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Related Experiment Video

Updated: Nov 1, 2025

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Chimeric human opsins as optogenetic light sensitisers.

Doron G Hickey1,2, Wayne I L Davies1,3,4, Steven Hughes1,5

  • 1Nuffield Laboratory of Ophthalmology, University of Oxford, Oxford, OX1 3QU, UK.

The Journal of Experimental Biology
|June 21, 2021
PubMed
Summary

Researchers engineered human opsin variants for optogenetics. These chimeric opsins show tunable G-protein coupling, expanding tools for cell signaling research and therapies.

Keywords:
ChimeraG proteinOpsinOptogeneticsPhototransductionRetina

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Area of Science:

  • Optogenetics
  • Molecular Biology
  • Cell Signaling

Background:

  • Human opsin-based photopigments are promising light-sensitizers but their functionality is limited by dependence on specific second messenger proteins in non-native cells.
  • Understanding opsin-G protein interactions is crucial for developing advanced optogenetic tools.

Purpose of the Study:

  • To engineer chimeric human opsins with modified intracellular domains to alter G-protein coupling specificity and functionality.
  • To investigate the impact of intracellular domain composition on opsin trafficking, chromophore dependence, and signaling pathway activation (Gi and Gq/11).

Main Methods:

  • Generation of eight chimeric human opsins using rhodopsin (RHO) or long-wavelength-sensitive (LWS) opsin backbones with intracellular domains from melanopsin.
  • Assessment of Gi and Gq/11 pathway activation in response to light.
  • Evaluation of chromophore dependence for opsin function.
  • Immunocytochemistry (ICC) to determine plasma membrane trafficking of chimeric opsins.

Main Results:

  • Rhodopsin/melanopsin chimeras coupled to both Gi and Gq/11 pathways, with increased melanopsin substitution enhancing Gq/11 activity and reducing Gi activation.
  • Rhodopsin and rhodopsin/melanopsin chimeras required exogenous chromophore, unlike wild-type melanopsin.
  • Wild-type LWS opsin and LWS/melanopsin chimeras showed minimal Gi activation and no detectable Gq/11 activation.
  • Increased melanopsin intracellular domains reduced plasma membrane trafficking.

Conclusions:

  • Gα coupling efficiency significantly influences the speed of cellular responses.
  • Engineered human opsins exhibit unique properties, offering expanded possibilities for customized optogenetic biotools in research and therapy.
  • The study highlights the importance of intracellular domain engineering for controlling opsin function and localization.