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Live Cell Imaging of Hepatitis C Virus Trafficking in Hepatocytes

  • Yasmine Baktash
  • Glenn Randall
Protocol
Part of the Methods in Molecular Biology book series (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.

Key words

Hepatitis C virus Fluorescent labeling Live cell imaging Viral entry DiD Tetracysteine tag TC-core 

Notes

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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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  1. 1.Department of MicrobiologyThe University of ChicagoChicagoUSA

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