A luciferase-based cell-cell fusion assay for measuring dengue and Zika virus infections Hsiao-Han Lin 1, Suh-Chin Wu 1,2 1 Institute of Biotechnology, National Tsing Hua University, Taiwan; 2 Department of Medical Science, National Tsing Hua University, Taiwan
The type II membrane fusion induced by flavivirus E proteins is an unique pH-dependent membrane fusion process differently from the typical type I memberne fusion triggered by HIV and influenza viruses. The fusion peptide on the DII of the flavivirus E proteins can insert into the cell membrane as a cell entry process besides the receptor bindings. Traditional assay using C6/36 mosquito cells infected by dengue viruses has been previously reported but did not provide the efficient quantitation to measure the virus-triggered membrane fusion. Here we reported the development of quantitative cell fusion assay for four serotypes of dengue viruses and the recently emerged Zika viruses. We used a pCI-neo vector encoding the the prM and E genes of dengue and Zika viruses and investigated the cell fusion in transfected 293, Vero and CHO cells. Target cells were co-transfection of the dengue and Zika prME genes and T7 RNA polymerase to react with the effector cells transfected with the luciferase gene under the control of the T7 promoter. Quantification of the virus-induced cell fusion was determined by the luciferase expression levels under a switch of pH from 7.4 to 5.4. The quantitative cell-based assays were used for measuring the anti-fusion properties by two monoclonal antibodies 4G2 and DB42 against dengue and Zika virus infections.
Credits: None available.
You must be logged in and own this product in order to post comments.