Live-cell imaging of circadian clock protein dynamics in CRISPR-generated knock-in cells

Christian H. Gabriel, Marta Olmo, Amin Zehtabian, Marten Jäger, Silke Reischl, Hannah Dijk, Carolin Ulbricht, Asylkhan Rakhymzhan, Thomas Korte, Barbara Koller, Astrid Grudziecki, Bert Maier, Andreas Herrmann, Raluca Niesner, Tomasz Zemojtel, Helge Ewers, Adrián E. Granada, Hanspeter Herzel and Achim Kramer ()
Additional contact information
Christian H. Gabriel: Charité Universitätsmedizin Berlin, corporate member of Freie Universität Berlin, Humboldt-Universität zu Berlin, and Berlin Institute of Health, Laboratory of Chronobiology
Marta Olmo: Institute for Theoretical Biology, Charité Universitätsmedizin Berlin
Amin Zehtabian: Institute for Chemistry and Biochemistry, Department of Biology, Chemistry and Pharmacy, Freie Universität Berlin
Marten Jäger: Berlin Institute of Health (BIH) Core Genomics Facility, Charité Universitätsmedizin Berlin
Silke Reischl: Charité Universitätsmedizin Berlin, corporate member of Freie Universität Berlin, Humboldt-Universität zu Berlin, and Berlin Institute of Health, Laboratory of Chronobiology
Hannah Dijk: Charité Universitätsmedizin Berlin, corporate member of Freie Universität Berlin, Humboldt-Universität zu Berlin, and Berlin Institute of Health, Laboratory of Chronobiology
Carolin Ulbricht: Immune Dynamics, Rheumatology and Clinical Immunology, Charité – Universitätsmedizin Berlin
Asylkhan Rakhymzhan: Biophysical Analytics, Deutsches Rheuma-Forschungszentrum (DRFZ), a Leibniz Institute
Thomas Korte: Molecular Biophysics, Department of Biology, Humboldt Universität zu Berlin
Barbara Koller: Charité Universitätsmedizin Berlin, corporate member of Freie Universität Berlin, Humboldt-Universität zu Berlin, and Berlin Institute of Health, Laboratory of Chronobiology
Astrid Grudziecki: Charité Universitätsmedizin Berlin, corporate member of Freie Universität Berlin, Humboldt-Universität zu Berlin, and Berlin Institute of Health, Laboratory of Chronobiology
Bert Maier: Charité Universitätsmedizin Berlin, corporate member of Freie Universität Berlin, Humboldt-Universität zu Berlin, and Berlin Institute of Health, Laboratory of Chronobiology
Andreas Herrmann: Molecular Biophysics, Department of Biology, Humboldt Universität zu Berlin
Raluca Niesner: Biophysical Analytics, Deutsches Rheuma-Forschungszentrum (DRFZ), a Leibniz Institute
Tomasz Zemojtel: Berlin Institute of Health (BIH) Core Genomics Facility, Charité Universitätsmedizin Berlin
Helge Ewers: Institute for Chemistry and Biochemistry, Department of Biology, Chemistry and Pharmacy, Freie Universität Berlin
Adrián E. Granada: Charité Comprehensive Cancer Center, Charité Universitätsmedizin Berlin
Hanspeter Herzel: Institute for Theoretical Biology, Charité Universitätsmedizin Berlin
Achim Kramer: Charité Universitätsmedizin Berlin, corporate member of Freie Universität Berlin, Humboldt-Universität zu Berlin, and Berlin Institute of Health, Laboratory of Chronobiology

Nature Communications, 2021, vol. 12, issue 1, 1-15

Abstract: Abstract The cell biology of circadian clocks is still in its infancy. Here, we describe an efficient strategy for generating knock-in reporter cell lines using CRISPR technology that is particularly useful for genes expressed transiently or at low levels, such as those coding for circadian clock proteins. We generated single and double knock-in cells with endogenously expressed PER2 and CRY1 fused to fluorescent proteins allowing us to simultaneously monitor the dynamics of CRY1 and PER2 proteins in live single cells. Both proteins are highly rhythmic in the nucleus of human cells with PER2 showing a much higher amplitude than CRY1. Surprisingly, CRY1 protein is nuclear at all circadian times indicating the absence of circadian gating of nuclear import. Furthermore, in the nucleus of individual cells CRY1 abundance rhythms are phase-delayed (~5 hours), and CRY1 levels are much higher (>5 times) compared to PER2 questioning the current model of the circadian oscillator.

Date: 2021
References: Add references at CitEc
Citations: View citations in EconPapers (1)

Downloads: (external link)
https://www.nature.com/articles/s41467-021-24086-9 Abstract (text/html)

Related works:
This item may be available elsewhere in EconPapers: Search for items with the same title.

Export reference: BibTeX RIS (EndNote, ProCite, RefMan) HTML/Text

Persistent link: https://EconPapers.repec.org/RePEc:nat:natcom:v:12:y:2021:i:1:d:10.1038_s41467-021-24086-9

Ordering information: This journal article can be ordered from
https://www.nature.com/ncomms/

DOI: 10.1038/s41467-021-24086-9

Access Statistics for this article

Nature Communications is currently edited by Nathalie Le Bot, Enda Bergin and Fiona Gillespie

More articles in Nature Communications from Nature
Bibliographic data for series maintained by Sonal Shukla () and Springer Nature Abstracting and Indexing ().