RT Journal Article
SR Electronic
T1 In vivo photocontrol of microtubule dynamics and integrity, migration and mitosis, by the potent GFP-imaging-compatible photoswitchable reagents SBTubA4P and SBTub2M
JF bioRxiv
FD Cold Spring Harbor Laboratory
SP 2021.03.26.437160
DO 10.1101/2021.03.26.437160
A1 Li Gao
A1 Joyce C.M. Meiring
A1 Adam Varady
A1 Iris E. Ruider
A1 Constanze Heise
A1 Maximilian Wranik
A1 Cecilia D. Velasco
A1 Jennifer A. Taylor
A1 Beatrice Terni
A1 Jörg Standfuss
A1 Clemens C. Cabernard
A1 Artur Llobet
A1 Michel O. Steinmetz
A1 Andreas R. Bausch
A1 Martin Distel
A1 Julia Thorn-Seshold
A1 Anna Akhmanova
A1 Oliver Thorn-Seshold
YR 2021
UL http://biorxiv.org/content/early/2021/03/27/2021.03.26.437160.abstract
AB Photoswitchable reagents to modulate microtubule stability and dynamics are an exciting tool approach towards micron- and millisecond-scale control over endogenous cytoskeleton-dependent processes. When these reagents are globally administered yet locally photoactivated in 2D cell culture, they can exert precise biological control that would have great potential for in vivo translation across a variety of research fields and for all eukaryotes. However, photopharmacology’s reliance on the azobenzene photoswitch scaffold has been accompanied by a failure to translate this temporally- and cellularly-resolved control to 3D models or to in vivo applications in multi-organ animals, which we attribute substantially to the metabolic liabilities of azobenzenes.Here, we optimised the potency and solubility of metabolically stable, druglike colchicinoid microtubule inhibitors based instead on the styrylbenzothiazole (SBT) photoswitch scaffold, that are non-responsive to the major fluorescent protein imaging channels and so enable multiplexed imaging studies. We applied these reagents to 3D systems (organoids, tissue explants) and classic model organisms (zebrafish, clawed frog) with one- and two-protein imaging experiments. We successfully used systemic treatment plus spatiotemporally-localised illuminations in vivo to photocontrol microtubule dynamics, network architecture, and microtubule-dependent processes in these systems with cellular precision and second-level resolution. These nanomolar, in vivo-capable photoswitchable reagents can prove a game-changer for high-precision cytoskeleton research in cargo transport, cell motility, cell division and development. More broadly, their straightforward design can also inspire the development of similarly capable optical reagents for a range of protein targets, so bringing general in vivo photopharmacology one step closer to productive realisation.Competing Interest StatementThe authors have declared no competing interest.