Development of analytical approaches in the analysis of RNA

生物核糖核酸的分离和表征方法的发展，与分析其大分子对应DNA和蛋白质的类似方法一样，经历了一段有点摇摆不定的历史。此外，与DNA相比，RNA的提取、分离和分析通常更困难。在处理RNA分离问题时，稳定性是当务之急。RNA很容易降解，这使得提取的初始阶段和纯化材料的下游存储比DNA更具挑战性。从细胞中提取的总RNA包含两个最丰富的物种，对应于两个主要的RNA(核糖体RNA)。其他主要的RNA分子包括物种特异性转移RNA和较小的核糖体RNA物种。最近，许多研究的重点是一组被称为小核RNA的小RNA物种所起的作用。这些小RNA的长度约为22个核苷酸，通常起着调节基因表达的作用。对这类RNA的分离和分析仍然存在问题。在鉴定这类RNA物种方面日益重要，需要开发技术来克服与当前方法相关的许多警告。高质量小RNA的浓缩和分离以及快速分离机制和高灵敏度是本提案希望解决的主题。在这种背景下，建议开发分析方法来分析RNA。RNA分析新技术的发展及其在miRNA研究中的应用是一个多学科实验方案的理想论坛，该方案涵盖了生物系统分析中分析技术的各个方面。这项研究将在谢菲尔德大学化学和过程工程系的系统生物学小组内进行

The development of methodology in the isolation and characterisation of biological RNA has had a somewhat chequered history alongside comparable methods for the analysis of its macromolecular counterparts DNA and proteins. Moreover the extraction, isolation and analysis of RNA is routinely more difficult in comparison to that required for DNA. In approaching the problem of RNA isolation the stability is of immediate concern. RNA is susceptible to degradation, rendering the initial stages of extraction and the downstream storage of the purified material more challenging than for DNA. Total RNA extracted from cells contain two most abundant species corresponding to the two major RNAs (ribosomal RNAs). The other major RNA molecules include a species specific transfer RNA and smaller ribosomal RNA species. More recently the focus of much research has been performed into the roles played by a group of small RNA species referred to as small nuclear RNAs . These small RNAs are about 22 nucleotides in length and often function to regulate gene expression. The isolation and analysis of such RNAs remains problematic. The increasing importance in the identification of such RNA species requires the development of technology to overcome many of the caveats associated with current methods. The enrichment and isolation of high quality small RNAs in conjunction with rapid separation mechanisms and high sensitivity is the subject that this proposal wishes to address.Against this background it is proposed to develop analytical methodology in the analysis of RNA. The development of novel techniques in RNA analysis and its application in the study of miRNA is an ideal forum for a multi-disciplinary experimental programme, encompassing aspects of analytical techniques in the analysis of biological systems. The research will be performed within the Systems Biology group in the Department of Chemical and Process Engineering at the University of Sheffield