Sequencing the Mouse Genome for the Oxidatively Modified Base 8-Oxo-7,8-dihydroguanine by OG-Seq.
Sequencing the Mouse Genome for the Oxidatively Modified Base 8-Oxo-7,8-dihydroguanine by OG-Seq.
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DOI：
10.1021/jacs.6b12604
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发表时间：
2017-02-22
影响因子：
15
通讯作者：
Burrows CJ
中科院分区：
文献类型：
--
作者：
Ding Y;Fleming AM;Burrows CJ
对基因组的氧化损伤可以产生碱基8-氧代-7，8-二氢鸟嘌呤（OG）。体外研究表明，暴露于活性氧后，OG优先在5′-GG-3′序列背景下形成。在此，通过开发“OG-Seq”来研究基因组中的OG位置，以通过下一代测序以0.15-kb分辨率对OG位点进行测序。本研究的结果发现，野生型小鼠胚胎成纤维细胞中有10000个区域的OG富集，当OG修复糖基化酶Ogg 1被敲除时，有18000个区域的OG富集。如果基因启动子和UTR位点随机分布在整个基因组中并与反应性5′-GG-3′序列相关，则这些位点比预期的具有更多的OG富集位点，这一结果支持了数十年的体外研究。OG的测序为解决围绕这种修饰的DNA碱基的化学和生物学问题铺平了道路，例如它在疾病特异性突变中的作用及其在基因调控中的表观遗传潜力。
Oxidative damage to the genome can yield the base 8-oxo-7,8-dihydroguanine (OG). In vitro studies suggested OG would preferentially form in 5′-GG-3′ sequence contexts after exposure to reactive oxygen species. Herein, OG locations in the genome were studied by development of “OG-Seq” to sequence OG sites via next-generation sequencing at ∼0.15-kb resolution. The results of this study found ∼10000 regions of OG enrichment in WT mouse embryonic fibroblasts and ∼18000 regions when the OG repair glycosylase Ogg1 was knocked out. Gene promoters and UTRs harbor more OG-enriched sites than expected if the sites were randomly distributed throughout the genome and correlate with reactive 5′-GG-3′ sequences, a result supporting decades of in vitro studies. Sequencing of OG paves the way to address chemical and biological questions surrounding this modified DNA base, such as its role in disease-specific mutations and its epigenetic potential in gene regulation.