Abstract
Circadian clocks are self-sustained and cell-autonomous oscillators. They respond to various extracellular cues depending on the time-of-day and the signal intensity. Phase Transition Curves (PTCs) are instrumental in uncovering the full repertoire of responses to a given signal. However, the current methodologies for reconstructing PTCs are low-throughput, laborious, and resource- and time-consuming. We report here the development of an efficient and high throughput assay, dubbed Circadian Single-Cell Oscillators PTC Extraction (Circa-SCOPE) for generating high-resolution PTCs. This methodology relies on continuous monitoring of single-cell oscillations to reconstruct a full PTC from a single culture, upon a one-time intervention. Using Circa-SCOPE, we characterize the effects of various pharmacological and blood-borne resetting cues, at high temporal resolution and a wide concentration range. Thus, Circa-SCOPE is a powerful tool for comprehensive analysis and screening for circadian clocks’ resetting cues, and can be valuable for basic as well as translational research.
| Original language | English |
|---|---|
| Article number | 5903 |
| Journal | Nature Communications |
| Volume | 12 |
| Issue number | 1 |
| DOIs | |
| State | Published - 1 Dec 2021 |
| Externally published | Yes |
Bibliographical note
Publisher Copyright:© 2021, The Author(s).
Funding
We are grateful to all the members of the Asher lab for their comments on the paper. We wish to thank Uriel Rosen (WIS IT unit) and Jonathan Sobel for their assistance in establishing the CellProfiler pipeline on the institutional cluster. G.A. is supported by a grant from the European Research Council (ERC-2017 CIRCOMMUNICATION 770869), Abisch Frenkel Foundation for the Promotion of Life Sciences, Adelis Foundation, Susan and Michael Stern.
| Funders | Funder number |
|---|---|
| Achelis Foundation | |
| H2020 European Research Council | |
| European Commission | ERC-2017 CIRCOMMUNICATION 770869 |
| Abisch-Frenkel-Stiftung |