Single molecule analysis of genome replication

基因组复制的单分子分析

基本信息

批准号：
BB/N016858/1
负责人：
Conrad Nieduszynski
金额：
$ 87.82万
依托单位：
University of Oxford
依托单位国家：
英国
项目类别：
Research Grant
财政年份：
2016
资助国家：
英国
起止时间：
2016 至无数据
项目状态：
已结题

来源：
https://gtr.ukri.org/projects?ref=BB%2FN016858%2F1
关键词：
Single molecule analysis genome replication

项目摘要

All cells contain a complete copy of the organism's DNA, the genetic blueprint of life, packaged into discrete units called chromosomes. Since new cells need a copy of the genetic material, the chromosomes must be completely and accurately replicated before the cell can divide. Our research aims to determine how cells ensure that the replication of each chromosome is completed before cell division. Our experimental data have revealed the average pattern of chromosome replication in populations of millions of cells - that is the regions that are replicated first, second and so on. However, in preliminary data we have discovered that individual cells do not replicate their chromosomes with this 'average' pattern; the pattern of DNA replication in single cells is hidden by the population average. Consequently rare problems during DNA replication, including delays in completing replication, cannot currently be readily detected. Therefore, we are developing novel methods to directly measure DNA replication in single molecules. In one approach, we are using a transformative single-molecule sequencing technology to detect the pattern of DNA replication in thousands of long molecules. In addition, we will make use of recent advances in imaging technologies to directly visualise DNA replication in whole chromosomes. These complementary approaches will, for the first time, reveal the pattern of DNA replication on individual chromosomes. This is important because a single DNA replication error on one chromosome in a single cell division can give rise to genomic disorders, including cancer.

所有细胞都包含生物体 DNA（生命的遗传蓝图）的完整副本，被包装成称为染色体的离散单元。由于新细胞需要遗传物质的副本，因此在细胞分裂之前染色体必须完整且准确地复制。我们的研究旨在确定细胞如何确保在细胞分裂之前完成每条染色体的复制。我们的实验数据揭示了数百万个细胞群中染色体复制的平均模式，即首先复制的区域、第二个复制的区域等。然而，在初步数据中，我们发现单个细胞并不以这种“平均”模式复制其染色体。单细胞中 DNA 复制的模式被群体平均值所掩盖。因此，DNA 复制过程中的罕见问题，包括完成复制的延迟，目前还无法轻易检测到。因此，我们正在开发直接测量单分子 DNA 复制的新方法。在一种方法中，我们使用革命性的单分子测序技术来检测数千个长分子中的 DNA 复制模式。此外，我们将利用成像技术的最新进展，直接可视化整个染色体中的 DNA 复制。这些互补的方法将首次揭示单个染色体上 DNA 复制的模式。这一点很重要，因为单次细胞分裂中一条染色体上的单个 DNA 复制错误可能会导致基因组疾病，包括癌症。