Unveiling the Structure-Fluorogenic Property Relationship of Seoul-Fluor-Derived Bioorthogonal Tetrazine Probes

Area of Science:

• Bioorthogonal chemistry
• Fluorescent probe development
• Bioimaging applications

Background:

• Tetrazine (Tz)-based fluorescent probes are widely used in bioimaging due to their fluorogenicity via bioorthogonal inverse electron-demand Diels-Alder (iEDDA) reactions.
• Limited systematic studies exist on the structure-fluorogenic property relationship of these probes with various dienophiles.

Purpose of the Study:

• To elucidate the structure-fluorogenic property relationship of bioorthogonal tetrazine (Tz) probes.
• To develop novel Seoul-Fluor-Tz (SF_Tz) probes with optimized fluorescence responses.
• To enable simultaneous multicolor bioimaging applications.

Main Methods:

• Synthesis of a series of Seoul-Fluor-Tz (SF_Tz) probes.
• Evaluation of probe fluorogenicity upon reaction with three dienophiles: trans-cyclooctene (TCO), bicyclo[6.1.0]nonyne (BCN), and spiro[2.3]hex-1-ene (Sph).
• Systematic modification of electronic properties and application of quantum chemical calculations.

Main Results:

• The fluorogenic properties of SF_Tz probes were found to be highly dependent on the specific Tz-dienophile adduct structures.
• Optimized SF_Tz probes exhibiting distinct dienophile-dependent fluorescence were successfully developed.
• These probes facilitated simultaneous dual-color imaging of different cellular targets using a single probe platform.

Conclusions:

• A clear structure-fluorogenic property relationship was established for bioorthogonal Tz probes.
• The developed SF_Tz probes offer a versatile and robust platform for advanced multicolor bioimaging.
• This work provides a foundation for designing next-generation probes for precise cellular labeling strategies.