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Hepatitis C Virus Protocols pp 263–274Cite as

Live Cell Imaging of Hepatitis C Virus Trafficking in Hepatocytes

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Part of the book series: Methods in Molecular Biology ((MIMB,volume 1911))

Abstract

Standard fixed cell confocal microscopy is inherently limited in visualizing dynamic processes involving two- and three-dimensional movement. To overcome these limitations, live cell imaging approaches have been developed to study hepatitis C virus (HCV) entry, replicase protein trafficking, virion assembly, and egress. These studies have relied on fluorescent labeling of viral proteins by epitope tag insertion, genome labeling via nucleophilic dyes, or using lipophilic dyes to label the virion envelope. In this method review, we describe two approaches to study HCV virion trafficking in live cells. Lipophilic labeling of the envelope allows for study of the early events (through virion uncoating/fusion) in the HCV lifecycle. Tetracysteine (TC) tag insertion into the capsid protein permits study of virion assembly and capsid trafficking via binding of a fluorogenic biarsenical dye.

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Acknowledgments

We thank Kelly Coller Metzinger, The University of Chicago Light Microscopy Facility and its director Vytas Bindokas for helping develop this protocol. This work was funded by NIAID (AI080703). Y.B. was funded by NIH training grant T32 GM007183.

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Authors and Affiliations

  1. Department of Microbiology, The University of Chicago, Chicago, IL, USA

    Yasmine Baktash & Glenn Randall

Authors
  1. Yasmine Baktash
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  2. Glenn Randall
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Corresponding author

Correspondence to Glenn Randall .

Editor information

Editors and Affiliations

  1. Department of Immunology and Microbiology, The Scripps Research Institute, La Jolla, CA, USA

    Mansun Law

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Baktash, Y., Randall, G. (2019). Live Cell Imaging of Hepatitis C Virus Trafficking in Hepatocytes. In: Law, M. (eds) Hepatitis C Virus Protocols . Methods in Molecular Biology, vol 1911. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-8976-8_18

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  • DOI: https://doi.org/10.1007/978-1-4939-8976-8_18

  • Published:

  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-8975-1

  • Online ISBN: 978-1-4939-8976-8

  • eBook Packages: Springer Protocols

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