Single molecule analysis of Human DNA replication

人类 DNA 复制的单分子分析

• 批准号: BB/Y00549X/1
• 负责人: Conrad Nieduszynski
• 金额: $ 82.13万
• 依托单位: Earlham Institute
• 依托单位国家: 英国
• 项目类别: Research Grant
• 财政年份: 2024
• 资助国家: 英国
• 起止时间: 2024 至 无数据
• 项目状态: 未结题
• 来源: https://gtr.ukri.org/projects?ref=BB%2FY00549X%2F1
• 关键词: Single; molecule; analysis; Human; DNA

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. This requires the process of DNA replication to start at thousands of sites across the chromosomes - called DNA replication initiation sites. Our project aims to determine the location of replication initiation sites in human cells. This is important because the location and distribution of replication initiation sites have been implicated in causing human diseases such as cancer.It has been challenging to identify DNA replication initiation sites in human cells, because there is a lot of variability between cells. To date, most experiments have used the average from millions of cells, but this hides the variability between cells. We have developed a novel technology that identifies DNA replication initiation sites on thousands of single molecules that each originated in a single cell. Our approach can also identify specific sequences that are challenging to copy. This will allow us to test the hypothesis that some sites, thought to be involved in replication initiation, in fact are sites that impede DNA replication. Distinguishing between sites that initiate versus impede DNA replication will be crucial in understanding the causes of genetic instability that underlie some human diseases.

所有细胞均包含生物体DNA的完整副本，即生命的遗传蓝图，被包装成被称为染色体的离散单元。由于新细胞需要遗传物质的副本，因此必须在细胞分裂之前完全准确地复制染色体。这需要DNA复制的过程才能从跨染色体的数千个位点开始 - 称为DNA复制起始位点。我们的项目旨在确定重复起始位点在人类细胞中的位置。这很重要，因为复制起始位点的位置和分布与引起人类疾病（例如癌症）有关。识别人类细胞中DNA复制起始位点的挑战，因为细胞之间存在很多差异。迄今为止，大多数实验都使用了数百万个细胞的平均值，但这隐藏了细胞之间的变异性。我们开发了一种新型技术，该技术可以识别数千个产生单个细胞的单个分子上的DNA复制起始位点。我们的方法还可以确定复制具有挑战性的特定序列。这将使我们能够检验以下假设：某些位点被认为参与复制启动的位置实际上是阻碍DNA复制的站点。区分引发与阻碍DNA复制的部位对于理解某些人类疾病的基因不稳定的原因至关重要。