A Luciferase-Fluorescent Reporter Influenza Virus to Track Viral Infections

Abstract

Source: Chiem, K., et al. A Luciferase-fluorescent Reporter Influenza Virus for Live Imaging and Quantification of Viral Infection. J. Vis. Exp. (150), (2019).

This video demonstrates a method to track viral infections in mouse models using a recombinant luciferase and fluorescence protein-expressing bi-reporter Influenza A virus. This method enables both in vivo and ex vivo studies, enabling researchers to gain insights into viral dynamics and the effects of interventions.

Protocol

All procedures involving animal models have been reviewed by the local institutional animal care committee and the JoVE veterinary review board. 1. In Vivo Characterization of BIRFLU (Figure 1 and Figure 2) Mouse infection NOTE: Intranasal infection of mice was done as previously described. For a more detailed protocol of IAV infection in vivo using the mouse model of infec…

Representative Results

Figure 1: Schematic representation for the study of BIRFLU in mice. Expression of Nluc and Venus reporter genes was evaluated in mice infected with 1 x 106 PFU of BIRFLU using in vivo or ex vivo imaging. Briefly, on day 1, 5 to 7-week-old female BALB/c mice were mock-infected (1x PBS) or inoculated with 1 x 106 PFU of BIRFLU intranasally. At day 3 post-infe…

Materials

Five- to seven-week-old female BALB/c mice	National Cancer Institute (NCI)	555
Isoflurane	Baxter	1001936040	Store at Room temperature
IVIS Spectrum	PerkinElmer	124262	This instrument is used for in vivo imaging
IX81 Motorized Inverted Microscope	Olympus	Olympus IX81
Living Image 4.7.2 software	PerkinElmer		This instrument is used for in vivo imaging
Lumicount	Packard		This instrument is used for quantifying luciferase activity
Nano-Glo Luciferase Assay Reagent	Promega	N1110	This reagent is used to measure Nluc activity. Store at -20 °C
Retiga 20000R Fast1394 Camera	Qimaging	Retiga 2000R
Scanner	HP