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Oscillating Probe for Dual Detection of 5′PO<sub>4</sub> and 5′OH DNA Breaks in Tissue Sectionsстатья
- Автор: Didenko Vladimir V.
- Сборник: Fluorescent Energy Transfer Nucleic Acid Probes
- Год издания: 2006
- Место издания: Humana Press New Jersey
- Первая страница: 59
- Последняя страница: 70
- DOI: 10.1385/1-59745-069-3:59
- Аннотация: Several types of DNA cuts are used as markers of apoptosis for detection of apoptotic cells in situ. We recently introduced a ligase-based in situ assay that is specific for a single type of DNA damage--a double-strand break of DNase I-type, bearing 5'PO4. Here we describe a vaccinia topoisomerase I-based approach to label another type of DNA damage in situ--a double-strand break of DNase II-type, bearing 5'OH. The assay uses a new type of probe, a molecular oscillator. The probe self-assembles in solution out of a dual-hairpin oligonucleotide and vaccinia topoisomerase I. The enzyme continuously separates and religates two fluorescently labeled hairpins, which can participate in energy transfer. We describe the successful combination of topoisomerase-and ligase-based systems into an in situ assay. The assay uses an oscillating probe for simultaneous detection of two types of DNA cuts in tissue sections.
- Добавил в систему: Didenko Vladimir Vladimirovich