Fluorescence-based detection of single nucleotide permutation in DNA via catalytically templated reaction

摘要

Templated reduction of low fluorescence azidocoumarin-PNA conjugate to high fluorescence aminocoumarin was achieved using a catalytic amount of DNA with single nucleotide resolution. Oligonucleotide-templated reactions have attracted tremendous interest for their implication in prebiotic chemistry and as a means to program and orchestrate designed reactions. An important application of such reactions is the sequence specific detection of DNA and RNA which is of paramount importance in biomedical research. For oligonucleotide detection, it is desirable to have a reaction where the template can turnover so to provide an amplification of the DNA or RNA signal with a single nucleotide resolution. Several strategies have already been reported notably based on autoligation, hydrolysis, the Staudinger reaction and chemical ligations; however, few methods were demonstrated to be catalytic in the template. Recently Seitz and Grossmann reported a system based on a transthioesterification to transfer a quencher group from one sequence to another with good turnover; however, the impact of mutations was only reported for a pyrimidine to purine mutation.