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Design, construction, and validation of optogenetic proteins
Colin P O'Banion1, Anwesha Goswami1, David S Lawrence2
1Division of Chemical Biology and Medicinal Chemistry, UNC Eshelman School of Pharmacy, University of North Carolina, Chapel Hill, NC, United States.
Methods in Enzymology
|May 27, 2019
Summary
This study introduces a new method to control cellular signaling using light. Researchers engineered proteins to become optogenetic tools, enabling precise manipulation of cell functions with light.
Area of Science:
- Cellular biology
- Molecular biology
- Biotechnology
Background:
- Cellular optogenetics uses light-controlled proteins to precisely regulate cellular signaling.
- Existing methods often require specific protein targets, limiting broader application.
Purpose of the Study:
- To develop a generalized method for converting any protein of interest (POI) into an optogenetic tool.
- To demonstrate the creation of three distinct optogenetic POIs using the Cry2-Cib system.
Main Methods:
- Engineered a constitutively active POI, independent of natural cellular regulation.
- Fused the modified POI to the Cry2-Cib photodimerizing system.
- Utilized light to recruit the optogenetic POI to targeted subcellular locations.
Main Results:
- Successfully generated three novel optogenetic proteins of interest.
- Demonstrated light-inducible recruitment of POIs to specific cellular compartments.
- Validated the ability to perturb cellular signaling with spatiotemporal precision.
Conclusions:
- The presented approach offers a versatile strategy for engineering optogenetic tools from diverse proteins.
- This method enhances the toolkit for cellular optogenetics, enabling new research possibilities.
- The developed optogenetic POIs can be used to study protein function and cellular pathways with light control.
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