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Fluorescent Hairpin Probes for the Detection of Chemically-Induced DNA Damage Open Access


Other title
Hairpin probe
DNA damage
Molecular beacon
Type of item
Degree grantor
University of Alberta
Author or creator
Shire, Zahra J
Supervisor and department
Loppnow, Glen ( Department of Chemistry)
Examining committee member and department
Gallin, Warren ( Department of Biological Sciences)
Campbell, Robert ( Department of Chemistry)
Department of Chemistry

Date accepted
Graduation date
Master of Science
Degree level
In this thesis, we develop a sensitive, robust, accurate method to detect chemically-induced DNA damage by three therapeutic agents, cisplatin, psoralen and busulfan. The method uses fluorescence spectroscopy and fluorescent hairpin or molecular beacon probes in a hybridization assay for chemically-induced ssDNA damage probed in solution and on a microarray platform. Damage is confirmed by matrix-assisted laser desorption / ionization time-of-flight mass spectrometry (MALDI-TOF MS) and UV absorbance spectroscopy. The decrease in fluorescence upon damage scales with the number of mutation sites, drug doses and irradiation time. All these results indicate that the hybridization probes can quantitatively and selectively detect the number of DNA lesions/strand for chemically-induced DNA damage. Finally, fluorescent hairpin probes have some limitations, particularly with regards to their modifications and costs. However, the development of easy, fast and reliable assays is essential for biological as well as clinical applications, but requires these limitations to be addressed.
Permission is hereby granted to the University of Alberta Libraries to reproduce single copies of this thesis and to lend or sell such copies for private, scholarly or scientific research purposes only. Where the thesis is converted to, or otherwise made available in digital form, the University of Alberta will advise potential users of the thesis of these terms. The author reserves all other publication and other rights in association with the copyright in the thesis and, except as herein before provided, neither the thesis nor any substantial portion thereof may be printed or otherwise reproduced in any material form whatsoever without the author's prior written permission.
Citation for previous publication
Shire, Z. J.; Loppnow, G. R. Anal. Bioanal. Chem., 2012, 403, 179 -184Shire, Z. J.; Loppnow, G. R. Photochem. Photobiol. 2012, DOI: 1111/j.1751-1097.2012.01109.x

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