Genomic mapping of radiation damage using a next generation sequencer

2015 Fiscal Year Final Research Report

• Project/Area Number: 26550030
• Research Category: Grant-in-Aid for Challenging Exploratory Research
• Allocation Type: Multi-year Fund
• Research Field: Risk sciences of radiation and chemicals
• Research Institution: Hiroshima University
• Principal Investigator: Hiroshi IDE 広島大学, 理学(系)研究科(研究院), 教授 (30223126)
• Project Period (FY): 2014-04-01 – 2016-03-31
• Keywords: 放射線 / DNA損傷

Outline of Final Research Achievements

DNA-protein cross-links (DPCs) are major DNA lesions induced by ionizing radiation and anticancer drugs. However, it remains elusive whether some regions of the genome are more vulnerable with respect to DPC formation than others when cells are exposed to DNA damaging agents. In the present study, we developed a method for genomic mapping of radiation-induced DPCs. Mouse SCCVII tumors were irradiated with X-rays and DNA was purified. Proteins present in DPCs were labeled with biotin and DNA fragments containing DPCs (DPC-DNA) were specifically pulled down using avidin-beads. A synthetic adaptor was ligated at both ends of DPC-DNA. The resulting DNA was subjected to PCR. Analysis of reaction products with agarose gel electrophoresis showed that DPC-DNA was successfully amplified by PCR.

Free Research Field

放射線生物学