用于全基因组绘制含8-氧代-2'-脱氧鸟苷的受损DNA图谱的氧化DNA免疫沉淀测序法（OxiDIP-Seq）
OxiDIP-Seq for Genome-wide Mapping of Damaged DNA Containing 8-Oxo-2'-Deoxyguanosine.
作者信息
Gorini Francesca, Scala Giovanni, Ambrosio Susanna, Majello Barbara, Amente Stefano
机构信息
Department of Molecular Medicine and Medical Biotechnologies, University of Naples 'Federico II', Naples, Italy.
Department of Biology, University of Naples 'Federico II', Naples, Italy.
出版信息
Bio Protoc. 2022 Nov 5;12(21). doi: 10.21769/BioProtoc.4540.
8-oxo-7,8-dihydro-2'-deoxyguanosine (8-oxodG) is considered to be a premutagenic DNA lesion generated by 2'-deoxyguanosine (dG) oxidation due to reactive oxygen species (ROS). In recent years, the 8-oxodG distribution in human, mouse, and yeast genomes has been underlined using various next-generation sequencing (NGS)-based strategies. The present study reports the OxiDIP-Seq protocol, which combines specific 8-oxodG immuno-precipitation of single-stranded DNA with NGS, and the pipeline analysis that allows the genome-wide 8-oxodG distribution in mammalian cells. The development of this OxiDIP-Seq method increases knowledge on the oxidative DNA damage/repair field, providing a high-resolution map of 8-oxodG in human cells.
8-氧代-7,8-二氢-2'-脱氧鸟苷（8-oxodG）被认为是由活性氧（ROS）导致的2'-脱氧鸟苷（dG）氧化产生的一种前诱变DNA损伤。近年来，利用各种基于新一代测序（NGS）的策略已经突显了人类、小鼠和酵母基因组中8-oxodG的分布情况。本研究报告了OxiDIP-Seq方案，该方案将单链DNA的特异性8-oxodG免疫沉淀与NGS相结合，以及允许在哺乳动物细胞中进行全基因组8-oxodG分布分析的流程。这种OxiDIP-Seq方法的开发增加了对氧化性DNA损伤/修复领域的认识，提供了人类细胞中8-oxodG的高分辨率图谱。
Zotero 插件