A new tool to study signaling with the help of nanobodies

June 27, 2020

Schematic drawing for the nanobody approach by Jan Niklas Hansen

Zebrafish larva that expresses RFP everywhere without (top) and with (bottom) nanobody that pulls RFP into the cilium. The cilia are shown in cyan and the nanobody in green. Picture by Jan Niklas Hansen.

 

Combining optogenetics with nanobody technology

A new study of the groups from Dagmar Wachten and Florian I. Schmidt from the Institute of Innate Immunity shows the capability of combining two different techniques for studying unknown processes. The results were published in the Journal eLife. Using a nanobody-based targeting approach in combination with optogenetic tools could overcome the loss of protein function observed after fusion to ciliary targeting sequences. Hereby the ciliary signaling and function can be studied in mammalian cells an in vivo in zebrafish.

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. 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.

You can find the publication here.