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

Abstract

The coronavirus disease 2019 (COVID-19) pandemic has been caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). To date, SARS-CoV-2 has been responsible for over 242 million infections and more than 4.9 million deaths worldwide. Similar to other viruses, studying SARS-CoV-2 requires the use of experimental methods to detect the presence of virus in infected cells and/or in animal models. To overcome this limitation, we generated replication-competent recombinant (r)SARS-CoV-2 that expresses bioluminescent (nanoluciferase, Nluc) or fluorescent (Venus) proteins. These reporter-expressing rSARS-CoV-2 allow tracking viral infections in vitro and in vivo based on the expression of Nluc and Venus reporter genes. Here the study describes the use of rSARS-CoV-2/Nluc and rSARS-CoV-2/Venus to detect and track SARS-CoV-2 infection in the previously described K18 human angiotensin-converting enzyme 2 (hACE2) transgenic mouse model of infection using in vivo imaging systems (IVIS). This rSARS-CoV-2/Nluc and rSARS-CoV-2/Venus show rSARS-CoV-2/WT-like pathogenicity and viral replication in vivo. Importantly, Nluc and Venus expression allow us to directly track viral infections in vivo and ex vivo, in infected mice. These rSARS-CoV-2/Nluc and rSARS-CoV-2/Venus represent an excellent option to study the biology of SARS-CoV-2 in vivo, to understand viral infection and associated COVID-19 disease, and to identify effective prophylactic and/or therapeutic treatments to combat SARS-CoV-2 infection.

• Authors: Desarey Morales Vasquez, Kevin Chiem, Jesus Silvas, Jun-Gyu Park, Chengjin Ye, Luis Martínez-Sobrido
• Journal: Journal of visualized experiments : JoVE (J Vis Exp) Issue 177 (11 05 2021) ISSN: 1940-087X [Electronic] United States
• PMID: 34806704 (Publication Type: Journal Article, Video-Audio Media)

Chemical References

• KRT18 protein, human
• Keratin-18
• Angiotensin-Converting Enzyme 2

Topics

• Angiotensin-Converting Enzyme 2 (genetics)
• Animals
• COVID-19
• Humans
• Keratin-18 (genetics)
• Mice
• Mice, Transgenic
• SARS-CoV-2
• Virus Diseases