Nanobody-directed targeting of optogenetic tools to study signaling in the primary cilium

Title data

Hansen, Jan Niklas ; Kaiser, Fabian ; Klausen, Christina ; Stüven, Birthe ; Chong, Raymond ; Bönigk, Wolfgang ; Mick, David U. ; Möglich, Andreas ; Jurisch-Yaksi, Nathalie ; Schmidt, Florian I. ; Wachten, Dagmar:
Nanobody-directed targeting of optogenetic tools to study signaling in the primary cilium.
In: eLife. Vol. 9 (2020) . - e57907.
ISSN 2050-084X
DOI: https://doi.org/10.7554/eLife.57907

Official URL:

Abstract in another language

Compartmentalization of cellular signaling forms the molecular basis of cellular behavior. The primary cilium constitutes a subcellular compartment that orchestrates signal transduction independent from the cell body. Ciliary dysfunction causes severe diseases, termed ciliopathies. Analyzing ciliary signaling has been challenging due to the lack of tools investigate ciliary signaling. Here, we describe a nanobody-based targeting approach for optogenetic tools in mammalian cells and in vivo in zebrafish to specifically analyze ciliary signaling and function. Thereby, we overcome the loss of protein function observed after fusion to ciliary targeting sequences. We functionally localized modifiers of cAMP signaling, the photo-activated adenylate cyclase bPAC and the light-activated phosphodiesterase LAPD, and the cAMP biosensor mlCNBD-FRET to the cilium. Using this approach, we studied the contribution of spatial cAMP signaling in controlling cilia length. Combining optogenetics with nanobody-based targeting will pave the way to the molecular understanding of ciliary function in health and disease.

Further data

Item Type:	Article in a journal
Refereed:	Yes
Institutions of the University:	Faculties > Faculty of Biology, Chemistry and Earth Sciences > Department of Chemistry > Chair Biochemistry > Chair Biochemistry - Univ.-Prof. Dr. Andreas Möglich Faculties Faculties > Faculty of Biology, Chemistry and Earth Sciences Faculties > Faculty of Biology, Chemistry and Earth Sciences > Department of Chemistry Faculties > Faculty of Biology, Chemistry and Earth Sciences > Department of Chemistry > Chair Biochemistry
Result of work at the UBT:	Yes
DDC Subjects:	500 Science > 570 Life sciences, biology
Date Deposited:	25 Jun 2020 06:36
Last Modified:	12 Apr 2024 07:49
URI:	https://eref.uni-bayreuth.de/id/eprint/55613