Document Detail


A Cre-based double fluorescence indicator system for monitoring cell fusion events and selection of fused cells.
MedLine Citation:
PMID:  20359294     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
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.
Authors:
Kurt Pfannkuche; Dimitry Spitkovsky; Frank Thomas Wunderlich; Osama Mohamed Abd El Aziz; Tomo Saric; J?rgen Hescheler; Agapios Sachinidis
Related Documents :
19931884 - Different activities of the reovirus fast proteins and influenza hemagglutinin in cell-...
1890304 - The generation of immunogenic peptides can be selectively increased or decreased by pro...
12235284 - Domains of type 1 protein phosphatase inhibitor-2 required for nuclear and cytoplasmic ...
16889374 - Construction of a pichia pastoris cell-surface display system using flo1p anchor system.
424314 - Metabolic stability of 2' 5'oligo (a) and activity of 2' 5'oligo (a)-dependent endonucl...
17967934 - Evaluation of mammalian cell-free systems of nuclear disassembly and assembly.
Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  BioTechniques     Volume:  48     ISSN:  1940-9818     ISO Abbreviation:  BioTechniques     Publication Date:  2010 Feb 
Date Detail:
Created Date:  2010-04-02     Completed Date:  2010-06-28     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  8306785     Medline TA:  Biotechniques     Country:  England    
Other Details:
Languages:  eng     Pagination:  113-20     Citation Subset:  IM    
Affiliation:
Institute for Neurophysiology, University of Cologne, Cologne, Germany. akp59@uni-koeln.de
Export Citation:
APA/MLA Format     Download EndNote     Download BibTex
MeSH Terms
Descriptor/Qualifier:
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:
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


Previous Document:  Effect of Budesonide on TGF-beta1 enhanced VEGF Production by Lung Fibroblasts.
Next Document:  An agarose spot assay for chemotactic invasion.