title: Rapid establishment of G-protein-coupled receptor-expressing cell lines by site-specific integration.
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.
authors.affiliation: Department of Gene Regulation and Differentiation, HZI-Helmholtz Centre for Infection Research, Braunschweig, Germany. roland.schucht@inscreenex.com
authors.forename: Roland
authors.initials: R
authors.lastname: Schucht
authors.forename: Simon
authors.initials: S
authors.lastname: Lydford
authors.forename: Lisa
authors.initials: L
authors.lastname: Andzinski
authors.forename: Jeannette
authors.initials: J
authors.lastname: Zauers
authors.forename: James
authors.initials: J
authors.lastname: Cooper
authors.forename: Hansjörg
authors.initials: H
authors.lastname: Hauser
authors.forename: Dagmar
authors.initials: D
authors.lastname: Wirth
authors.forename: Tobias
authors.initials: T
authors.lastname: May
mesh_terms: D000818:Animals; D016466:CHO Cells; D002460:Cell Line; D006224:Cricetinae; D003412:Cricetulus; D005786:Gene Expression Regulation; D023061:Gene Order; D018390:Gene Targeting; D005822:Genetic Vectors; D057166:High-Throughput Screening Assays; D043562:Receptors, G-Protein-Coupled / Q000235:genetics* / Q000378:metabolism*; D045522:Recombinases / Q000378:metabolism
chemical_list: D043562:Receptors, G-Protein-Coupled; D045522:Recombinases