FLUORESCENCE DETECTION OF DNA BREAKS USING MOLECULAR OSCILLATORS

摘要

A method to detect DNA breaks includes providing a mixture of fluorescence energy transfer molecular oscillators and a DNA sample. The FET oscillator is a synthetic oligonucleotide that has a topoisomerase recognition sequence, a fluorescence donor and a fluorescence acceptor. The synthetic oligonucleotide is bound to a type I topoisomerase capable of binding to the topoisomerase recognition sequence. The mixture is irradiated at a wavelength of the fluorescence donor, and the emission is measured. Another variant of the disclosure is a probe for detecting DNA breaks utilizing a synthetic oligonucleotide comprising a topoisomerase recognition sequence, a fluorescence donor, and a nonradiative fluorescence quencher. Yet another variant of the disclosure is a probe for detecting DNA breaks utilizing a synthetic oligonucleotide comprising a topoisomerase recognition sequence, a fluorescence donor, and a fluorescence acceptor. The mixture is irradiated at a wavelength of the fluorescence donor; and the emission is measured. A method to detect DNA breaks may use these probes in a manner similar to that of the FET oscillator. The FET oscillators and probes are capable of being prepared in a kit formulation.