Visualizing infection spread: dual-color fluorescent reporting of virus-host interactions

Biotechnology and Bioengineering
Adam SwickJohn Yin

Abstract

Although the molecular mechanisms by which host cells defend themselves against viral infection have been studied in great depth, and countermeasures viruses employ to suppress such defensive responses have been widely documented, relatively little attention has been devoted toward elucidating how such interactions between virus and host are resolved over multiple rounds of infection. Here, we describe the design, synthesis, and validation of a dual-color fluorescent reporter system to study how viral infections spread through a host cell monolayer and how the cellular innate immune system mounts an antiviral response. We employed recombinant, red fluorescent protein expressing mutants of a prototypical RNA virus, vesicular stomatitis virus to enable identification and tracking of infected cells. Further, we generated stable reporter cells that use green fluorescent protein to report on the expression of IFIT2, an interferon stimulated gene involved in the interference of viral protein translation, and a marker of antiviral defense activation. The presence of the fluorescent protein reporters had minimal effects on the normal behavior of the cells or viruses. Moreover, expression of the virus and cell reporters correlated with ...Continue Reading

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Citations

Feb 18, 2016·Analytical Chemistry·Gregory K PottsJoshua J Coon
Apr 18, 2015·Frontiers in Genetics·Kendra PeskoPaul E Turner
Sep 13, 2016·Annual Review of Virology·Frederik Graw, Alan S Perelson
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Aug 24, 2017·Integrative Biology : Quantitative Biosciences From Nano to Macro·Andrea C TimmJohn Yin

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