Sensitive CometChip assay for screening potentially carcinogenic DNA adducts by trapping DNA repair intermediates
Name
gkz1077.pdf
Description
Published version
Size
1.35 MB
Format
Adobe PDF
Checksum (MD5)
2edcce0d565e3cc35b3f61526ad0e485
Author(s)
Ngo, Le P
Owiti, Norah A
Su, Yang
Ge, Jing
Han, Jongyoon
Samson, Leona D
Engelward, Bevin P
Date Issued
December 11, 2019
Journal
Nucleic Acids Research
Publisher
Oxford University Press (OUP)
Citation
Ngo, Le P. et al. “Sensitive CometChip assay for screening potentially carcinogenic DNA adducts by trapping DNA repair intermediates.” Nucleic Acids Research 48 (2020): e13 © 2020 The Author(s)
Version
Final published version
Abstract
Genotoxicity testing is critical for predicting adverse effects of pharmaceutical, industrial, and environmental chemicals. The alkaline comet assayis an established method for detecting DNA strandbreaks, however, the assay does not detect potentially carcinogenic bulky adducts that can arise whenmetabolic enzymes convert pro-carcinogens into ahighly DNA reactive products. To overcome this, weuse DNA synthesis inhibitors (hydroxyurea and 1-β-d-arabinofuranosyl cytosine) to trap single strandbreaks that are formed during nucleotide excision repair, which primarily removes bulky lesions. In thisway, comet-undetectable bulky lesions are convertedinto comet-detectable single strand breaks. Moreover, we use HepaRG™cells to recapitulatein vivometabolic capacity, and leverage the CometChip platform (a higher throughput more sensitive comet as-say) to create the ‘HepaCometChip’, enabling the detection of bulky genotoxic lesions that are missed bycurrent genotoxicity screens. The HepaCometChip thus provides a broadly effective approach for detection of bulky DNA adducts.
Subjects
Genetics
MIT Department
Massachusetts Institute of Technology. Department of Biological Engineering
Massachusetts Institute of Technology. Department of Biology
Massachusetts Institute of Technology. Department of Electrical Engineering and Computer Science
Terms of Use
Creative Commons Attribution 4.0 International license
Persistent DSpace Link
DOI of Published Version
https://doi.org/10.1093/nar/gkz1077
