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Isomeric Tuning Yields Bright and Targetable Red Ca2+ Indicators
Claire Deo1, Shu-Hsien Sheu1, Jinyoung Seo1
1Janelia Research Campus , Howard Hughes Medical Institute , Ashburn , Virginia 20147 , United States.
Journal of the American Chemical Society
|August 21, 2019
Summary
Researchers created a new calcium (Ca2+) sensor for biological imaging. This bright and sensitive indicator uses a HaloTag system for cell-specific targeting, enabling visualization of calcium fluxes in the primary cilium.
Area of Science:
- Biochemistry
- Molecular Biology
- Cell Biology
Background:
- Genetically encoded protein tags enable targeted delivery of small-molecule fluorescent indicators for biological imaging.
- Optimizing these hybrid sensors requires redesigning synthetic indicators for cell-specific targeting without compromising probe performance.
Purpose of the Study:
- To develop a bright and sensitive calcium (Ca2+) indicator.
- To enable cell-specific targeting of the indicator using the HaloTag self-labeling tag system.
- To visualize Ca2+ fluxes in the primary cilium.
Main Methods:
- Systematic exploration of the relative configuration of dye and chelator in small-molecule Ca2+ indicators.
- Development of an
- isomeric tuning
- approach for indicator optimization.
- Utilizing the HaloTag self-labeling tag system for targeted delivery.
Main Results:
- Developed a bright and sensitive Ca2+ indicator with optimized dye-chelator configuration.
- Achieved cell-specific targeting of the indicator via the HaloTag system.
- Created a far-red targetable indicator capable of visualizing Ca2+ fluxes in the primary cilium.
Conclusions:
- The
- isomeric tuning
- approach is generalizable for designing targeted fluorescent indicators.
- The developed Ca2+ indicator facilitates advanced biological imaging, particularly in organelles like the primary cilium.
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