverse Genetics / Q000379:methods*; D014765:Viral Vaccines / Q000276:immunology / Q000302:isolation & purification; D014773:Virology / Q000379:methods*; D014776:Virus Cultivation; D000071244:Zika Virus / Q000235:genetics* / Q000254:growth & development* / Q000302:isolation & purification; D000071243:Zika Virus Infection / Q000453:epidemiology / Q000821:virology*
chemical_list: D018076:DNA, Complementary; D004279:DNA, Viral; D014765:Viral Vaccines