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Updated: Jan 11, 2026
05:21
A Luciferase-fluorescent Reporter Influenza Virus for Live Imaging and Quantification of Viral Infection
Published on: August 14, 2019
26.0K
Tandem split-GFP influenza A viruses for sensitive and accurate replication analyses.
Himadri Nath1, Ariel Arndt1, Joshua T Mann1
1Infectious Disease Program, Lovelace Biomedical Research Institute, Albuquerque, New Mexico, USA.
Microbiology Spectrum
|November 17, 2025
Summary
Researchers developed a new influenza A virus (IAV) reporter system using split-green fluorescent protein (GFP) for stable and sensitive tracking of infections. This tool accelerates antiviral drug screening and host-virus interaction studies.
Area of Science:
- Virology
- Molecular Biology
- Biotechnology
Background:
- Influenza A viruses (IAVs) are crucial for studying viral fitness, antiviral efficacy, and host interactions.
- Engineering stable IAV reporter viruses is challenging due to the virus's compact and segmented genome.
- Existing reporter systems can compromise viral function or stability.
Purpose of the Study:
- To develop a genetically stable, replication-competent IAV reporter virus with minimal fitness defects.
- To create a sensitive and scalable system for monitoring IAV replication and facilitating high-throughput screening.
- To enable comparative studies of IAV host-virus interactions across different species.
Main Methods:
- Developed a replication-competent IAV incorporating a tandem split-GFP reporter strategy (GFP11×7) fused to polymerase subunits (PB2 or PA).
- Utilized a self-cleaving 2A peptide for either polymerase fusion proteins or released GFP.
- Employed *trans*-complementation with cells expressing GFP1-10 for bright fluorescence.
- Engineered GFP1-10-expressing cell lines from multiple vertebrate species.
Main Results:
- The tandem GFP11 reporter viruses demonstrated genetic stability over serial passages with strong correlation between GFP signal and viral gene expression.
- Viruses exhibited delayed replication kinetics but maintained high fluorescence.
- The system proved effective for high-throughput quantification of infection, neutralizing antibody titration, antiviral drug screening, and host factor identification.
- Results from the reporter system closely matched traditional assays but with accelerated timelines.
- Broad compatibility was shown across engineered cell lines from various vertebrate species.
Conclusions:
- The GFP11×7 reporter platform provides a versatile, robust, and sensitive tool for influenza A virus research and therapeutic screening.
- This system enables scalable and quantitative evaluation of IAV replication in diverse experimental contexts.
- The platform facilitates rapid adaptation for emerging IAV strains, supporting pandemic preparedness and antiviral development.
