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Single-cell microarray enables high-throughput evaluation of DNA double-strand breaks and DNA repair inhibitors.

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Date
2013-03-15
Authors
Weingeist, David M
Ge, Jing
Wood, David K
Mutamba, James T
Huang, Qiuying
Rowland, Elizabeth A
Yaffe, Michael B
Floyd, Scott
Engelward, Bevin P
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(9 total)
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Abstract
A key modality of non-surgical cancer management is DNA damaging therapy that causes DNA double-strand breaks that are preferentially toxic to rapidly dividing cancer cells. Double-strand break repair capacity is recognized as an important mechanism in drug resistance and is therefore a potential target for adjuvant chemotherapy. Additionally, spontaneous and environmentally induced DSBs are known to promote cancer, making DSB evaluation important as a tool in epidemiology, clinical evaluation and in the development of novel pharmaceuticals. Currently available assays to detect double-strand breaks are limited in throughput and specificity and offer minimal information concerning the kinetics of repair. Here, we present the CometChip, a 96-well platform that enables assessment of double-strand break levels and repair capacity of multiple cell types and conditions in parallel and integrates with standard high-throughput screening and analysis technologies. We demonstrate the ability to detect multiple genetic deficiencies in double-strand break repair and evaluate a set of clinically relevant chemical inhibitors of one of the major double-strand break repair pathways, non-homologous end-joining. While other high-throughput repair assays measure residual damage or indirect markers of damage, the CometChip detects physical double-strand breaks, providing direct measurement of damage induction and repair capacity, which may be useful in developing and implementing treatment strategies with reduced side effects.
Type
Journal article
Subject
DNA double-strand breaks
DNA repair
DNA-PK inhibitors
high throughput
microarray
neutral comet assay
neutral single-cell electrophoresis assay
non-homologous end-joining
Animals
CHO Cells
Cell Line
Chromones
Cricetinae
DNA Breaks, Double-Stranded
DNA Damage
DNA Repair
DNA-Activated Protein Kinase
Drug Resistance, Neoplasm
Enzyme Inhibitors
High-Throughput Screening Assays
Humans
Morpholines
Neoplasms
Permalink
https://hdl.handle.net/10161/15938
Published Version (Please cite this version)
10.4161/cc.23880
Publication Info
Weingeist, David M; Ge, Jing; Wood, David K; Mutamba, James T; Huang, Qiuying; Rowland, Elizabeth A; ... Engelward, Bevin P (2013). Single-cell microarray enables high-throughput evaluation of DNA double-strand breaks and DNA repair inhibitors. Cell Cycle, 12(6). pp. 907-915. 10.4161/cc.23880. Retrieved from https://hdl.handle.net/10161/15938.
This is constructed from limited available data and may be imprecise. To cite this article, please review & use the official citation provided by the journal.
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Scholars@Duke

Floyd

Scott Richard Floyd

Gary Hock and Lyn Proctor Associate Professor of Radiation Oncology
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