Optimisations and Challenges Involved in the Creation of Various Bioluminescent and Fluorescent Influenza A Virus Strains for In Vitro and In Vivo Applications

Spronken, Monique I., Short, Kirsty R., Herfst, Sander, Bestebroer, Theo M., Vaes, Vincent P., van der Hoeven, Barbara, Koster, Abraham J., Kremers, Gert-Jan, Scott, Dana P., Gultyaev, Alexander P., Sorell, Erin M., de Graaf, Miranda, Barcena, Montserrat, Rimmelzwaan, Guus F. and Fouchier, Ron A. (2015) Optimisations and Challenges Involved in the Creation of Various Bioluminescent and Fluorescent Influenza A Virus Strains for In Vitro and In Vivo Applications. PLoS One, 10 8: e0133888-e0133888. doi:10.1371/journal.pone.0133888


Author Spronken, Monique I.
Short, Kirsty R.
Herfst, Sander
Bestebroer, Theo M.
Vaes, Vincent P.
van der Hoeven, Barbara
Koster, Abraham J.
Kremers, Gert-Jan
Scott, Dana P.
Gultyaev, Alexander P.
Sorell, Erin M.
de Graaf, Miranda
Barcena, Montserrat
Rimmelzwaan, Guus F.
Fouchier, Ron A.
Title Optimisations and Challenges Involved in the Creation of Various Bioluminescent and Fluorescent Influenza A Virus Strains for In Vitro and In Vivo Applications
Journal name PLoS One   Check publisher's open access policy
ISSN 1932-6203
Publication date 2015-08-01
Year available 2015
Sub-type Article (original research)
DOI 10.1371/journal.pone.0133888
Open Access Status DOI
Volume 10
Issue 8
Start page e0133888
End page e0133888
Total pages 30
Place of publication San Francisco, CA United States
Publisher Public Library of Science
Language eng
Formatted abstract
Bioluminescent and fluorescent influenza A viruses offer new opportunities to study influenza virus replication, tropism and pathogenesis. To date, several influenza A reporter viruses have been described. These strategies typically focused on a single reporter gene (either bioluminescent or fluorescent) in a single virus backbone. However, whilst bioluminescence is suited to in vivo imaging, fluorescent viruses are more appropriate for microscopy. Therefore, the idea l reporter virus varies depending on the experiment in question, and it is important that any reporter virus strategy can be adapted accordingly. Herein, a strategy was developed to create five different reporter viruses in a single virus backbone. Specifically, enhanced green fluorescent protein (eGFP), far-red fluorescent protein (fRFP), near-infrared fluorescent protein (iRFP), Gaussia luciferase (gLUC) and firefly luciferase (fLUC) were inserted into the PA gene segment of A/PR/8/34 (H1N1). This study provides a comprehensive characterisation of the effects of different reporter genes on influenza virus replication and reporter activity. In vivo reporter gene expression, in lung tissues, was only detected for eGFP, fRFP and gLUC expressing viruses. In vitro, the eGFP-expressing virus displayed the best reporter stability and could be used for correlative light electron microscopy (CLEM). This strategy was then used to create eGFP-expressing viruses consisting entirely of pandemic H1N1, highly pathogenic avian influenza (HPAI) H5N1 and H7N9. The HPAI H5N1 eGFP-expressing virus infected mice and reporter gene expression was detected, in lung tissues, in vivo. Thus, this study provides new tools and insights for the creation of bioluminescent and fluorescent influenza A reporter viruses.
Keyword Neutralizing Antibodies
Electron Microscopy
Packaging Signals
Reporter Virus
Infection
Generation
Q-Index Code C1
Q-Index Status Confirmed Code
Institutional Status UQ

Document type: Journal Article
Sub-type: Article (original research)
Collections: Official 2016 Collection
School of Biomedical Sciences Publications
 
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