A Cre-based double fluorescence indicator system for monitoring cell fusion events and selection of fused cells

A Cre-based double fluorescence indicator system for monitoring cell fusion events and selection of fused cells

We have established an in vitro Cre/loxP-based assay for monitoring cell fusion events that specifically traces the transport of cytoplasm from one cell to its fusion partner. Cells with a double fluorescence vector indicate fusion with cells expressing Cre recombinase by switching expression from r...

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Veröffentlicht in:BioTechniques. - 1993. - 48(2010), 2 vom: 01. Feb., Seite 113-20
1. Verfasser: Pfannkuche, Kurt (VerfasserIn)
Weitere Verfasser: Spitkovsky, Dimitry, Wunderlich, Frank Thomas, Abd El Aziz, Osama Mohamed, Saric, Tomo, Hescheler, Jürgen, Sachinidis, Agapios
Format: Online-Aufsatz
Sprache:English
Veröffentlicht: 2010
Zugriff auf das übergeordnete Werk:BioTechniques
Schlagworte:Journal Article Research Support, Non-U.S. Gov't Luminescent Proteins Receptors, Estrogen Puromycin 4A6ZS6Q2CL Cre recombinase EC 2.7.7.- Integrases
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520 |a We have established an in vitro Cre/loxP-based assay for monitoring cell fusion events that specifically traces the transport of cytoplasm from one cell to its fusion partner. Cells with a double fluorescence vector indicate fusion with cells expressing Cre recombinase by switching expression from red to green fluorescent protein through a Cre-mediated recombination event that simultaneously activates puromycin-acetyltransferase expression. This strategy allows for both the observation and puromycin selection of indicator cells that have undergone fusion with a Cre recombinase-expressing partner. A fusion protein of Cre with estrogen receptor (ER) can be used to control Cre recombinase activity through the tamoxifen-induced translocation of the Cre-ER fusion protein to the nucleus. Here we have established a new methodology that not only allows the monitoring of the transport of cellular contents, but also enables the purification of fused cells using puromycin 
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650 4 |a Research Support, Non-U.S. Gov't 
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700 1 |a Spitkovsky, Dimitry  |e verfasserin  |4 aut 
700 1 |a Wunderlich, Frank Thomas  |e verfasserin  |4 aut 
700 1 |a Abd El Aziz, Osama Mohamed  |e verfasserin  |4 aut 
700 1 |a Saric, Tomo  |e verfasserin  |4 aut 
700 1 |a Hescheler, Jürgen  |e verfasserin  |4 aut 
700 1 |a Sachinidis, Agapios  |e verfasserin  |4 aut 
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