title: A plasmid-based reporter system for live cell imaging of dengue virus infected cells.
abstract: Cell culture models are used widely to study the effects of dengue virus (DENV) on host cell function. Current methods of identification of cells infected with an unmodified DENV requires fixation and permeablization of cells to allow DENV-specific antibody staining. This method does not permit imaging of viable cells over time. In this report, a plasmid-based reporter was developed to allow non-destructive identification of DENV-infected cells. The plasmid-based reporter was demonstrated to be broadly applicable to the four DENV serotypes, including low-passaged strains, and was specifically cleaved by the viral protease with minimal interference on viral production. This study reveals the potential for this novel reporter system to advance the studies of virus-host interactions during DENV infection.
authors.affiliation: Institute for Immunology and Informatics, Department of Cell and Molecular Biology, University of Rhode Island, Providence, RI 02903, United States. Electronic address: cmedin.uri@gmail.com.
authors.forename: Carey L
authors.initials: CL
authors.lastname: Medin
authors.affiliation: Institute for Immunology and Informatics, Department of Cell and Molecular Biology, University of Rhode Island, Providence, RI 02903, United States.
authors.forename: Sierra
authors.initials: S
authors.lastname: Valois
authors.affiliation: Division of Infectious Diseases and Immunology, Department of Medicine, University of Massachusetts Medical School, Worcester, MA 01655, United States.
authors.forename: Chinmay G
authors.initials: CG
authors.lastname: Patkar
authors.affiliation: Institute for Immunology and Informatics, Department of Cell and Molecular Biology, University of Rhode Island, Providence, RI 02903, United States; Division of Infectious Diseases and Immunology, Department of Medicine, University of Massachusetts Medical School, Worcester, MA 01655, United States.
authors.forename: Alan L
authors.initials: AL
authors.lastname: Rothman
mesh_terms: D000818:Animals; D003584:Cytological Techniques / Q000379:methods*; D003716:Dengue Virus / Q000254:growth & development*; D017930:Genes, Reporter; D006801:Humans; D008856:Microscopy, Fluorescence / Q000379:methods*; D061848:Optical Imaging / Q000379:methods*; D010957:Plasmids; D014773:Virology / Q000379:methods
keywords: Dengue virus; Fluorescence microscopy; Protease; Viral reporter