Protocol for detecting endogenous GPCR activity in primary cell cultures using ONE-GO biosensors
ONE vector G protein Optical (ONE-GO) biosensors can measure the activity of endogenously expressed G protein-coupled receptors (GPCRs) in primary cells. By detecting G proteins that belong to all four families (G<sub>s</sub>, G<sub>i/o</sub>, G<sub>q/11</sub>, G<sub>12/13</sub>) across cell types, these biosensors provide high experimental versatility. We first describe steps to express ONE-GO biosensors in primary cells using lentiviral transduction. We then detail how to carry out measurements and subsequent analysis to quantify changes in bioluminescence resonance energy transfer (BRET) reporting on endogenous GPCR activity. For complete details on the use and execution of this protocol, please refer to Janicot et al.<sup>1</sup>.
Authors
- Remi Janicot (ORCID: https://orcid.org/0000-0001-6907-7805)
- Mikel Garcia‐Marcos (ORCID: https://orcid.org/0000-0001-9513-4826)
Institutions
- Boston University (US)
Publication Details
- Journal
- STAR Protocols
- Published
- 2024-10-01
- DOI
- https://doi.org/10.1016/j.xpro.2024.103355
- Citations
- 1
- Primary Topic
- Receptor Mechanisms and Signaling
- Type
- article
- Field-Weighted Citation Impact
- 0.23
Funders
- American Heart Association
- National Institutes of Health
- National Institute of General Medical Sciences
- National Institute of Neurological Disorders and Stroke