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

基因组测序项目为科学界研究细胞正常生理功能及其在疾病中的功能障碍的遗传基础提供了原料。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的后果。最终，也有希望将这些方法用于临床情况，例如遗传性疾病和癌症的治疗。这些方法中有几种采用合成寡核苷酸。这些分子要么类似于天然染色体DNA，即遗传密码，要么类似于细胞内携带遗传信号的RNA。一种特别有效的调节基因表达的方法是使用与DNA结合的分子。这些是修饰过的寡核苷酸，称为三联体形成寡核苷酸（TFOs），它们与双链DNA结合形成三联体结构，阻止基因表达。此外，在某些情况下，这种三重结构可以诱导永久的基因突变。人们对开发这些分子作为调节基因表达的机制非常感兴趣。然而，尽管有这种兴趣，对TFOs对完整生物体中基因表达的影响知之甚少。这个项目将化学家和生物学家组成了一个团队，致力于解决这个问题。他们将系统地测试和描述全氟辛烷磺酸在模型遗传动物——线虫、秀丽隐杆线虫和果蝇中的影响。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基因突变的信息可以作为这些研究的基础。从本质上讲，该项目将努力通过用TFOs治疗野生型动物来模仿在基因突变中观察到的表型。重要的是，tfo的作用也将被仔细地描述，以便获得它们如何有效地调节基因表达的信息。该项目还将提供关于目前未知的生理环境中全氟辛烷磺酸影响的其他特征的资料，例如影响的特异性和持久性，特别是在细胞分裂和生长过程中发育中的生物体。这项研究的最终结果将是对修饰寡核苷酸基因调控作用机制理解的重要进展。这将有利于科学界寻找新工具来解释动物基因组序列的功能意义的人，也有利于那些希望利用这项技术治疗人类疾病的人。

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.