Year: 2013
Type: article
Abstract: DNA damage due to reactive oxygen species (ROS) has severe consequences to cellular metabolism and its accumulation can lead to many diseases. One type of ROS-induced DNA damage is produced primarily by hydroxyl radicals which generate both single- and double-strand breaks in DNA. Our aim is to map the landscape of such damage on genome-wide scale at high resolution. A genome-wide map of ROS-induced DNA strand breaks is likely to show a greater amount of damage in open chromatin due to higher accessibility of genomic DNA in these regions. In order to map oxidative damage throughout a genome at single-nucleotide resolution, we have developed a new method (OH-Seq) to biochemically process oxidatively-damaged genomic DNA to make it suitable for high-throughput sequencing. We use gamma radiation to generate hydroxyl radicals in vivo and then process the damaged genomic DNA to tag the breaks with a sequencing adapter. By using Illumina high-throughput sequencing to locate such tagged sites in the genome, it is possible to obtain a high resolution genome-wide map of DNA strand breaks. This map yields a detailed picture of functional regions and transcription factor binding sites in the genome, and has the potential to provide a new level of detail in the study of DNA repair and oxidatively-induced aging.
Authors: Hari K.K. Subramanian, Sarah E. Bernard, Cheryl Chiang, Stephen C. J. Parker, Thomas D. Tullius
Language: English
Topic: DNA Repair Mechanisms
Subfield: Molecular Biology
Domain: Life Sciences
Open Access status: bronze
OpenAlex ID: W2020954463