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Circa-SCOPE: high-throughput live single-cell imaging method for analysis of circadian clock resetting

Gal Manella, Dan Aizik, Rona Aviram, Marina Golik and Gad Asher ()
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Gal Manella: Weizmann Institute of Science
Dan Aizik: Weizmann Institute of Science
Rona Aviram: Weizmann Institute of Science
Marina Golik: Weizmann Institute of Science
Gad Asher: Weizmann Institute of Science

Nature Communications, 2021, vol. 12, issue 1, 1-14

Abstract: 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.

Date: 2021
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DOI: 10.1038/s41467-021-26210-1

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