GPCR-Based Genetically Encoded Fluorescent Indicators
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Updated: Oct 31, 2026

Genetic Barcoding with Fluorescent Proteins for Multiplexed Applications
Published on: April 14, 2015
Valentin Lu Rohner1, Ralph Gradisch1, Tommaso Patriarchi2,3
1Institute of Pharmacology and Toxicology, University of Zürich, Zürich, Switzerland.
Abstract:
GPCR-based genetically encoded fluorescent indicators enable the detection of a vast diversity of endogenous ligands at high spatiotemporal resolution and have been transforming neuroscientific research. The development and characterization of indicators based on different GPCR scaffolds have proven time-consuming and laborious. In this protocol we describe an alignment-based grafting approach to generate potential indicator candidates in a single cloning step and outline their evaluation using confocal fluorescence microscopy. To characterize the dose-response behavior of functional indicators, we describe the generation of stable HEK293T cell lines and their use in combination with a fluorescence microplate reader to determine the EC50 of a ligand.
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