Chimeric human opsins as optogenetic light sensitisers

Human opsin-based photopigments have great potential as light-sensitisers, but their requirement for phototransduction cascade-specific second messenger proteins may restrict their functionality in non-native cell types. In this study, eight chimeric human opsins were generated consisting of a backbone of either a rhodopsin (RHO) or long-wavelength-sensitive (LWS) opsin and intracellular domains from G(q/11)-coupled human melanopsin. Rhodopsin/melanopsin chimeric opsins coupled to both Gi and G(q/11) pathways. Greater substitution of the intracellular surface with corresponding melanopsin domains generally showed greater G(q/11) activity with a decrease in Gi activation. Unlike melanopsin, rhodopsin and rhodopsin/melanopsin chimeras were dependent upon exogenous chromophore to function. By contrast, wild-type LWS opsin and LWS opsin/melanopsin chimeras showed only weak Gi activation in response to light, whilst G(q/11) pathway activation was not detected. Immunocytochemistry (ICC) demonstrated that chimeric opsins with more intracellular domains of melanopsin were less likely to be trafficked to the plasma membrane. This study demonstrates the importance of Ga coupling efficiency to the speed of cellular responses and created human opsins with a unique combination of properties to expand the range of customised optogenetic biotools for basic research and translational therapies.

Davies, Wayne I. L.Umeå universitet,Umeå centrum för molekylär medicin (UCMM),Nuffield Laboratory of Ophthalmology, University of Oxford, UK; 4 School of Life Sciences, College of Science, Health and Engineering, La Trobe University, Melbourne, Australia(Swepub:umu)wada0005 (författare)
Hughes, Steven (författare)
Rodgers, Jessica (författare)
Thavanesan, Navamayooran (författare)
MacLaren, Robert E. (författare)
Hankins, Mark W. (författare)
Umeå universitetUmeå centrum för molekylär medicin (UCMM) (creator_code:org_t)

Ingår i:Journal of Experimental BiologyCambridge : Company of Biologists LTD224:140022-09491477-9145

Av författaren/redakt...: Hickey, Doron G.; Davies, Wayne I. ...; Hughes, Steven; Rodgers, Jessica; Thavanesan, Nava ...; MacLaren, Robert ...; visa fler...; Hankins, Mark W.; visa färre...

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