Classen, Nica; Ulrich, Diana; Hofemeier, Arne D.; Hennies, Marc Tim; Hafezi, Wali; Pettke, Aleksandra; Romberg, Marie-Luise; Lorentzen, Eva U.; Hensel, Andreas; Kühn, Joachim: Broadly Applicable, Virus-Free Dual Reporter Assay to Identify Compounds Interfering with Membrane Fusion: Performance for HSV-1 and SARS-CoV-2
Inhalt
- Materials and Methods
- Plasmids
- Cell Culture
- Stable Reporter Cell Lines
- Stable Reporter Fusion Assay (SRFIA) with HSV-1 Fusion Machinery
- Stable Reporter Fusion Assay (SRFIA) with SARS-CoV-2 Fusion Machinery
- Measurement of NLuc Activity
- Cytotoxicity Assay XTT
- Microscopical Determination of Syncytia Formation
- Calculation of Zhang Indices
- Statistics
- Results
- Bicistronic Reporter Gene Cassette and Stable Vero Reporter Cell Lines
- HSV-1 Fusion Machinery Induces Syncytia in Vero Reporter Cells
- HSV-1 Glycoprotein Mediated Cell–Cell Fusion Results in SEAP Reporter Gene Expression
- Effect of Known Inhibitors of HSV-1 Cell–Cell Fusion on Reporter Gene Expression
- Establishment of Dual secNL/SEAP SRFIA
- Performance of HSV-1-Specific SRFIA as Screening Assay for Fusion Inhibitors on 96-Well Plates
- SARS-CoV-2 S Protein Induces Formation of Syncytia and SEAP Expression in Reporter Cells
- Effect of Umifenovir on Syncytia Formation by the S Protein
- Performance of S Protein-Specific SRFIA as Screening Assay on 96-Well Plates
- Discussion
- References