A Luciferase-Fluorescent Reporter Influenza Virus to Track Viral Infections
Published: January 31, 2024
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…
Declarações
The authors have nothing to disclose.
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 |
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A Luciferase-Fluorescent Reporter Influenza Virus to Track Viral Infections. J. Vis. Exp. (Pending Publication), e21896, doi: (2024).