Novel approaches to map DNA replication traffic in a genome-wide scale

项目摘要 该项目的总体目标是建立新的基因组学和生物信息学方法来模拟 DNA在细胞中的复制模式。众所周知，DNA复制的模式与DNA的结构密切相关。 细胞类型。细胞类型之间的这些差异与细胞分型过程中基因组的完整性有关。 过渡。例如，我们最近发现，DNA复制过程中的损伤在诱导过程中增加。 从一种细胞类型转变为另一种细胞类型。然而，缺乏观察复制模式的方法 在人类细胞中。DNA复制在不同的细胞类型之间在起始位置、复制方向、复制起始时间和复制起始时间上是不同的。 分叉进程，以及启动和完成的时间。目前还没有一种方法可以 全面绘制DNA复制到基因组的进程。因为DNA的重要性 由于在细胞增殖中存在复制，因此需要开发此类方法。为了能够检查 在人类细胞中的DNA聚合酶的进展，我们将开发一种新的方法来映射DNA复制 全基因组，通过在DNA复制过程中掺入核苷酸类似物，并对所得DNA进行测序 分子通过纳米孔测序和记录电信号。与此同时，我们将开发新的 生物信息学的方法来可靠地检查电信号和识别DNA的碱基或区域 复制，用于不同细胞类型之间或健康和患病组织之间的比较。成功 这项技术建立将大大增加我们对复制、遗传稳定性和细胞 增殖，并允许社区表征DNA复制过程中的差异， 细胞类型。

PROJECT SUMMARY The overarching goal of the project is to establish novel genomics and bioinformatics approaches to model patterns of DNA replication in cells. It is known that the patterns of DNA replication are intimately linked to the cell type. These differences between cell types are relevant to the integrity of the genome during cell type transitions. For example, we recently found that damage during DNA replication is increased during the induced transition from one cell type to another. However, there is a lack of methodology to observe replication patterns in human cells. DNA replication differs between different cell types in the location of initiation, the direction of fork progression, and the timing of initiation and completion. There is currently no method that can comprehensively map the progression of DNA replication to the genome. Because of the importance of DNA replication in cell proliferation, there is need for the development of such methods. To be able to examine the progression of DNA polymerases in human cells, we will develop a novel methodology to map DNA replication genome-wide, by incorporating nucleotide analogs during DNA replication, and sequencing the resulting DNA molecules by Nanopore sequencing and recording the electrical signals. In parallel, we will develop novel bioinformatics approaches to reliably examine the electrical signals and identify bases or regions of DNA replication, for comparison between different cell types, or between healthy and diseased tissues. Successful establishment of this technology will greatly increase our knowledge of replication, genetic stability and cell proliferation, and allow the community to characterize differences in the progression of DNA replication between cell types.