Live Imaging and Quantification of Viral Infection in K18 hACE2 Transgenic Mice Using Reporter-Expressing Recombinant SARS-CoV-2

Overview

This protocol describes the use of fluorescent or luminescent-expressing recombinant SARS-CoV-2 for in vivo and ex vivo live imaging of viral detection and tracking in K18 human ACE2 transgenic mice. This technique allows for effective evaluation of therapeutics for SARS-CoV-2 infection.

Key Study Components

Area of Science

• Virology
• In vivo imaging
• Transgenic mouse models

Background

• Understanding SARS-CoV-2 infection dynamics is crucial for therapeutic development.
• In vivo imaging systems (IVIS) facilitate real-time monitoring of viral behavior.
• K18 hACE2 transgenic mice are a relevant model for studying SARS-CoV-2.
• Fluorescent and luminescent reporters enhance detection capabilities.

Purpose of Study

• To develop a protocol for tracking SARS-CoV-2 infections in vivo.
• To assess pathogenicity, viral replication, and trophism using imaging techniques.
• To improve therapeutic evaluation methods for SARS-CoV-2.

Methods Used

• Use of luciferase- and fluorescence-expressing recombinant SARS-CoV-2.
• In vivo imaging with IVIS after substrate injection.
• Monitoring bioluminescence signal post-imaging setup.
• Shaving mice to enhance imaging clarity.

Main Results

• Effective tracking of SARS-CoV-2 in the K18 hACE2 transgenic mouse model.
• Demonstrated feasibility of in vivo and ex vivo imaging for viral studies.
• Provided insights into viral dynamics and therapeutic responses.
• Established a reliable method for assessing SARS-CoV-2 pathogenicity.

Conclusions

• The protocol offers a robust approach for studying SARS-CoV-2 infections.
• In vivo imaging enhances understanding of viral behavior and treatment efficacy.
• This technique can be adapted for other viral studies in transgenic models.

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