@inbook{1bf4a3ee89b84cf2b945e09383c73972,
title = "Automated Plate Reader-Based Assays of Light-Activated GPCRs",
abstract = "In the emerging field of optogenetics, light-sensitive G-protein coupled receptors (GPCRs) allow for the temporally precise control of canonical cell signaling pathways. Expressing, stimulating, and measuring the activity of light-sensitive GPCRs (e.g., opsins or chimeric OptoXRs) in mammalian cells is a nontrivial task as many standard assay practices are not compatible with light-sensitive molecular tools. In this chapter, we present a method for quantifying opsin activity in automated plate reader-based assays without the need for additional optical hardware (i.e., light sources). The protocol is applied to assess cAMP levels downstream of a chimeric OptoXR but can be expanded to other opsins and second messengers, such as Ca2+ mobilization. We describe how the internal optical components in commonly available plate readers can be utilized to both activate and detect kinetic and dose-response relationships, as well as provide general guidance for optimizing assays with light-sensitive molecular tools.",
keywords = "Automation, Cell signaling, Opsin, Optogenetics, OptoXR, Synthetic biology",
author = "Gerrard, {Elliot J.} and Alexandra-Madelaine Tichy and Harald Janovjak",
year = "2025",
doi = "10.1007/978-1-0716-4047-0_16",
language = "English",
isbn = "978-1-0716-4046-3",
series = "Methods in molecular biology (Clifton, N.J.)",
publisher = "Springer ",
pages = "217--229",
editor = "Armin Baumschlager",
booktitle = "Optogenetics",
}