Promoting Simultaneous Onset of Viral Gene Expression Among Cells Infected with Herpes Simplex Virus-1

Synchronous viral infection facilitates the study of viral gene expression, viral host interactions, and viral replication processes. However, the protocols for achieving synchronous infections were hardly ever tested in proper temporal resolution at the single-cell level. We set up a fluorescence-b...

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Bibliographic Details
Published in:	Frontiers in microbiology Vol. 8; p. 2152
Main Authors:	Ralph, Maya, Bednarchik, Marina, Tomer, Enosh, Rafael, Dor, Zargarian, Sefi, Gerlic, Motti, Kobiler, Oren
Format:	Journal Article
Language:	English
Published:	Switzerland Frontiers Media S.A 01-11-2017
Subjects:	biological noise herpesviruses Microbiology single cell temperature sensitive timing of infection herpesviruses single cell timing of infection biological noise temperature sensitive
Online Access:	Get full text
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Summary:	Synchronous viral infection facilitates the study of viral gene expression, viral host interactions, and viral replication processes. However, the protocols for achieving synchronous infections were hardly ever tested in proper temporal resolution at the single-cell level. We set up a fluorescence-based, time lapse microscopy assay to study sources of variability in the timing of gene expression during herpes simplex virus-1 (HSV-1) infection. We found that with the common protocol, the onset of gene expression within different cells can vary by more than 3 h. We showed that simultaneous viral genome entry to the nucleus can be achieved with a derivative of the previously characterized temperature sensitive mutant tsB7, however, this did not improve gene expression synchrony. We found that elevating the temperature in which the infection is done and increasing the multiplicity of infection (MOI) significantly promoted simultaneous onset of viral gene expression among infected cells. Further, elevated temperature result in a decrease in the coefficient of variation (a standardized measure of dispersion) of viral replication compartments (RCs) sizes among cells as well as a slight increment of viral late gene expression synchrony. We conclude that simultaneous viral gene expression can be improved by simple modifications to the infection process and may reduce the effect of single-cell variability on population-based assays.
Bibliography:	ObjectType-Article-1 SourceType-Scholarly Journals-1 ObjectType-Feature-2 content type line 23 This article was submitted to Virology, a section of the journal Frontiers in Microbiology Edited by: Michael Nevels, University of St. Andrews, United Kingdom Reviewed by: Maria Kalamvoki, University of Kansas, United States; Lüder Wiebusch, Charité Universitätsmedizin Berlin, Germany
ISSN:	1664-302X 1664-302X
DOI:	10.3389/fmicb.2017.02152