Abstract
Stimulation of G protein-coupled receptors induces an intracellular signaling cascade that activates transcription factors in the nucleus and thus ultimately leads to transcriptional changes. We have designed reporter genes that specifically measure the activities of the stimulus-responsive transcription factors AP-1, CREB, Egr-1, and serum response factor/ternary complex factor. The transcriptional activation potential of CREB and Elk-1 is measured using fusion proteins consisting of the phosphorylation-dependent activation domain of these transcription factors joined with the DNA-binding domain of the yeast transcription factor GAL4. Lentiviral gene transfer is used to implant the reporter genes into the chromatin of the cells, to ensure that the reporter genes are embedded into a nucleosomal structure. Recombinant lentiviruses infect mitotic and postmitotic cells such as neurons, thus allowing the analysis of transcriptional changes following the activation of G protein-coupled designer receptors.
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Rössler, O.G., Thiel, G. (2015). Regulation of Gene Transcription Following Stimulation of Gαq-Coupled Designer Receptors. In: Thiel, G. (eds) Designer Receptors Exclusively Activated by Designer Drugs. Neuromethods, vol 108. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-2944-3_3
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DOI: https://doi.org/10.1007/978-1-4939-2944-3_3
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