Viruses

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    Protocols in Current Issue
    In vitro Membrane Interaction and Liposome Fusion Assays Using Recombinant Hepatitis C Virus Envelope Protein E2
    [Abstract] In order to study the mechanism underlying the Hepatitis C Virus (HCV) fusion process we have performed assays using phospholipid liposomes and a truncated form of E2 protein, E2661 (amino acids 384-661 of the HCV polyprotein) lacking the transmembrane region. E2661 has been previously generated by using the baculovirus ...
    Detection and Differentiation of Multiple Viral RNAs Using Branched DNA FISH Coupled to Confocal Microscopy and Flow Cytometry
    Authors:  Nicholas van Buuren and Karla Kirkegaard, date: 10/20/2018, view: 1621, Q&A: 0
    [Abstract] Due to the exceptionally high mutation rates of RNA-dependent RNA polymerases, infectious RNA viruses generate extensive sequence diversity, leading to some of the lowest barriers to the development of antiviral drug resistance in the microbial world. We have previously discovered that higher barriers to the development of drug resistance can be ...
    HCV Reporter System (Viral Infection-Activated Split-Intein-Mediated Reporter System) for Testing Virus Cell-to-cell Transmission ex-vivo
    Authors:  Fanfan Zhao, Ting Zhao, Libin Deng, Dawei Lv, Xiaolong Zhang, Xiaoyu Pan, Jun Xu and Gang Long, date: 08/05/2018, view: 1648, Q&A: 0
    [Abstract] Hepatitis C virus (HCV) spread involves two distinct entry pathways: cell-free transmission and cell-to-cell transmission. Cell-to-cell transmission is not only an efficient way for viruses to spread but also an effective method for escaping neutralizing antibodies. We adapted the viral infection-activated split-intein-mediated reporter system ...
    Virucidal and Neutralizing Activity Tests for Antiviral Substances and Antibodies
    Authors:  Chie Aoki-Utsubo, Ming Chen and Hak Hotta, date: 05/20/2018, view: 2966, Q&A: 0
    [Abstract] In a narrow definition, virucidal activity represents the activity by which to interact with and physically disrupt viral particles. In a broad definition, it includes the activity by which to functionally inhibit (neutralize) viral infectivity without apparent morphological alterations of the viral particles. The viral infectivity can be measured ...
    Time-of-addition and Temperature-shift Assays to Determine Particular Step(s) in the Viral Life Cycle that is Blocked by Antiviral Substance(s)
    Authors:  Chie Aoki-Utsubo, Ming Chen and Hak Hotta, date: 05/05/2018, view: 2281, Q&A: 0
    [Abstract] Viruses infect their host cells to produce progeny virus particles through the sequential steps of the viral life cycle, such as viral attachment, entry, penetration and post-entry events. This protocol describes time-of-addition and temperature-shift assays that are employed to explore which step(s) in the viral life cycle is blocked by an ...
    Isolation, BODIPY Labeling and Uptake of Exosomes in Hepatic Stellate Cells
    Author:  Shubham Shrivastava, date: 12/05/2017, view: 4414, Q&A: 1
    [Abstract] Exosomes have emerged as an important mediator of intercellular communication. They are present in extracellular milieu and therefore, easily accessible by neighboring or distant cells. They carry mRNA, microRNAs and proteins within their vesicles and once internalized by recipient cells; they can modulate multiple signaling pathways with ...
    Ultradeep Pyrosequencing of Hepatitis C Virus to Define Evolutionary Phenotypes
    [Abstract] Analysis of hypervariable regions (HVR) using pyrosequencing techniques is hampered by the ability of error correction algorithms to account for the heterogeneity of the variants present. Analysis of between-sample fluctuations to virome sub-populations, and detection of low frequency variants, are unreliable through the application of arbitrary ...
    Membrane Flotation Assay
    Authors:  Dorothee A Vogt and Melanie Ott, date: 04/05/2015, view: 8821, Q&A: 0
    [Abstract] Many postitive-stranded RNA viruses, such as Hepatitis C virus (HCV), highjack cellular membranes, including the Golgi, ER, mitchondria, lipid droplets, and utilize them for replication of their RNA genome or assembly of new virions. By investigating how viral proteins associate with cellular membranes we will better understand the roles of ...
    Infection Experiments (Hepatitis C Virus)
    Authors:  Lidia Mingorance, Claudia Vasallo, Martina Friesland and Pablo Gastaminza, date: 02/05/2015, view: 6058, Q&A: 0
    [Abstract] The establishment of a cell culture system for hepatitis C virus based on the JFH-1 strain and human hepatoma cell lines has been instrumental for the study of the viral replication cycle. The robustness of the JFH1-based cell culture models enabled many laboratories around the world to perform HCV infections in cell culture, accelerating the ...
    Hepatitis C virus Cell-to-cell Spread Assay
    Authors:  Naina Barretto and Susan L. Uprichard, date: 12/20/2014, view: 6765, Q&A: 1
    [Abstract] Hepatitis C virus (HCV) can infect naïve cells via entry of “cell-free” extracellular virus or direct “cell-to-cell” transmission. Here, we describe an assay for detecting HCV cell-to-cell transmission, using a non-growing cell culture system that avoids confounding effects of cell growth. The assay consists of infecting a small number of cells in ...



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