Abstract
Varicella zoster virus (VZV) is a human herpesvirus which causes Varicella (chickenpox) upon primary infection and Zoster (shingles) following reactivation from latency (von Bokay, 1909). Whilst VZV is extensively studied, inherent features of VZV replication, such as cell-association of virus particles during in vitro culture and a restricted host range (limited to humans and some other primates) mean the cellular and viral mechanisms underlying VZV reactivation and pathogenesis remain largely uncharacterised. Much remains to be learnt about VZV, interactions with its host, and the development of disease. This protocol describes a basic VZV replication assay using a recombinant VZV-GFP reporter virus. As VZV is highly cell-associated in tissue culture, the reporter virus inoculum described here is a preparation of infected cells. This reporter virus-infected cell line can be used in combination with siRNA gene depletion or cDNA overexpression transfection protocols to determine the effect of individual cellular genes on virus replication.
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Acknowledgments
We are grateful to Prof. Ann Arvin (Stanford School of Medicine) for the kind gift of the VZV-GFP clone, and to Dr. Armin Baiker for the propagation of a cell-associated VZV-GFP virus stock. The protocol was optimised by Dr Lakshmi N. Kaza and Dr Samantha Griffiths. The authors gratefully acknowledge the MRC for funding (G0501453 J.H.).
References
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