Protocols in Current Issue
    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: 4575, 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: 10752, 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: 7065, 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: 9772, 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 ...
    Membrane Flotation Assay
    Authors:  Dorothee A Vogt and Melanie Ott, date: 04/05/2015, view: 12424, 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: 8448, 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: 9923, 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 ...
    Purification of HCV-remodeled and Control ER Membranes
    Authors:  David Paul and Ralf Bartenschlager, date: 05/20/2014, view: 7277, Q&A: 0
    [Abstract] As for all positive strand RNA viruses, hepatitis C virus (HCV) RNA replication is tightly associated with rearranged host cell membranes, termed viral replication factories. However, up to now little is known about both viral and cellular constituents of viral replication factories. Here, we describe a protocol to specifically isolate ...
    Colony Forming Assay for HCV-Replicon Cell Line
    Authors:  Chang Ho Lee and Seong-Wook Lee, date: 12/20/2013, view: 8931, Q&A: 0
    [Abstract] Hepatitis C virus (HCV) is the main causative agent of chronic hepatitis, liver cirrhosis, and hepatocellular carcinoma. Since the HCV genome is present exclusively in RNA form during replication, a number of anti-HCV drugs show appearance of rapid drug-resistant viruses. Therefore, it is important to test generation of drug-escape mutant viruses ...

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