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Engineering and Application of LOV2-Based Photoswitches
S P Zimmerman1, B Kuhlman1, H Yumerefendi1
1University of North Carolina Chapel Hill, Chapel Hill, NC, United States.
Methods in Enzymology
|September 3, 2016
Summary
Cellular optogenetic switches, engineered from plant and bacterial photoreceptors, offer new ways to study biology. This work focuses on the LOV2 domain for designing effective light-controlled biological tools.
Area of Science:
- Biochemistry
- Molecular Biology
- Biophysics
Background:
- Optogenetic switches are novel biological tools enabling the study of previously intractable biological phenomena.
- Existing optogenetic tools are engineered from plant and bacterial photoreceptors, repurposing light-induced conformational changes.
Purpose of the Study:
- To examine the biophysical characteristics of the light-oxygen-voltage 2 (LOV2) domain from Avena sativa phototropin 1 for optogenetic switch design.
- To summarize previous design efforts and provide guidelines for effective optogenetic switch engineering.
- To present protocols for validating and improving newly designed photoswitches.
Main Methods:
- Focus on the biophysical characterization of the LOV2 domain.
- Review of existing optogenetic switch design strategies.
- Protocols for fluorescence polarization, phage display, and microscopy assays.
Main Results:
- The LOV2 domain's biophysical characteristics provide a strong foundation for engineering photoswitches.
- Established guidelines for effective optogenetic switch design based on the LOV2 domain.
- Optimized protocols for validating and improving photoswitch performance.
Conclusions:
- The LOV2 domain is a promising building block for creating novel cellular optogenetic switches.
- This work facilitates the development and application of light-controlled biological tools.
- Standardized assays are crucial for the advancement of optogenetics research.
Keywords:
Blue lightLOV2OptogeneticsPeptide cagingPhage displayPhotoswitchesProtein designProtein engineeringMore Related Videos
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