The biology of DNA triplexes in the model genetic organisms C. elegans and Drosophila

模式遗传生物线虫和果蝇中 DNA 三链体的生物学

• 批准号: BB/D005981/1
• 负责人: Keith R Fox
• 金额: $ 71.82万
• 依托单位: University of Southampton
• 依托单位国家: 英国
• 项目类别: Research Grant
• 财政年份: 2006
• 资助国家: 英国
• 起止时间: 2006 至 无数据
• 项目状态: 已结题
• 来源: https://gtr.ukri.org/projects?ref=BB%2FD005981%2F1
• 关键词: biology; DNA; triplexes; model; genetic

The genome sequencing projects have provided the raw material for the scientific community to investigate the genetic basis of the normal physiological function of cells and their dysfunction in disease. To pursue this, new experimental tools are also required, especially to modulate the expression of individual genes in any cell and to define the consequence of these interventions. Ultimately, there is also the hope that such approaches may be used in clinical situations e.g. in the treatment of genetic diseases and cancers. Several of these approaches employ synthetic oligonucleotides. These are molecules either resemble the native chromosomal DNA, i.e. the genetic code, or the RNA which carries the genetic signal inside the cell. A particularly powerful method to regulate gene expression uses molecules which are designed to bind to DNA. These are modified oligonucleotides, called triplex-forming oligonucleotides (TFOs), which bind to the double-stranded DNA to form a triplex structure which prevents the gene from being expressed. Furthermore, in some instances this triplex structure can induce permanent genetic mutations. There is considerable interest in developing these molecules as a mechanism for regulating gene expression. Despite this interest however, relatively little is known about the effects of TFOs on gene expression in intact organisms. This project has put together a team of chemists and biologists keen to address this problem. They will systematically test and characterise the effects of TFOs in the model genetic animals the nematode worm, C. elegans and the fruit fly, Drosophila. Using these animals as models has tremendous advantages, especially because there is already a wealth of information on genetic mutants on which to base these studies. Essentially the project will endeavour to mimic the phenotypes observed in genetic mutants by treating wild-type animals with TFOs. Importantly, the effect of the TFOs will also be carefully characterised so that information can be obtained on how effectively they regulate gene expression. The project will also provide information on other features of the effects of TFOs in a physiological environment which are currently unknown, such as how specific and persistent the effects are, particularly in the context of the developing organism during cell division and growth. The net result of this will be an important advance in the understanding of the mechanism of action of gene regulation by modified oligonucleotides. This will be benefit both those in the scientific community who are looking for new tools to interpret the functional meaning of the animal genome sequences, and also those who wish to exploit this technology for the treatment of human disease.

基因组测序计划为科学界研究细胞正常生理功能及其在疾病中功能障碍的遗传基础提供了原始材料。为了实现这一目标，还需要新的实验工具，特别是调节任何细胞中单个基因的表达，并确定这些干预措施的后果。最终，还希望这些方法可以用于临床情况，例如治疗遗传疾病和癌症。这些方法中的几种使用合成的寡核苷酸。这些分子要么类似于天然染色体DNA，即遗传密码，要么类似于携带细胞内遗传信号的RNA。调节基因表达的一种特别有效的方法是使用被设计成与DNA结合的分子。这些是修饰的寡核苷酸，称为三链体形成寡核苷酸（TFO），其结合到双链DNA以形成阻止基因表达的三链体结构。此外，在某些情况下，这种三链体结构可以诱导永久性基因突变。人们对开发这些分子作为调节基因表达的机制有相当大的兴趣。然而，尽管有这种兴趣，对TFO对完整生物体中基因表达的影响知之甚少。这个项目汇集了一个化学家和生物学家的团队，他们热衷于解决这个问题。他们将系统地测试和验证TFO在模式遗传动物线虫、C. elegans线虫and the fruit水果fly飞，Drosophila果蝇.使用这些动物作为模型具有巨大的优势，特别是因为已经有大量关于遗传突变的信息作为这些研究的基础。基本上，该项目将努力通过用TFO治疗野生型动物来模拟在遗传突变体中观察到的表型。重要的是，TFO的作用也将被仔细表征，以便获得关于它们如何有效地调节基因表达的信息。该项目还将提供有关目前未知的TFO在生理环境中影响的其他特征的信息，例如影响的特异性和持久性，特别是在细胞分裂和生长期间发育中的生物体中。这将是一个重要的进展，在理解的作用机制的基因调控修饰的寡核苷酸的净结果。这将有利于科学界那些正在寻找新工具来解释动物基因组序列的功能意义的人，以及那些希望利用这项技术治疗人类疾病的人。

项目成果

DNA triplex formation with 5-dimethylaminopropargyl deoxyuridine.

• DOI: 10.1093/nar/gkn1060
• 发表时间: 2009-03
• 期刊: Nucleic acids research
• 影响因子: 14.9
• 作者: Rusling DA;Peng G;Srinivasan N;Fox KR;Brown T
• 通讯作者: Brown T

Towards the Targeted Modulation of Gene Expression by Modified Triplex-Forming Oligonucleotides

通过修饰的三链体形成寡核苷酸靶向调节基因表达

• DOI: 10.2174/187231308783334135
• 发表时间: 2008
• 期刊: Current Chemical Biology
• 作者: Fox K