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  • Tetracycline-based system for controlled inducible expression of group III metabotropic glutamate receptors.

Tetracycline-based system for controlled inducible expression of group III metabotropic glutamate receptors.

Journal of biomolecular screening (2014-11-15)
Barbara Chruścicka, Grzegorz Burnat, Piotr Brański, Paulina Chorobik, Tomasz Lenda, Marcin Marciniak, Andrzej Pilc
ABSTRACT

A stable and inducible expression of metabotropic glutamate receptor type 4, 7, and 8 was obtained in T-REx 293 cells using the tetracycline system. Tetracycline administration to the cell medium resulted in rapid induction and time-dependent expression of mGlu receptors, which also correlates with its functionality in a cAMP accumulation assay. The pharmacological properties of recombinant mGlu receptors were verified using orthosteric and allosteric ligands. Data suggest that the Tet-on inducible system is suitable for functional mGlu receptors' expression and characterization by means of the cAMP accumulation assay. It makes this system a precise, reproducible, and large-scale screening method, as well as a reasonable tool to study signaling properties of mGlu receptors.

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GeneJuice® Transfection Reagent, Non-lipid based chemical transfection reagent optimized for maximum transfection efficiency, ease-of-use, and minimal cytotoxicity on a wide variety of mammalian cells.
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