Probing PAC1 receptor activation across species with an engineered sensor.
basic_science · Level V
Where this comes from
- Record sourced from PubMed, PMID 39145773.
- Also identified by DOI 10.7554/eLife.96496 and PMC identifier 11326774.
- Licence recorded as CC BY.
- The licence permits redistribution, so the abstract is shown in full and the full text is available from the publisher.
Abstract
Class-B1 G-protein-coupled receptors (GPCRs) are an important family of clinically relevant drug targets that remain difficult to investigate via high-throughput screening and in animal models. Here, we engineered PAClight1<sub>P78A</sub>, a novel genetically encoded sensor based on a class-B1 GPCR (the human PAC1 receptor, hmPAC1R) endowed with high dynamic range (Δ<i>F</i>/<i>F</i><sub>0</sub> = 1100%), excellent ligand selectivity, and rapid activation kinetics (<i>τ</i><sub>ON</sub> = 1.15 s). To showcase the utility of this tool for in vitro applications, we thoroughly characterized and compared its expression, brightness and performance between PAClight1<sub>P78A</sub>-transfected and stably expressing cells. Demonstrating its use in animal models, we show robust expression and fluorescence responses upon exogenous ligand application ex vivo and in vivo in mice, as well as in living zebrafish larvae. Thus, the new GPCR-based sensor can be used for a wide range of applications across the life sciences empowering both basic research and drug development efforts.
Medical subject headings
- Zebrafish