Approach for assessing total cellular DNA damage.
The capability to relate phenotypic effects to damage associated with either the mitochondrial or nuclear genome is especially useful under a number of circumstances. Potential hazardous exposures can be evaluated for genotoxicity and related to diseases, particularly cancer. The correlation of DNA damage with adverse health effects is also important in evaluating the safety of various chemical agents and prospective therapeutics. Many techniques exist that afford the ability to identify and measure cellular DNA damage upon exposure to a suspected genotoxic agent; however, quite often these techniques are limited either by the advanced instrumentation and skill needed to perform the analyses or the amount of time needed and limited information obtained regarding the types of DNA damage generated. Recent advances in cellular-based methods have resulted in the timely and straightforward collection of reliable and specific data regarding levels of damage and the identity of the damage products. Antibodies developed for DNA damage lesions allow for the direct measurement of those lesions within a population of exposed cells, while the automation of the single-cell gel electrophoresis (comet) assay and the use of scoring software have led to rapid and standardized data collection. This essay describes the usefulness of these approaches, while providing a brief experimental overview of the techniques.
Roy
Marc D
MD
Polymers Division, Biomaterials Group, National Institute of Standards and Technology, Gaithersburg, MD, USA.
D020552:Comet Assay; D004249:DNA Damage; D004272:DNA, Mitochondrial; D019084:Fluorescent Antibody Technique, Indirect; D006151:Guanosine / Q000031:analogs & derivatives / Q000032:analysis; D009153:Mutagens / Q000032:analysis*
D004272:DNA, Mitochondrial; D009153:Mutagens; D006151:Guanosine; C046215:8-hydroxyguanosine