Genome wide identification of priming sites for Okazaki fragments

冈崎片段引发位点的全基因组鉴定

基本信息

批准号：
BB/K018272/1
负责人：
Nigel Burroughs
金额：
$ 89.93万
依托单位：
University of Warwick
依托单位国家：
英国
项目类别：
Research Grant
财政年份：
2013
资助国家：
英国
起止时间：
2013 至无数据
项目状态：
已结题

来源：
https://gtr.ukri.org/projects?ref=BB%2FK018272%2F1
关键词：
Genome wide identification priming sites

项目摘要

Our genetic blueprint is encoded by chromosomal molecules that consist of DNA as well as chromatin. The chromatin determines expression states of individual genes. As we go through life, cells get damaged and die. Thus, tissues are constantly regenerating through cell divisions. Every time a cell divides, the DNA and the associated chromatin have to be duplicated. This happens by the process of DNA replication. DNA replication is initiated at specific sites in the genome called origins, and progresses in a bidirectional manner from these. Since DNA consists of two anti-parallel strands, and these two strands only can be copied in one direction, one of the strands, the leading, is synthesized in a continuous manner, while the other, the lagging, is synthesized as fragments that are subsequently put together to form the final product. Each time the DNA replication is initiated on the lagging strand, a priming event takes place, but virtually nothing is known about what determines where these primers are put down. We have developed a novel method for determining the position of the priming sites on the genomic level using Next Generation Sequencing. Using this technical breakthrough, we will answer fundamental questions about the replication process that would not have been possible to answer before. We will be able to identify positions in the genome where the replication process slows down or terminates, as at these positions priming is site-specific. We also will be able to identify region were there are "problems" during the replication process due to infrequent or too frequent priming events. Finally, we will investigate whether chromatin affects where primers are put down. The project will answer some very fundamental questions about the replication process underlying all cellular life, and will increase our knowledge about what can go wrong during this essential process, thus giving us insight into the genetic instability that underlies cancer and ageing.

我们的遗传蓝图由由 DNA 和染色质组成的染色体分子编码。染色质决定单个基因的表达状态。在我们的一生中，细胞会受损并死亡。因此，组织通过细胞分裂不断再生。每次细胞分裂时，DNA 和相关染色质都必须复制。这是通过 DNA 复制过程发生的。 DNA 复制在基因组中称为起点的特定位点开始，并从这些位点开始以双向方式进行。由于DNA由两条反向平行链组成，并且这两条链只能沿一个方向复制，因此其中一条链（前导链）以连续方式合成，而另一条链（后链）以片段形式合成，随后组合在一起形成最终产品。每次 DNA 复制在滞后链上启动时，都会发生启动事件，但实际上我们对决定这些引物放置位置的因素一无所知。我们开发了一种使用下一代测序在基因组水平上确定启动位点位置的新方法。利用这一技术突破，我们将回答以前无法回答的有关复制过程的基本问题。我们将能够识别基因组中复制过程减慢或终止的位置，因为在这些位置上启动是位点特异性的。我们还能够识别复制过程中由于启动事件不频繁或过于频繁而出现“问题”的区域。最后，我们将研究染色质是否影响引物的放置位置。该项目将回答一些关于所有细胞生命的复制过程的非常基本的问题，并将增加我们对这一重要过程中可能出现问题的了解，从而使我们深入了解癌症和衰老背后的遗传不稳定性。