title: Development and Characterization of Recombinant Virus Generated from a New World Zika Virus Infectious Clone.
abstract: Zika virus (ZIKV; family Flaviviridae, genus Flavivirus) is a rapidly expanding global pathogen that has been associated with severe clinical manifestations, including devastating neurological disease in infants. There are currently no molecular clones of a New World ZIKV available that lack significant attenuation, hindering progress toward understanding determinants of transmission and pathogenesis. Here we report the development and characterization of a novel ZIKV reverse genetics system based on a 2015 isolate from Puerto Rico (PRVABC59). We generated a two-plasmid infectious clone system from which infectious virus was rescued that replicates in human and mosquito cells with growth kinetics representative of wild-type ZIKV. Infectious clone-derived virus initiated infection and transmission rates in Aedes aegypti mosquitoes comparable to those of the primary isolate and displayed similar pathogenesis in AG129 mice. This infectious clone system provides a valuable resource to the research community to explore ZIKV molecular biology, vaccine development, antiviral development, diagnostics, vector competence, and disease pathogenesis.
  
  
  OBJECTIVE
  ZIKV is a rapidly spreading mosquito-borne pathogen that has been linked to Guillain-Barré syndrome in adults and congenital microcephaly in developing fetuses and infants. ZIKV can also be sexually transmitted. The viral molecular determinants of any of these phenotypes are not well understood. There is no reverse genetics system available for the current epidemic virus that will allow researchers to study ZIKV immunity, develop novel vaccines, or develop antiviral drugs. Here we provide a novel infectious clone system generated from a recent ZIKV isolated from a patient infected in Puerto Rico. This infectious clone produces virus with in vitro and in vivo characteristics similar to those of the primary isolate, providing a critical tool to study ZIKV infection and disease.
authors: Department of Microbiology, Immunology, and Pathology, Colorado State University, Fort Collins, Colorado, USA.
authors: James
authors: J
authors: Weger-Lucarelli
authors: Division of Vector-Borne Diseases, Centers for Disease Control and Prevention, Fort Collins, Colorado, USA.
authors: Nisha K
authors: NK
authors: Duggal
authors: Department of Microbiology, Immunology, and Pathology, Colorado State University, Fort Collins, Colorado, USA.
authors: Kristen
authors: K
authors: Bullard-Feibelman
authors: Department of Microbiology, Immunology, and Pathology, Colorado State University, Fort Collins, Colorado, USA.
authors: Milena
authors: M
authors: Veselinovic
authors: Department of Microbiology, Immunology, and Pathology, Colorado State University, Fort Collins, Colorado, USA.|Division of Vector-Borne Diseases, Centers for Disease Control and Prevention, Fort Collins, Colorado, USA.
authors: Hannah
authors: H
authors: Romo
authors: Department of Microbiology, Immunology, and Pathology, Colorado State University, Fort Collins, Colorado, USA.
authors: Chilinh
authors: C
authors: Nguyen
authors: Department of Microbiology, Immunology, and Pathology, Colorado State University, Fort Collins, Colorado, USA.
authors: Claudia
authors: C
authors: Rückert
authors: Division of Vector-Borne Diseases, Centers for Disease Control and Prevention, Fort Collins, Colorado, USA.
authors: Aaron C
authors: AC
authors: Brault
authors: Department of Microbiology, Immunology, and Pathology, Colorado State University, Fort Collins, Colorado, USA.
authors: Richard A
authors: RA
authors: Bowen
authors: Department of Microbiology, Immunology, and Pathology, Colorado State University, Fort Collins, Colorado, USA.
authors: Mark
authors: M
authors: Stenglein
authors: Department of Microbiology, Immunology, and Pathology, Colorado State University, Fort Collins, Colorado, USA brian.geiss@colostate.edu gregory.ebel@colostate.edu.|School of Biomedical Engineering, Colorado State University, Fort Collins, Colorado, USA.
authors: Brian J
authors: BJ
authors: Geiss
authors: Department of Microbiology, Immunology, and Pathology, Colorado State University, Fort Collins, Colorado, USA brian.geiss@colostate.edu gregory.ebel@colostate.edu.
authors: Gregory D
authors: GD
authors: Ebel
mesh_terms: D000330:Aedes / Q000821:virology*; D000818:Animals; D045744:Cell Line, Tumor; D002522:Chlorocebus aethiops; D002999:Clone Cells; D003001:Cloning, Molecular; D012321:DNA-Directed RNA Polymerases / Q000235:genetics / Q000378:metabolism; D004847:Epithelial Cells / Q000821:virology; D005818:Genetic Engineering; D003698:Hepatitis Delta Virus / Q000737:chemistry; D022781:Hepatocytes / Q000821:virology; D006801:Humans; D007303:Insect Vectors / Q000821:virology*; D051379:Mice; D010957:Plasmids / Q000737:chemistry / Q000378:metabolism*; D016337:RNA, Catalytic / Q000235:genetics / Q000378:metabolism; D059386:Reverse Genetics / Q000379:methods*; D016019:Survival Analysis; D014709:Vero Cells; D019562:Viral Load; D014764:Viral Proteins / Q000235:genetics / Q000378:metabolism; D014779:Virus Replication; D000071244:Zika Virus / Q000235:genetics* / Q000254:growth & development; D000071243:Zika Virus Infection / Q000401:mortality / Q000821:virology*
chemical_list: D016337:RNA, Catalytic; D014764:Viral Proteins; C062210:bacteriophage T7 RNA polymerase; D012321:DNA-Directed RNA Polymerases
keywords: Flavivirus; Zika virus; infectious clones