| A Cre-based double fluorescence indicator system for monitoring cell fusion events and selection of fused cells. | |
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MedLine Citation:
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PMID: 20359294 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
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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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Authors:
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Kurt Pfannkuche; Dimitry Spitkovsky; Frank Thomas Wunderlich; Osama Mohamed Abd El Aziz; Tomo Saric; J?rgen Hescheler; Agapios Sachinidis |
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Publication Detail:
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Type: Journal Article; Research Support, Non-U.S. Gov't |
Journal Detail:
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Title: BioTechniques Volume: 48 ISSN: 1940-9818 ISO Abbreviation: BioTechniques Publication Date: 2010 Feb |
Date Detail:
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Created Date: 2010-04-02 Completed Date: 2010-06-28 Revised Date: - |
Medline Journal Info:
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Nlm Unique ID: 8306785 Medline TA: Biotechniques Country: England |
Other Details:
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Languages: eng Pagination: 113-20 Citation Subset: IM |
Affiliation:
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Institute for Neurophysiology, University of Cologne, Cologne, Germany. akp59@uni-koeln.de |
Export Citation:
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APA/MLA Format Download EndNote Download BibTex |
| MeSH Terms | |
Descriptor/Qualifier:
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Animals Cell Fusion / methods* Cell Line Genetic Vectors Humans Immunohistochemistry Integrases / genetics, metabolism* Luminescent Proteins / genetics, metabolism Mice Mice, Transgenic Microscopy, Fluorescence / methods* Molecular Biology / methods* Puromycin Receptors, Estrogen Reproducibility of Results |
| Chemical | |
Reg. No./Substance:
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0/Luminescent Proteins; 0/Receptors, Estrogen; 0/red fluorescent protein; 53-79-2/Puromycin; EC 2.7.7.-/Cre recombinase; EC 2.7.7.-/Integrases |
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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