PMID- 34806704
OWN - NLM
STAT- MEDLINE
DCOM- 20211125
LR  - 20230620
IS  - 1940-087X (Electronic)
IS  - 1940-087X (Linking)
IP  - 177
DP  - 2021 Nov 5
TI  - Live Imaging and Quantification of Viral Infection in K18 hACE2 Transgenic Mice 
      Using Reporter-Expressing Recombinant SARS-CoV-2.
LID - 10.3791/63127 [doi]
AB  - 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.
FAU - Morales Vasquez, Desarey
AU  - Morales Vasquez D
AD  - Texas Biomedical Research Institute.
FAU - Chiem, Kevin
AU  - Chiem K
AD  - Texas Biomedical Research Institute.
FAU - Silvas, Jesus
AU  - Silvas J
AD  - Texas Biomedical Research Institute.
FAU - Park, Jun-Gyu
AU  - Park JG
AD  - Texas Biomedical Research Institute.
FAU - Ye, Chengjin
AU  - Ye C
AD  - Texas Biomedical Research Institute; CYe@txbiomed.org.
FAU - Martinez-Sobrido, Luis
AU  - Martinez-Sobrido L
AD  - Texas Biomedical Research Institute; LMartinez@txbiomed.org.
LA  - eng
GR  - 75N93021C00014/AI/NIAID NIH HHS/United States
GR  - R01 AI161175/AI/NIAID NIH HHS/United States
GR  - R01 AI161363/AI/NIAID NIH HHS/United States
GR  - R43 AI165089/AI/NIAID NIH HHS/United States
PT  - Journal Article
PT  - Video-Audio Media
DEP - 20211105
PL  - United States
TA  - J Vis Exp
JT  - Journal of visualized experiments : JoVE
JID - 101313252
RN  - 0 (KRT18 protein, human)
RN  - 0 (Keratin-18)
RN  - EC 3.4.17.23 (Angiotensin-Converting Enzyme 2)
SB  - IM
MH  - Angiotensin-Converting Enzyme 2/*genetics
MH  - Animals
MH  - *COVID-19
MH  - Humans
MH  - Keratin-18/*genetics
MH  - Mice
MH  - Mice, Transgenic
MH  - SARS-CoV-2
MH  - *Virus Diseases
PMC - PMC10276921
MID - NIHMS1901122
EDAT- 2021/11/23 06:00
MHDA- 2021/11/26 06:00
PMCR- 2023/06/18
CRDT- 2021/11/22 08:52
PHST- 2021/11/22 08:52 [entrez]
PHST- 2021/11/23 06:00 [pubmed]
PHST- 2021/11/26 06:00 [medline]
PHST- 2023/06/18 00:00 [pmc-release]
AID - 10.3791/63127 [doi]
PST - epublish
SO  - J Vis Exp. 2021 Nov 5;(177):10.3791/63127. doi: 10.3791/63127.