title: An Improved Reverse Genetics System to Overcome Cell-Type-Dependent Ebola Virus Genome Plasticity.
abstract: Reverse genetics systems represent a key technique for studying replication and pathogenesis of viruses, including Ebola virus (EBOV). During the rescue of recombinant EBOV from Vero cells, a high frequency of mutations was observed throughout the genomes of rescued viruses, including at the RNA editing site of the glycoprotein gene. The influence that such genomic instability could have on downstream uses of rescued virus may be detrimental, and we therefore sought to improve the rescue system. Here we report an improved EBOV rescue system with higher efficiency and genome stability, using a modified full-length EBOV clone in Huh7 cells. Moreover, by evaluating a variety of cells lines, we revealed that EBOV genome instability is cell-type dependent, a fact that has significant implications for the preparation of standard virus stocks. Thus, our improved rescue system will have an impact on both basic and translational research in the filovirus field.
authors: Molecular Virology and Host-Pathogen Interaction Unit Department of Microbiology, Graduate School of Medicine, Hokkaido University, Sapporo, Japan.
authors: Yoshimi
authors: Y
authors: Tsuda
authors: Disease Modeling and Transmission Section, Laboratory of Virology.
authors: Thomas
authors: T
authors: Hoenen
authors: Molecular Virology and Host-Pathogen Interaction Unit.
authors: Logan
authors: L
authors: Banadyga
authors: Molecular Virology and Host-Pathogen Interaction Unit.
authors: Carla
authors: C
authors: Weisend
authors: Genomics Unit, Research Technology Section, Division of Intramural Research, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Hamilton, Montana.
authors: Stacy M
authors: SM
authors: Ricklefs
authors: Genomics Unit, Research Technology Section, Division of Intramural Research, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Hamilton, Montana.
authors: Stephen F
authors: SF
authors: Porcella
authors: Molecular Virology and Host-Pathogen Interaction Unit.
authors: Hideki
authors: H
authors: Ebihara
mesh_terms: D000818:Animals; D019556:COS Cells; D002460:Cell Line; D045744:Cell Line, Tumor; D002522:Chlorocebus aethiops; D029043:Ebolavirus / Q000235:genetics*; D016679:Genome, Viral / Q000235:genetics*; D042822:Genomic Instability / Q000235:genetics; D006023:Glycoproteins / Q000235:genetics; D057809:HEK293 Cells; D019142:Hemorrhagic Fever, Ebola / Q000821:virology*; D006801:Humans; D009154:Mutation / Q000235:genetics; D059386:Reverse Genetics / Q000379:methods; D014709:Vero Cells; D014779:Virus Replication / Q000235:genetics
chemical_list: D006023:Glycoproteins
keywords: Ebola virus; RNA editing site; mutation; reverse genetics system