生命学院学术报告

题目：DNA Damage and Repair Map of the Entire Human Genome

报告人：Jinchuan Hu

Post-Doctor Fellow

Department of Biochemistry and Biophysics

University of North Carolina School of Medicine, Chapel Hill

时间：2015年11月24日（星期二）13：00-14：00

地点：208会议室

联系人：伊成器

摘要：

DNA base lesions are caused by various genotoxic agents, including UV radiation and cisplatin. These DNA lesions can interfere with DNA replication and transcription, lead to mutation and cell death, and finally cause cancer and other disease. We developed methods for detecting DNA damage (Damage-seq) and repair (XR-seq) sites at single nucleotide resolution and have generated damage and repair maps for the entire human genome. In Damage-seq, the exact locations of DNA damage are determined by the arrested DNA polymerase at damage sites. We generated the damage maps of CPDs, (6-4)PPs and cisplatin-adducts in human skin fibroblasts and lymphocytes. The results showed that di-pyrimidines (TT/TC) and GG are the dominant modified nucleotides for UV and cisplatin induced DNA damages, respectively, validating the specificity of this method. In XR-seq, the excised oligonucleotides containing DNA damage were captured by TFIIH-IP and subjected to high-throughput sequencing. By using cell lines defective in either transcription-coupled repair or global repair, we generated (6-4)PP and CPD repair maps of both pathways. Both methods can be used to study the association of DNA damage and repair with transcription, chromatin states and other factors. We can also compare the damage map by Damage-seq and repair map by XR-seq in the same cell line under the same condition. These methods can be extended to more damage types and have the potential use in cancer prevention and chemotherapy.

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