Shrimp Rhodopsins as new far-red absorbing optogenetic tools
Participants:
1) Peter Hegemann (PI; Humboldt University Berlin), coworker:
2) Sonja Kleinlogel (PI; University of Bern, Switzerland), coworker:
Abstract:
Red light is most attractive for stimulation of optogenetic actuators and reporter systems, since it is less harmful than short-wavelength light, penetrates well through tissue and does spectrally not interfere with most other available optical tools. Among the animal rhodopsins with the highest sensitivity to red light in nature are the mantis shrimp’s red sensitive rhodopsins that can be activated with near-infrared light beyond 700 nm. We will characterize the 9 already sequenced rhodopsins of the Shrimp Neogonodactylus oerstedii and variants of related species. We will overcome the poor expression of invertebrate rhodopsin by codon optimization and tailoring of the C- and N-termini and characterize the biophysical properties of the recombinant proteins. We will determine and modulate the G-protein coupling and use these rhodopsins for the modification of signaling cascades in neurons for non-invasive deep tissue activation.
Logic of interactions:
Published ShrimpOPNs will be optimized for expression in HEK293 cells and later neurons by both, the Hegemann and Kleinlogel labs. The Hegemann lab will perform a detailed biophysiological characterization of selected ShrimpOPNs, whilst the Kleinlogel lab will determine and modify G-protein selectivity and validate non-invasive deep tissue activation in vivo by combined electrophysiological and behavioral experiments.
Publications Hegemann:
Ritter E, Puskar L, Bartl FJ, Aziz EF, Hegemann P, Schade U (2015) Time-resolved infrared spectroscopic techniques as applied to channelrhodopsin. Front Mol Biosci 2: 38.
Wietek J, Rodriguez-Rozada S, Tutas J, Tenedini F, Grimm C, Oertner TG, Soba P, Hegemann P, Wiegert JS. (2017) Anion-conducting channelrhodopsins with tuned spectra and modified kinetics engineered for optogenetic applications. Sci Rep. 2017 Nov 2;7(1):14957.
Vierock, J., Grimm, C., Nitzen, N., and Hegemann, P. (2017) Molecular determinants of proton selectivity and gating in the red-light activated channelrhodopsin Chrimson. Sci. Rep. 7(1):9928.
Oda, K., Vierock, J., Oishi, S., Rodriguez-Rozada, S., Taniguchi, R., Yamashita, K., Wiegert, J.S., Nishizawa, T., Hegemann, P., Nureki, O. (2018) Crystal structure of the red light-activated channelrhodopsin Chrimson. Nature Comm. 9(1):3949.
Publications Kleinlogel:
Kleinlogel S, Marshall NJ. Electrophysiological evidence for linear polarization sensitivity in the compound eyes of the stomatopod crustacean Gonodactylus chiragra. J Exp Biol. 2006 Nov;209(Pt 21):4262-72.
Marshall J, Cronin TW, Kleinlogel S. Stomatopod eye structure and function: a review. Arthropod Struct Dev. 2007 Dec;36(4):420-48.
Chiou TH, Kleinlogel, S., Cronin T, Caldwell R, Loeffler B, Siddiqi A, Goldizen, A.,Marshall J. Circular polarization vision in a stomatopod crustacean. Curr Biol. 2008 Mar 25;18(6):429-34.
Kleinlogel S, White AG.The secret world of shrimps: polarisation vision at its best. PLoS One. 2008 May 14;3(5):e2190.
Kleinlogel S (2016) Optogenetic user's guide to Opto-GPCRs. Front Biosci (Landmark Ed) 21: 794–805. SK6: van Wyk M, Pielecka-Fortuna J, Löwel S, Kleinlogel S (2015) Restoring the ON Switch in Blind Retinas: Opto-mGluR6, a Next-Generation, Cell-Tailored Optogenetic Tool. PLoS Biol 13:e1002143.