Jing Nannan, Qin Yantian, Fan Xinli, Wang Qian, Wang Jing, You Fuping, Tang Xinjing
School of Basic Medical Sciences, Peking University, No. 38 Xueyuan Rd., Beijing 100191, China.
State Key Laboratory of Natural and Biomimetic Drugs, School of Pharmaceutical Sciences, Peking University, No. 38 Xueyuan Rd., Beijing 100191, China.
Biomolecules. 2025 May 23;15(6):752. doi: 10.3390/biom15060752.
DNA phosphorothioate (PT) modifications, characterized by the replacement of a non-bridging phosphate oxygen atom with a sulfur atom, are widely observed in bacterial genomes. Sensitive detection of phosphorothioate is crucial for elucidating their biological roles and functions. Herein, we developed an innovative method that leverages oligonucleotide-templated reactions (OTRs) and fluorogenic oligonucleotide probes. By optimizing temperature, probe sequence length, and the relative distance between PT position and the fluorophore probe, we achieved sensitive detection for DNA PT modifications through fluorogenic signal amplification, which provides an efficient and cost-effective approach for sensitive detection of phosphorothioate-modified DNA.
DNA硫代磷酸酯（PT）修饰的特征是用硫原子取代非桥连磷酸氧原子，在细菌基因组中广泛存在。硫代磷酸酯的灵敏检测对于阐明其生物学作用和功能至关重要。在此，我们开发了一种创新方法，该方法利用寡核苷酸模板反应（OTR）和荧光寡核苷酸探针。通过优化温度、探针序列长度以及PT位置与荧光团探针之间的相对距离，我们通过荧光信号放大实现了对DNA PT修饰的灵敏检测，这为硫代磷酸酯修饰的DNA的灵敏检测提供了一种高效且经济高效的方法。
