Son Hayoung, Kim Dahham, Kim Sohee, Gi Byun Wan, Bum Park Seung
Department of Chemistry, Seoul National University, Seoul, 08826, Korea.
Angew Chem Int Ed Engl. 2025 Mar 17;64(12):e202421982. doi: 10.1002/anie.202421982. Epub 2024 Dec 13.
Tetrazine (Tz)-embedded fluorescent probes, known for their fluorogenicity following bioorthogonal inverse electron-demand Diels-Alder (iEDDA) reactions, are extensively used in bioimaging. Despite extensive research on fluorogenic Tz probes, there has been limited systematic exploration of their fluorogenic responses with various dienophiles. In this study, we elucidate the structure-fluorogenic property relationship of bioorthogonal Tz probes. We synthesized a series of Seoul-Fluor-Tz (SF) probes designed to exhibit differentiated turn-on fluorescence upon iEDDA reactions with three dienophiles: trans-cyclooctene (TCO), bicyclo[6.1.0]nonyne (BCN), and spiro[2.3]hex-1-ene (Sph). Our findings revealed that the fluorogenic properties of the SF probes are highly dependent on the structures of Tz-dienophile adducts. By systematically modifying the electronic properties and employing quantum chemical calculations, we developed a series of SF probes with optimal dienophile-dependent fluorescence. These probes enabled simultaneous dual-color imaging of different cellular targets using a single probe, providing a robust approach for advanced bioimaging applications that require precise and efficient multicolor labeling strategies.
四嗪（Tz）嵌入的荧光探针，因其在生物正交逆电子需求狄尔斯-阿尔德（iEDDA）反应后的荧光生成特性而闻名，被广泛应用于生物成像。尽管对荧光Tz探针进行了广泛研究，但对于它们与各种亲双烯体的荧光反应，尚未有系统的探索。在本研究中，我们阐明了生物正交Tz探针的结构-荧光特性关系。我们合成了一系列首尔荧光-Tz（SF）探针，这些探针设计用于在与三种亲双烯体：反式环辛烯（TCO）、双环[6.1.0]壬炔（BCN）和螺[2.3]己-1-烯（Sph）发生iEDDA反应时表现出不同的开启荧光。我们的研究结果表明，SF探针的荧光特性高度依赖于Tz-亲双烯体加合物的结构。通过系统地改变电子性质并采用量子化学计算，我们开发了一系列具有最佳亲双烯体依赖性荧光的SF探针。这些探针能够使用单个探针同时对不同的细胞靶点进行双色成像，为需要精确高效多色标记策略的先进生物成像应用提供了一种强大的方法。
