Details

Autor(en) / Beteiligte

• Xu, Chang
• Ren, Xiao-He
• Han, Di
• Peng, Yan
• Lei, Jin-Ju
• Yu, Luo-Xiao
• Liu, Ling-Juan
• Xu, Wei-Chao
• Cheng, Si-Xue

Titel

Precise Detection on Cell–Cell Fusion by a Facile Molecular Beacon-Based Method

Ist Teil von

• Analytical chemistry (Washington), 2022-12, Vol.94 (49), p.17334-17340

Ort / Verlag

United States: American Chemical Society

Erscheinungsjahr

2022

Quelle

MEDLINE

Beschreibungen/Notizen

• Cell–cell fusion studies provide an experimental platform for evaluating disease progression and investigating cell infection. However, to realize sensitive and quantitative detection on cell–cell fusion is still a challenge. Herein, we report a facile molecular beacon (MB)-based method for precise detection on cell–cell fusion. By transfection of the spike protein (S protein) and enhanced green fluorescent protein (EGFP) in HEK 293 cells, the virus-mimicking fusogenic effector cells 293-S-EGFP cells were constructed to interact with target cells. Before mixing the effector cells with the target cells, the glyceraldehyde-3-phosphate dehydrogenase (GAPDH) expression in 293-S-EGFP cells was silenced, and the MB for GAPDH mRNA detection was delivered into the GAPDH silenced 293-S-EGFP cells. Once cell–cell fusion occurred, MB migrated from the GAPDH silenced effector cells to the target cells and hybridized with GAPDH mRNA in the target cells to induce fluorescence emission. The cell–cell fusion can be easily visualized and quantitated by fluorescence microscopy and flow cytometry. The fluorescence intensity is strongly dependent on the number of fused target cells. This MB-based method can easily identify the differences in the cell fusions for various target cells with different angiotensin-converting enzyme 2 (ACE2) and transmembrane serine protease 2 (TMPRSS2) expression levels, resulting in dramatically different fluorescence intensities in fused target cells. Our study provides a convenient and efficient quantitative detection approach to study cell–cell fusion.