A Convenient Fluorogenic Detection Strategy for Phosphorothioate Modification of DNA Through Photocatalytic Oligonucleotide-Templated Reaction

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.