title: Detection of DNA damage by thiazole orange fluorescence probe assisted with exonuclease III.
abstract: This work reports a fluorescent dye insertion approach for detection of DNA damage. The capture DNA with overhanging 3'-terminus was immobilized on silicon surface to hybridize with target DNA. The intercalation of cyanine dye of thiazole orange (TO) to the double helix structure of DNA (dsDNA) allowed intense enhancement of fluorescence signal. The DNA damage with chemicals led to poor intercalation of TO into double helix structure, resulting in the decrease of the fluorescence signal. This signal decrease could be further enhanced by exonuclease III (Exo III). With this approach, the target DNA could be detected down to 47 fM. Seven chemicals were chosen as models to monitor DNA damage. The results suggested that the present strategy could be developed to detect DNA damage, to classify the damaging mechanism with chemicals and to estimate the toxic effect of chemicals.
authors.affiliation: Key Laboratory of Environmental Medicine Engineering, Ministry of Education, School of Chemistry and Chemical Engineering, Southeast University, Jiangning District 211189, Nanjing, Jiangsu Province, PR China.
authors.forename: Qian
authors.initials: Q
authors.lastname: Lu
authors.forename: Zhenxian
authors.initials: Z
authors.lastname: Zhou
authors.forename: Yuan
authors.initials: Y
authors.lastname: Mei
authors.forename: Wei
authors.initials: W
authors.lastname: Wei
authors.forename: Songqin
authors.initials: S
authors.lastname: Liu
mesh_terms: D020029:Base Pairing; D052160:Benzothiazoles / Q000737:chemistry*; D004247:DNA; D053903:DNA Breaks, Double-Stranded; D015342:DNA Probes / Q000138:chemical synthesis / Q000737:chemistry*; D005090:Exodeoxyribonucleases / Q000737:chemistry*; D005456:Fluorescent Dyes / Q000737:chemistry*; D007364:Intercalating Agents / Q000737:chemistry*; D057230:Limit of Detection; D011804:Quinolines / Q000737:chemistry*; D012825:Silicon / Q000737:chemistry; D012996:Solutions; D013050:Spectrometry, Fluorescence / Q000379:methods
chemical_list: D052160:Benzothiazoles; D015342:DNA Probes; D005456:Fluorescent Dyes; D007364:Intercalating Agents; D011804:Quinolines; D012996:Solutions; C051000:thiazole orange; D004247:DNA; D005090:Exodeoxyribonucleases; C020133:exodeoxyribonuclease III; D012825:Silicon
keywords: Affinity; DNA damage; Exonuclease III; Fluorescence; Intercalation