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Updated: Sep 9, 2025
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Long-range Channelrhodopsin-assisted Circuit Mapping of Inferior Colliculus Neurons with Blue and Red-shifted Channelrhodopsins
Published on: February 7, 2020
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Screening channelrhodopsins using robotic intracellular electrophysiology and single cell sequencing
Samuel Ehrlich1, Alexandra D VandeLoo2, Mohamed Badawy1,3
1George W. Woodruff School of Mechanical Engineering, Georgia Institute of Technology, Atlanta, Georgia, United States.
Biorxiv : the Preprint Server for Biology
|September 2, 2025
Summary
We developed a novel high-throughput method to link opsin genetic sequences with their functions, enabling faster engineering of these important proteins. This technique identified a mutation that disables ChrimsonR
Area of Science:
- Optogenetics
- Molecular Biology
- Neuroscience
Background:
- Engineering opsins is hindered by limited understanding of sequence-function relationships.
- The extensive opsin sequence space complicates systematic exploration.
Purpose of the Study:
- To develop a novel method for screening channel-rhodopsins.
- To create datasets linking opsin genetic sequence to function.
- To facilitate opsin engineering and understanding of sequence-function relationships.
Main Methods:
- Integrated robotic intracellular electrophysiology (Patch) for measuring optogenetic properties (Excite).
- Automated cell harvesting (Pick) for cells of interest.
- Subsequent genetic sequencing (Sequence) to link function to sequence.
Main Results:
- Sequenced over 50 cells with associated functional characterization.
- Demonstrated utility on heterogeneous opsin populations and single point mutations.
- Identified C160W mutation ablating ChrimsonR's light response.
Conclusions:
- The developed method offers efficient, standardized, and scalable characterization of opsin libraries.
- Enables large-scale datasets for advancing opsin engineering and sequence-function understanding.
- Outperforms traditional manual patch clamp screening in throughput and efficiency.
