A Bifunctional Fluorogenic Rhodamine Probe for Proximity-Induced Bioorthogonal Chemistry

Area of Science:

• Bioorthogonal chemistry
• Chemical biology
• Molecular imaging

Background:

• Bioorthogonal reactions are crucial tools in life sciences.
• Inverse electron demand Diels-Alder (DA_inv) reactions offer fast kinetics.
• Highly reactive dienophiles are unstable and prone to side reactions.

Purpose of the Study:

• To evaluate the acceleration of unreactive but stable dienophiles using DNA-templated proximity.
• To develop a novel bifunctional fluorogenic tetrazine rhodamine probe for kinetic studies.
• To identify a minimally sized fluorogenic tetrazine dienophile reactant pair for bioimaging.

Main Methods:

• Developed a modular synthetic route for a novel bifunctional fluorogenic tetrazine rhodamine probe.
• Utilized a fluorescence assay to determine reaction kinetics of various dienophiles.
• Investigated reaction rates under both proximity-driven and non-induced conditions.

Main Results:

• DNA-templated proximity accelerated the reaction by several orders of magnitude.
• Observed minimal background reaction when proximity was not induced.
• Identified a stable and minimally sized fluorogenic tetrazine dienophile reactant pair.

Conclusions:

• DNA-templated proximity is a powerful strategy to enhance the kinetics of stable dienophiles in bioorthogonal reactions.
• The developed probe and reactant pair show potential for precise visualization of biomolecular interactions.
• This approach offers temporal and spatial resolution for applications in living systems.