An Experimental Model for Assessment of Global DNA Repair Capacity

Abstract

Cellular DNA is constantly subjected to a significant amount of damage resulting from extracellular as well as intracellular processes. There are various mechanisms to repair DNA damage among which nucleotide excision repair (NER) pathway has the highest degree of versatility for recognizing and repairing potentially hazardous DNA modifications. It is difficult, however, to measure the rate of DNA repair as the synthesis of DNA related to repair constitutes only a small fraction of the overall rate of DNA synthesis in the cell. The current paper presents an experimental setup that allows measuring the rate of repair DNA synthesis based on suppression of the replicative DNA synthesis in order to differentiate the repair-associated synthetic activity. The proposed methodology allows for assessment of repair rate of the DNA damage of any type and in any DNA region undergoing repair irrelevant of the structure of the region in question. It could serve as a basis for development of diagnostic methods for assessment of the capacity for global DNA repair.