2.50
Hdl Handle:
http://hdl.handle.net/10033/153959
Title:
Rapid establishment of G-protein-coupled receptor-expressing cell lines by site-specific integration.
Authors:
Schucht, Roland; Lydford, Simon; Andzinski, Lisa; Zauers, Jeannette; Cooper, James; Hauser, Hansjörg; Wirth, Dagmar; May, Tobias
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.
Affiliation:
Department of Gene Regulation and Differentiation, HZI-Helmholtz Centre for Infection Research, Braunschweig, Germany. roland.schucht@inscreenex.com
Citation:
Rapid establishment of G-protein-coupled receptor-expressing cell lines by site-specific integration. 2011, 16 (3):323-31 J Biomol Screen
Journal:
Journal of biomolecular screening
Issue Date:
Mar-2011
URI:
http://hdl.handle.net/10033/153959
DOI:
10.1177/1087057110396371
PubMed ID:
21335600
Type:
Article
Language:
en
ISSN:
1552-454X
Appears in Collections:
Publications of Dept. Gene Regulation and Differentiation (RDIF)

Full metadata record

DC FieldValue Language
dc.contributor.authorSchucht, Rolanden
dc.contributor.authorLydford, Simonen
dc.contributor.authorAndzinski, Lisaen
dc.contributor.authorZauers, Jeannetteen
dc.contributor.authorCooper, Jamesen
dc.contributor.authorHauser, Hansjörgen
dc.contributor.authorWirth, Dagmaren
dc.contributor.authorMay, Tobiasen
dc.date.accessioned2011-10-26T12:38:25Z-
dc.date.available2011-10-26T12:38:25Z-
dc.date.issued2011-03-
dc.identifier.citationRapid establishment of G-protein-coupled receptor-expressing cell lines by site-specific integration. 2011, 16 (3):323-31 J Biomol Screenen
dc.identifier.issn1552-454X-
dc.identifier.pmid21335600-
dc.identifier.doi10.1177/1087057110396371-
dc.identifier.urihttp://hdl.handle.net/10033/153959-
dc.description.abstractThe 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.en
dc.language.isoenen
dc.subject.meshAnimalsen
dc.subject.meshCHO Cellsen
dc.subject.meshCell Lineen
dc.subject.meshCricetinaeen
dc.subject.meshCricetulusen
dc.subject.meshGene Expression Regulationen
dc.subject.meshGene Orderen
dc.subject.meshGene Targetingen
dc.subject.meshGenetic Vectorsen
dc.subject.meshHigh-Throughput Screening Assaysen
dc.subject.meshReceptors, G-Protein-Coupleden
dc.subject.meshRecombinasesen
dc.titleRapid establishment of G-protein-coupled receptor-expressing cell lines by site-specific integration.en
dc.typeArticleen
dc.contributor.departmentDepartment of Gene Regulation and Differentiation, HZI-Helmholtz Centre for Infection Research, Braunschweig, Germany. roland.schucht@inscreenex.comen
dc.identifier.journalJournal of biomolecular screeningen

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