DNA Repair Capacity Kit

Technology   

With this novel DNA repair capability assessment kit, cells are transfected with a library of plasmid DNA reporters containing various types or doses of DNA damage. The repair of DNA damage results in a measurable change in fluorescent reporter protein expression. Therefore, each fluorescent reporter color corresponds to the repair of a different type or dose of DNA damage. Fluorescence is measured by flow cytometry or laser scanning cytometry. A 96 well plate of cell samples can be assessed at once, allowing for high throughput and straightforward multiplexing.  

Problem Addressed    

In a human cell, normal metabolic activities and environmental factors such as radiation can cause as many as one million individual molecular lesions per day. To maintain genetic integrity and normal cell functions, a mechanism called DNA damage repair is crucial in allowing a cell to identify DNA damage and mediate its repair. DNA repair capacities differ between species and individuals within species. Quantifying this difference is important for research purposes and for clinical diagnostics (e.g. personalized disease prevention, prescribing chemotherapy tailored to an individual patient based on differences in DNA repair capacity of the patient). Furthermore, the different types of DNA damage necessitate different molecular pathways of DNA repair.  

Advantages

• Targets several different pathways in a single assay
•  Offers faster and easier tests than current methods that only assess a single type of damage, by measuring repair of four or more types of damage simultaneously
• Enables users to automate DNA repair tests using robotics, saving time compared to current tests that require laborious manual procedures
• Achieves high fidelity through rigorous controls  

Publications

"Multiplexed DNA Repair Assays for Multiple Lesions and Multiple Doses Via Transcription Inhibition and Transcriptional Mutagenesis." Proceedings of the National Academy of Sciences of the United States of America 111, no. 18 (May 6, 2014): E1823-E1832.