CometChip enables parallel analysis of multiple DNA repair activities

• Published in: DNA repair 2021-10, Vol.106, p.103176-103176, Article 103176
• Main Authors: Ge, Jing, Ngo, Le P., Kaushal, Simran, Tay, Ian J., Thadhani, Elina, Kay, Jennifer E., Mazzucato, Patrizia, Chow, Danielle N., Fessler, Jessica L., Weingeist, David M., Sobol, Robert W., Samson, Leona D., Floyd, Scott R., Engelward, Bevin P.
• Format: Article
• Language: English
• Subjects: Cell Line; Cell Line, Tumor; Comet assay; Comet Assay - methods; CometChip; DNA - drug effects; DNA - metabolism; DNA - radiation effects; DNA Damage; DNA End-Joining Repair; DNA Repair; High-Throughput Screening Assays - methods; Humans; Mutagens - toxicity
• ISSN: 1568-7864; 1568-7856
• DOI: 10.1016/j.dnarep.2021.103176

•CometChip enables rapid DNA damage analysis in a 96-well format.•CometChip applications include detection of single and double strand breaks.•Defects in BER, NER, and NHEJ can be revealed using the CometChip.•Purified DNA repair enzymes can be added to the CometChip to reveal their substrates.•Results show utility of the CometChip for future molecular epidemiology studies. DNA damage can be cytotoxic and mutagenic, and it is directly linked to aging, cancer, and other diseases. To counteract the deleterious effects of DNA damage, cells have evolved highly conserved DNA repair pathways. Many commonly used DNA repair assays are relatively low throughput and are limited to analysis of one protein or one pathway. Here, we have explored the capacity of the CometChip platform for parallel analysis of multiple DNA repair activities. Taking advantage of the versatility of the traditional comet assay and leveraging micropatterning techniques, the CometChip platform offers increased throughput and sensitivity compared to the traditional comet assay. By exposing cells to DNA damaging agents that create substrates of Base Excision Repair, Nucleotide Excision Repair, and Non-Homologous End Joining, we show that the CometChip is an effective method for assessing repair deficiencies in all three pathways. With these applications of the CometChip platform, we expand the utility of the comet assay for precise, high-throughput, parallel analysis of multiple DNA repair activities.