Division of Intramural Research, Genome Technology Branch, National Human Genome Research Institute, National Institutes of Health, Bethesda, MD 20892-8004, USA.
Nucleic Acids Res. 2014 Apr;42(7):4257-69. doi: 10.1093/nar/gkt1399. Epub 2014 Jan 23.
Retroviruses integrate into the host genome in patterns specific to each virus. Understanding the causes of these patterns can provide insight into viral integration mechanisms, pathology and genome evolution, and is critical to the development of safe gene therapy vectors. We generated murine leukemia virus integrations in human HepG2 and K562 cells and subjected them to second-generation sequencing, using a DNA barcoding technique that allowed us to quantify independent integration events. We characterized >3,700,000 unique integration events in two ENCODE-characterized cell lines. We find that integrations were most highly enriched in a subset of strong enhancers and active promoters. In both cell types, approximately half the integrations were found in <2% of the genome, demonstrating genomic influences even narrower than previously believed. The integration pattern of murine leukemia virus appears to be largely driven by regions that have high enrichment for multiple marks of active chromatin; the combination of histone marks present was sufficient to explain why some strong enhancers were more prone to integration than others. The approach we used is applicable to analyzing the integration pattern of any exogenous element and could be a valuable preclinical screen to evaluate the safety of gene therapy vectors.
逆转录病毒以特定于每种病毒的模式整合到宿主基因组中。了解这些模式的原因可以深入了解病毒整合机制、病理学和基因组进化,对于安全基因治疗载体的开发至关重要。我们在人类 HepG2 和 K562 细胞中生成了鼠白血病病毒整合,并对其进行了第二代测序,使用 DNA 条形码技术使我们能够定量独立的整合事件。我们在两个 ENCODE 特征化的细胞系中鉴定了 >3700000 个独特的整合事件。我们发现整合在一组强增强子和活性启动子中高度富集。在两种细胞类型中,大约一半的整合发生在基因组的 <2%中,这表明基因组的影响甚至比以前认为的更窄。鼠白血病病毒的整合模式似乎主要由富含多种活性染色质标记的区域驱动;存在的组蛋白标记足以解释为什么一些强增强子比其他增强子更容易被整合。我们使用的方法适用于分析任何外源元件的整合模式,并且可以作为一种有价值的临床前筛选,以评估基因治疗载体的安全性。