The detection of DNA strand breaks and repair is based on progressive DNA unwinding under highly controlled conditions of alkaline pH, time and temperature. DNA “open sites” are the starting points for the unwinding process. They occur naturally during cell cycle processes at replication forks or the ends of chromosomes. As yet, they may additionally be induced by reactive oxygen species, irradiation or genotoxic chemicals. As basis of the gTOXXs Analyzer, the FADU assay is used which is based on a sensitive fluorescent probe which serves as marker for the loss of intact double stranded DNA. (Decrease in the fluorescence intensity indicates an increase in DNA unwinding and consequently a greater number of DNA strand breaks). The gTOXXs Analyzer provides an alternative to animal testing!
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Methods & Expertise for Research Infrastructure
The gTOXXs Analyzer speeds up the risk assessment of test substances, as well as the DNA repair capacity in various human cells lines, by use of the AUREA assay (the automated FADU assay). The automation of DNA strand break detection enables reliable determination of the genotoxic potential of chemical, nanoparticle or biological substances. The gTOXXs operation time, reproducibility and workflow routine is clearly superior to other established genotoxicity assays, like the COMET assay. DNA strand breaks are reliably detected even after short cell exposure (≥10 min) to well-known genotoxic compounds at low concentrations (Moreno-Villanueva et al, 2011). The rapid assessment of the genotoxic potency significantly enhances product safety testing in scientific as well as commercial large-scale projects. With the validation of the gTOXXs for 3D skin models completely new applications become available.
• DNA damage detection - Dose-response relationship
• DNA repair after defined induction of damage
• DNA interstrand crosslinks
• Oxidative DNA lesions
• Post-traumatic stress disorder (PTSD)
• Individualized cancer therapy (chemotherapy and radiation therapy)