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和染色质质组成的染色体分子编码。染色质决定了单个基因的表达态。当我们经历生命时，细胞会损坏并死亡。因此，组织不断地通过细胞分裂再生。每当细胞分裂时，必须重复DNA和相关的染色质。这是通过DNA复制过程发生的。 DNA复制是在称为起源的基因组中的特定位点启动的，并以双向方式进行。由于DNA由两条反平行线组成，并且只能以一个方向复制这两条链，因此以连续的方式合成其中一条链，领导者，而另一个则滞后，被合成为片段，随后将其组合在一起以形成最终产品。每次在滞后链上启动DNA复制时，都会发生启动事件，但是实际上，关于决定这些底漆的位置的原因几乎一无所知。我们开发了一种新的方法，用于使用下一代测序来确定启动位点在基因组水平上的位置。利用这一技术突破，我们将回答有关复制过程的基本问题，这些问题以前无法回答。我们将能够在基因组中识别复制过程减慢或终止的位置，因为在这些位置启动是特定于位置的位置。由于不频繁或过于频繁的启动事件，在复制过程中存在“问题”，我们还将能够识别区域。最后，我们将研究染色质是否会影响底漆的位置。该项目将回答有关所有细胞寿命基础的复制过程的一些非常基本的问题，并将提高我们对这一基本过程中可能出问题的知识，从而使我们深入了解癌症和衰老的基因不稳定。