Base-resolution analysis of cisplatin-DNA adducts at the genome scale

Cisplatin, one of the most widely used anticancer drugs, crosslinks DNA and ultimately induces cell death. However, the genomic pattern of cisplatin-DNA adducts remains unknown, due to the lack of a reliable and sensitive genome-wide method. Here we present “cisplatin-seq” to identify genome-wide cisplatin crosslinking sites at base-resolution. Cisplatin-seq reveals that mitochondrial DNA is a preferred target of cisplatin. For nuclear genome, cisplatin-DNA adducts are enriched within promoters and regions harboring transcription termination sites. While the density of GG dinucleotide determines the initial crosslinking of cisplatin, binding of proteins to the genome largely contributes to the accumulative pattern of cisplatin-DNA adducts. Overall design: Developing a method "Cisplatin-seq" to profile cisplatin-crosslinking site in human genome, using cells treated with cisplatin for 3h, 12h, 24h respectively.

顺铂（cisplatin）是目前临床应用最为广泛的抗癌药物之一，可通过交联DNA最终诱导细胞死亡。然而，由于缺乏可靠且灵敏的全基因组检测方法，顺铂-DNA加合物的全基因组分布模式仍未明确。本文报道了"顺铂-seq（cisplatin-seq）"技术，可在碱基分辨率水平上鉴定全基因组范围内的顺铂交联位点。顺铂-seq研究结果显示，线粒体DNA是顺铂的优先作用靶点。对于核基因组而言，顺铂-DNA加合物在启动子区域及携带转录终止位点的区域显著富集。尽管GG二核苷酸的密度决定了顺铂的初始交联位点，但基因组上蛋白质的结合对顺铂-DNA加合物的累积分布模式起到了主要贡献作用。总体实验设计：本研究开发"顺铂-seq"技术以表征人类基因组中的顺铂交联位点，分别采用经顺铂处理3小时、12小时、24小时的细胞作为实验样本。