DNA damage response is indispensable for life. As DNA is constantly exposed to various damaging engines, lack of their repair leads to genomes'instability and diseases. We developed a method based on biological property of the RPA2, to visualize single strand DNA tract during DNA repair in different cell cycle stages.
Traditional method for single strand DNA detection utilize BrdU antibodies. This approach can be used for actively replicating cells. Additionally, this antibody tend to cross-react with other nucleotide analogs, limiting their use.
Unlike BRDU, our approach has a higher signal to noise ratio, making it usable in every cycle phase. Understanding the action of different DNA repair pathways in various cell cycle phases and potentially non replicating cells is crucial. Our protocol provides a new approach and tool to visualize single stranded DNA foci, where classical approaches utilizing BrdU are limited.
A comprehensive understanding of DNA repair processes could serve as therapeutic targets to treat cancer, aging, and neurological diseases. Most of our cells are terminally differentiated and therefore do not replicate and divide. It is crucial to gain a better understanding of how these cells repair various DNA lesions.
Therefore, our laboratory will be mainly focusing on post replicative DNA repair processes and developing new tools to study them.
