Simultaneous detection of multiple DNA damage types by multi-colour fluorescent labelling

Herein we present an assay allowing concurrent detection of oxidative DNA damage and photoproducts. We apply DNA repair enzymes specific for each lesion type to incorporate spectrally distinct fluorescent nucleotides, enabling simultaneous quantification of the lesions on individual DNA molecules. W...

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Bibliographic Details
Published in:Chemical communications (Cambridge, England) England), 2019-09, Vol.55 (76), p.11414-11417
Main Authors: Torchinsky, Dmitry, Michaeli, Yael, Gassman, Natalie R, Ebenstein, Yuval
Format: Article
Language:English
Subjects:
Deoxyribonucleic acid
DNA
DNA damage
Fluorescence
Nucleotides
Repair
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Summary:Herein we present an assay allowing concurrent detection of oxidative DNA damage and photoproducts. We apply DNA repair enzymes specific for each lesion type to incorporate spectrally distinct fluorescent nucleotides, enabling simultaneous quantification of the lesions on individual DNA molecules. We follow the repair of both damage types in skin cells exposed to artificial sunlight. Specific and simultaneous quantitation of DNA adducts is a major obstacle. Using repair enzymes, we present a protocol to quantify two types of DNA lesions simultaneously on the same DNA molecule and examine repair dynamics by single-molecule imaging.
ISSN:1359-7345
1364-548X
DOI:10.1039/c9cc05198h