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A Bioorthogonal Near-Infrared Fluorogenic Probe for mRNA Detection.
Haoxing Wu1, Seth C Alexander1, Shuaijiang Jin1
1Department of Chemistry and Biochemistry, University of California, San Diego , La Jolla, California 92093, United States.
Journal of the American Chemical Society
|August 12, 2016
Summary
Researchers developed a new bioorthogonal tetrazine uncaging reaction to visualize RNA in cells. This method uses vinyl ether caged fluorophores and nucleic acid probes for highly selective RNA detection, including in live cells with near-infrared dyes.
Area of Science:
- Biochemistry
- Molecular Biology
- Chemical Biology
Background:
- Developing methods for RNA visualization and detection is crucial.
- Bioorthogonal chemistry offers a promising avenue for nucleic acid imaging.
- Previous tetrazine reactions were limited in fluorogen diversity.
Purpose of the Study:
- To report a novel bioorthogonal tetrazine uncaging reaction.
- To expand the range of fluorogens activated by tetrazine reactions.
- To enable selective RNA detection in live cells.
Main Methods:
- Utilized tetrazine reactivity to unmask vinyl ether caged fluorophores.
- Conjugated fluorophores and tetrazine partners to antisense nucleic acid probes.
- Detected target RNA sequences in live cells using near-infrared emitting probes.
Main Results:
- Demonstrated a novel tetrazine uncaging reaction for fluorophore activation.
- Achieved highly selective fluorogenic reactivity of probes upon target RNA annealing.
- Successfully detected mRNA target sequences in live cells using NIR fluorogens.
Conclusions:
- The developed reaction expands tetrazine fluorogenic chemistry to NIR dyes.
- Proximity-induced tetrazine reactions offer broad utility for endogenous biomolecule imaging.
- This approach holds potential for visualizing nucleic acids in cells and tissues.
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