A convenient cell fusion assay for the study of SARS-CoV entry and inhibition.
SARS-CoV spike (S) protein-mediated cell fusion is important for the viral entry mechanism and identification of SARS-CoV entry inhibitors. In order to avoid the high risks involved in handling SARS-CoV and to facilitate the study of viral fusion mechanism, we established the cell lines: SR-COS7 cells that stably express both SARS-CoV S protein and red fluorescence protein, R-COS7 cells that stably express red fluorescence protein, and AG-COS7 cells that stably express both ACE2 and green fluorescence protein, respectively. When SR-COS7 cells or R-COS7 cells were cocultured with AG-COS7 cells, syncytia with yellow fluorescence were conveniently observed after 12 h in SR-COS7 cells plus AG-COS7 cells, but not in R-COS7 cells plus AG-COS7 cells. The cell-to-cell fusion efficiency was simply determined for quantitative analysis based on the number of syncytium detected by flow cytometry. Such new cell-to-cell fusion model was further assessed by the potent HR2 peptide inhibitor, which led to the obvious decrease of the cell-to-cell fusion efficiency. The successful fusion and inhibition of cell-based binding assay shows that it can be well used for the study of SARS-CoV entry and inhibition.
Sha
YongGang
Y
State Key Laboratory of Virology, College of Life Sciences, Wuhan University, Wuhan, PR China.
Wu
YingLiang
Y
Cao
ZhiJian
Z
Xu
XiuLing
X
Wu
WenLan
W
Jiang
DaHe
D
Mao
Xin
X
Liu
Hui
H
Zhu
Ying
Y
Gong
Rui
R
Li
WenXin
W
D000085962:Angiotensin-Converting Enzyme 2; D000818:Animals; D001681:Biological Assay; D019556:COS Cells; D002459:Cell Fusion; D002522:Chlorocebus aethiops; D018920:Coculture Techniques; D004926:Escherichia coli / Q000235:genetics; D005456:Fluorescent Dyes / Q000378:metabolism; D005982:Glutathione Transferase / Q000378:metabolism; D049452:Green Fluorescent Proteins / Q000378:metabolism; D008164:Luminescent Proteins / Q000378:metabolism; D008562:Membrane Glycoproteins / Q000235:genetics / Q000378:metabolism*; D010455:Peptides / Q000737:chemistry / Q000378:metabolism; D007703:Peptidyl-Dipeptidase A / Q000378:metabolism; D011994:Recombinant Proteins / Q000378:metabolism; D045473:Severe acute respiratory syndrome-related coronavirus / Q000235:genetics / Q000378:metabolism*; D045169:Severe Acute Respiratory Syndrome / Q000378:metabolism*; D064370:Spike Glycoprotein, Coronavirus; D013997:Time Factors; D014357:Trypsin / Q000494:pharmacology; D014759:Viral Envelope Proteins / Q000235:genetics / Q000378:metabolism*; D000097573:Red Fluorescent Protein
D000085962:Angiotensin-Converting Enzyme 2; D005456:Fluorescent Dyes; D005982:Glutathione Transferase; D049452:Green Fluorescent Proteins; D008164:Luminescent Proteins; D008562:Membrane Glycoproteins; D010455:Peptides; D007703:Peptidyl-Dipeptidase A; D011994:Recombinant Proteins; D064370:Spike Glycoprotein, Coronavirus; D014357:Trypsin; D014759:Viral Envelope Proteins; D000097573:Red Fluorescent Protein; C578557:spike glycoprotein, SARS-CoV; C578553:spike protein, mouse hepatitis virus