Publication Database

Live-cell imaging of circadian clock protein dynamics in CRISPR-generated knock-in cells (2021)

Art

Zeitschriftenartikel / wissenschaftlicher Beitrag

Autoren

Gabriel, Christian H.

Del Olmo, Marta

Zehtabian, Amin

Jäger, Marten

Reischl, Silke

van Dijk, Hannah

Ulbricht, Carolin

Rakhymzhan, Asylkhan

Korte, Thomas

Koller, Barbara

Grudziecki, Astrid

Maier, Bert

Herrmann, Andreas

Niesner, Raluca (WE 2)

Zemojtel, Tomasz

Ewers, Helge

Granada, Adrián E.

Herzel, Hanspeter

Kramer, Achim

Quelle

Nature Communications

Bandzählung: 12

Heftzählung: 1

Seiten: Article number: 3796

ISSN: 2041-1723

Sprache

Englisch

Verweise

URL (Volltext): https://www.nature.com/articles/s41467-021-24086-9

DOI: 10.1038/s41467-021-24086-9

Pubmed: 34145278

Kontakt

Institut für Veterinär-Physiologie

Oertzenweg 19 b
14163 Berlin
+49 30 838 62600
physiologie@vetmed.fu-berlin.de

Abstract / Zusammenfassung

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.