PT  - JOURNAL ARTICLE
AU  - Jan N. Hansen
AU  - Fabian Kaiser
AU  - Christina Klausen
AU  - Birthe Stüven
AU  - Raymond Chong
AU  - Wolfgang Bönigk
AU  - David U. Mick
AU  - Andreas Möglich
AU  - Nathalie Jurisch-Yaksi
AU  - Florian I. Schmidt
AU  - Dagmar Wachten
TI  - Nanobody-directed targeting of optogenetic tools to study signaling in the primary cilium
AID  - 10.1101/2020.02.04.933440
DP  - 2020 Jan 01
TA  - bioRxiv
PG  - 2020.02.04.933440
4099  - http://biorxiv.org/content/early/2020/04/15/2020.02.04.933440.short
4100  - http://biorxiv.org/content/early/2020/04/15/2020.02.04.933440.full
AB  - 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 and function has been challenging due to the lack of tools to temporarily manipulate and analyze ciliary signaling. Here, we describe a nanobodybased targeting approach for optogenetic tools that is applicable in vitro and in vivo and allows to specifically analyze ciliary signaling and function. Thereby, we overcome the loss of protein function observed after direct fusion to a ciliary targeting sequence. We functionally localized modifiers of cAMP signaling, i.e. the photo-activated adenylate cyclase bPAC and the light-activated phosphodiesterase LAPD, as well as 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.Competing Interest StatementThe authors have declared no competing interest.