2011•Methods in molecular biologyRequires access

Inducible Expression of G Protein-Coupled Receptors in Transfected Cells

Beryl Koener, Emmanuel Hermans

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Abstract

Biochemical or pharmacological studies of G protein-coupled receptors (GPCRs) are widely conducted in transfected mammalian cells. A variety of commercially available systems allow the generation of stable cell-lines in which expression of the recombinant receptor can be induced on addition of a defined chemical to the culture medium, which operates as a control switch for the transcription of the cloned sequence. Such systems offer the possibility to induce graded levels of receptor expression in the experimental model, or to induce an abrupt downregulation of receptor expression during the maintenance of the cell-line. This chapter provides an overview of the different systems available and provides methods for the generation and validation of stably transfected cell-lines expressing the GPCR of choice.

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What this paper is about

Biochemical or pharmacological studies of G protein-coupled receptors (GPCRs) are widely conducted in transfected mammalian cells. A variety of commercially available systems allow the generation of stable cell-lines in which expression of the recombinant receptor can be induced on addition of a defined chemical to the culture medium, which operates as a control switch for the transcription of the cloned sequence. Such systems offer the possibility to induce graded levels of receptor expression in the experimental model, or to induce an abrupt downregulation of receptor expression during the maintenance of the cell-line. This chapter provides an overview of the different systems available and provides methods for the generation and validation of stably transfected cell-lines expressing the GPCR of choice.

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Available abstract

Biochemical or pharmacological studies of G protein-coupled receptors (GPCRs) are widely conducted in transfected mammalian cells. A variety of commercially available systems allow the generation of stable cell-lines in which expression of the recombinant receptor can be induced on addition of a defined chemical to the culture medium, which operates as a control switch for the transcription of the cloned sequence. Such systems offer the possibility to induce graded levels of receptor expression in the experimental model, or to induce an abrupt downregulation of receptor expression during the maintenance of the cell-line. This chapter provides an overview of the different systems available and provides methods for the generation and validation of stably transfected cell-lines expressing the GPCR of choice.

Key concepts: Transfection, Receptor, G protein-coupled receptor, Cell biology, Cell culture, Downregulation and upregulation, Biology, Cell

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