Design of Environmentally Sensitive Fluorescent Nucleosides and Their Application to Innovative Fluorescence Probe for Gene Analysis

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We have designed a novel type of base-discriminating fluorescent (BDF) nucleosides which are used for SNP genotyping and gene detection. We also succeeded to design environmentally sensitive fluorescent (ESF) nucleosides that emit strong fluorescence at different wavelengths depending upon a difference of local environment near the ESF base. The newly designed ESF nucleosides may discriminate structural changes such as single strands, matches, mismatches and deletions when incorporated in DNA by a significant change of emission wavelengths as well as their intensities. Quencher and end free molecular beacons (MB) were discovered for the first time by using newly designed ESF nucleosides. These low cost MB were used for the gene detection in a cell. ESF nucleosides that are sensitive to pH were also designed. Such pH sensor ESF nucleoside that emits strong fluorescence only at bellow pH 5.5 was used for the detection of cancer cell.