4.3 Article

CometChip enables parallel analysis of multiple DNA repair activities

期刊

DNA REPAIR
卷 106, 期 -, 页码 -

出版社

ELSEVIER
DOI: 10.1016/j.dnarep.2021.103176

关键词

DNA damage; DNA repair; Comet assay; CometChip

资金

  1. National Institute of Health (NIH) National Cancer Institute [R01 ES022872]
  2. National Institute of Environmental Health Sciences Superfund Basic Research Program [P42 ES027707]
  3. NIH [R01 CA148629, GM087798, ES02116]
  4. MIT Center for Environmental Health Sciences [P30-ES002109]
  5. Burroughs Wellcome Fund Career Award for Medical Scientists [1010922]

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DNA damage is directly linked to aging, cancer, and other diseases, and cells have evolved highly conserved DNA repair pathways to counteract its deleterious effects. The CometChip platform offers increased throughput and sensitivity for parallel analysis of multiple DNA repair activities, expanding the utility of the comet assay for precise, high-throughput assessment of repair deficiencies in different pathways.
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 micro patterning 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.

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