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- W4377029136 abstract "<b>Abstract ID 13744</b> <b>Poster Board 287</b> G protein-coupled receptors (GPCRs) are the largest family of druggable targets in the human genome. They initiate a plethora of signaling pathways, including responses to most neurotransmitters and two-thirds of hormones, primarily by activating heterotrimeric G-proteins (Gαβγ) inside the cell. Traditional approaches to measure GPCR activity rely on indirect reporters subject to amplification or cross-talk. Biosensors that directly measure G protein activity (i.e., formation of Gα-GTP or Gβγ dissociation) are not limited by these factors, but often require overexpression of exogenous G protein, which can affect endogenous signaling, and are not suitable for implementation in physiologically relevant models. We set out to overcome all these limitations by generating transgenic mouse lines for the conditional expression of biosensors that can detect the activity of endogenous G proteins in any primary cell type compatible with <i>in vitro</i> culture. These sensors are named BERKY, for BRET biosensor with ER/K linker and YFP. We designed the mouse lines based on previously described BERKY biosensors for free Gβγ, Gαq-GTP, Gα13-GTP, or Gαi-GTP, and now present progress on the development of a new BERKY biosensor for Gαs-GTP, which would ensure coverage of the detection of activity of all G protein families. We currently have mouse lines for four biosensor breeding in our facilities, and have validated for some of them that endogenous G proteins activity can be detected upon conditional, Cre-dependent expression of the biosensor in primary cells. Moreover, our results also indicate that the expression of this biosensor does not interfere with endogenous G protein signaling. In summary, we are developing a platform of transgenic mice that will allow to measure the endogenous activity of any G protein subtype in physiologically relevant cell types without affecting native G protein signaling." @default.
- W4377029136 created "2023-05-19" @default.
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- W4377029136 date "2023-05-18" @default.
- W4377029136 modified "2023-10-18" @default.
- W4377029136 title "Development of Transgenic Mouse Platform for the Detection of Endogenous G protein Activity" @default.
- W4377029136 doi "https://doi.org/10.1124/jpet.122.137440" @default.
- W4377029136 hasPublicationYear "2023" @default.
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