Using the Flp-In™ T-Rex™ system to regulate GPCR expression

Methods Mol Biol. 2011:746:21-37. doi: 10.1007/978-1-61779-126-0_2.

Abstract

The development of a cell-based system that allows the integration of a gene of interest (GOI), such as a G protein-coupled receptor (GPCR), into a specific site on the genome, has made the generation of mammalian cell lines able to express such proteins easy and efficient. Flp-In™ stable cell lines are isogenic and hence protein expression is constant across a population of cells. A useful addition to the Flp-In™ system (Flp-In™ T-Rex™) allows this expression to be controlled by the addition of a small molecule inducer to the cell culture medium. Stable cell lines generated as described here can be used to great advantage in the study of receptor pharmacology signalling and oligomerisation.

MeSH terms

  • Clone Cells / drug effects
  • Clone Cells / metabolism
  • DNA Nucleotidyltransferases / metabolism*
  • Gene Expression Regulation* / drug effects
  • Genetic Techniques*
  • Genetic Vectors / genetics
  • Genetic Vectors / metabolism
  • HEK293 Cells
  • Humans
  • Receptors, G-Protein-Coupled / genetics*
  • Receptors, G-Protein-Coupled / metabolism*
  • Repressor Proteins / metabolism*
  • Tetracycline / pharmacology
  • Transcriptional Activation / drug effects
  • Transfection

Substances

  • Receptors, G-Protein-Coupled
  • Repressor Proteins
  • tetracycline resistance-encoding transposon repressor protein
  • DNA Nucleotidyltransferases
  • FLP recombinase
  • Tetracycline