Advantages:
- The modified comet assay delivers consistent, reproducible results, ensuring reliable data for bacterial pathogenesis and DNA damage/repair studies
- A cost-effective and user-friendly antimicrobial peptide, making the modified comet assay an economical solution
- The method is versatile, applicable to bacteria, mammalian cells, microbiome, and disease research
Summary:
Chronic exposure to genotoxic agents, including bacterial infections, plays a significant role in the development of chronic diseases and cancer. Bacterial infections, particularly Mycoplasma and Staphylococcus aureus, are increasingly recognized for their ability to induce DNA damage yet studying these effects in vitro has been challenging. Traditional DNA damage assessment methods, such as the comet assay, are hindered by bacterial interference, particularly from bacterial DNA and peptidoglycan structures, which create background fluorescence and disrupt accurate scoring of DNA damage in mammalian cells. This issue limits the ability to study bacterially induced DNA damage effectively and to understand the mechanisms contributing to genomic instability and disease pathogenesis.
Our researchers have developed an innovative antibacterial-modified comet assay that provides a groundbreaking solution for studying DNA damage in mammalian cells co-cultured with bacteria, specifically addressing the challenge of bacterial interference. By utilizing lysostaphin, a targeted antimicrobial peptide, bacterial background staining is effectively removed, ensuring clear, reproducible results without inducing artefactual DNA damage in S. aureus co-cultures. This advanced approach enables precise measurement of oxidative stress-induced DNA damage, crucial for understanding bacterial infections like S. aureus and Mycoplasma, which can disrupt genomic stability and DNA repair. The assay is highly versatile, offering a reliable method for exploring bacterial pathogenesis, microbiome-related diseases, and potential therapeutic strategies.
Lysostaphin treatment during the comet assay removes background staining interference from Staphylococcus aureus, allowing the scoring of DNA damage in eukaryotic cells. (A) Illustration of the background staining caused by S. aureus in the alkaline comet assay, following infection of human keratinocytes cells; (B) Representative image of a comet from (A), under analysis by Comet IV scoring software, showing that scoring is impossible due to the interference from the background staining; (C) Representative comet assay image illustrating that treatment with 1 mg/mL lysostaphin removes the interfering background staining caused by S. aureus infected human keratinocytes; (D) Illustration of successful scoring of the same cells as in (C), by Comet IV software.
Desired Partnerships:
- License
- Sponsored Research
- Co-Development