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

TitleSingle-cell microarray enables high-throughput evaluation of DNA double-strand breaks and DNA repair inhibitors.
Publication TypeJournal Article
Year of Publication2013
AuthorsWeingeist, DM, Ge, J, Wood, DK, Mutamba, JT, Huang, Q, Rowland, EA, Yaffe, MB, Floyd, S, Engelward, BP
JournalCell Cycle
Date Published2013 Mar 15
KeywordsAnimals, Cell Line, CHO Cells, 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

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.

Alternate JournalCell Cycle
PubMed ID23422001
PubMed Central IDPMC3637349
Grant ListCA112967 / CA / NCI NIH HHS / United States
ES015339 / ES / NIEHS NIH HHS / United States
P30-ES002109 / ES / NIEHS NIH HHS / United States
R21-ES019498 / ES / NIEHS NIH HHS / United States
T32-ES007020 / ES / NIEHS NIH HHS / United States
U01-ES016045 / ES / NIEHS NIH HHS / United States