We found U2R1,a previously discovered G-rich DNA sequence,could form the G-quadruplex(G4)structure at high tem-perature and maintain high stability.By utilizing its specific binding ability with the N-methylmesoporphy...We found U2R1,a previously discovered G-rich DNA sequence,could form the G-quadruplex(G4)structure at high tem-perature and maintain high stability.By utilizing its specific binding ability with the N-methylmesoporphyrin IX(NMM,a fluorescent probe),a thermophilic signal transduction unit was proposed,that was the NMM-U2R1 complex.Current methods for single nucleotide polymorphism(SNP)detection relying on the room-temperature rolling circle amplification system often suffered from poor accuracy,since the low temperature lowers the sensitivity for identifying the base mismatches.In this work,we combined the thermal stable signal transduction unit with the isothermal amplification reaction to develop a thermophilic fluorescent assay.High temperature could ensure the accuracy of base pairing.Based on this,this fluorescent assay has been successfully applied for the identification of one-or two-mismatched base DNA or microRNA 21.And it is expected to be generally applicable to identify SNPs in many other sequences.Furthermore,this work will open new opportunities for development of the thermally stable G4 DNA in biosensor.展开更多
基金financially supported by the National Natural Science Foundation of China (21922601, 51808096)Liaoning Revitalization Talents Program (XLYC1807080)the Fundamental Research Funds for the Central Universities (DUT20YG123)
文摘We found U2R1,a previously discovered G-rich DNA sequence,could form the G-quadruplex(G4)structure at high tem-perature and maintain high stability.By utilizing its specific binding ability with the N-methylmesoporphyrin IX(NMM,a fluorescent probe),a thermophilic signal transduction unit was proposed,that was the NMM-U2R1 complex.Current methods for single nucleotide polymorphism(SNP)detection relying on the room-temperature rolling circle amplification system often suffered from poor accuracy,since the low temperature lowers the sensitivity for identifying the base mismatches.In this work,we combined the thermal stable signal transduction unit with the isothermal amplification reaction to develop a thermophilic fluorescent assay.High temperature could ensure the accuracy of base pairing.Based on this,this fluorescent assay has been successfully applied for the identification of one-or two-mismatched base DNA or microRNA 21.And it is expected to be generally applicable to identify SNPs in many other sequences.Furthermore,this work will open new opportunities for development of the thermally stable G4 DNA in biosensor.
