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Rapid establishment of G-protein-coupled receptor-expressing cell lines by site-specific integration.
Schucht, Roland Lydford, Simon Andzinski, Lisa Zauers, Jeannette Cooper, James Hauser, Hansjörg Wirth, Dagmar May, Tobias
Schucht, Roland
Lydford, Simon
Andzinski, Lisa
Zauers, Jeannette
Cooper, James
Hauser, Hansjörg
Wirth, Dagmar
May, Tobias
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2011-03
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Abstract
The establishment of mammalian cell lines reliably expressing G-protein-coupled receptors (GPCRs) can be a tedious and often time-consuming process. A strategy has been developed to allow the rapid production of such cell lines. The first step of this approach was the generation of a specialized master cell line, characterized by optimized stable expression of a membrane-bound reporter protein. In the second step, this reporter gene was exchanged for that of the GPCR of interest by a DNA recombinase "cut-and-paste" engineering step. It has been demonstrated that the resulting GPCR cell lines inherit the advantages of the master cell line, expressing the GPCR in a homogeneous and stable manner. The case studies presented demonstrate the functionality of the established GPCR cell lines, and most important, because of the highly efficient integration event, these recombinant GPCR-expressing cell lines were generated within a timeframe of 2 to 4 weeks. The advantages of this cut-and-paste approach versus other strategies such as Flp-In or Jump-In are compared.
Citation
Rapid establishment of G-protein-coupled receptor-expressing cell lines by site-specific integration. 2011, 16 (3):323-31 J Biomol Screen
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Article
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en
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1552-454X