Development of analytical approaches in the analysis of RNA

RNA 分析方法的发展

• 批准号: EP/D033713/1
• 负责人: Mark Dickman
• 金额: $ 15.9万
• 依托单位: University of Sheffield
• 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=EP%2FD033713%2F1
• 关键词: Development; analytical; approaches; analysis; RNA

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

生物RNA的分离和表征方法的发展与其大分子对应物DNA和蛋白质的分析方法的发展有着某种程度的曲折历史。此外，RNA的提取、分离和分析通常比DNA所需的更困难。在处理RNA分离问题时，稳定性是直接关注的问题。RNA容易降解，使得提取的初始阶段和纯化材料的下游储存比DNA更具挑战性。从细胞中提取的总RNA含有两种最丰富的种类，对应于两种主要RNA（核糖体RNA）。其他主要RNA分子包括物种特异性转移RNA和较小的核糖体RNA种类。最近，许多研究的焦点已经被执行到一组小RNA物种（称为小核RNA）所扮演的角色。这些小RNA的长度约为22个核苷酸，通常用于调节基因表达。这种RNA的分离和分析仍然存在问题。鉴定这种RNA种类的重要性日益增加，需要开发技术来克服与当前方法相关的许多注意事项。高质量小RNA的富集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。在此背景下，提出开发RNA分析的分析方法。RNA分析新技术的发展及其在miRNA研究中的应用是多学科实验计划的理想论坛，包括生物系统分析中的分析技术方面。这项研究将在谢菲尔德大学化学与过程工程系的系统生物学小组内进行

项目成果

Purification and characterisation of dsRNA using ion pair reverse phase chromatography and mass spectrometry.

使用离子对反相色谱和质谱法对DSRNA的纯化和表征。

• DOI: 10.1016/j.chroma.2016.12.062
• 发表时间: 2017-02-10
• 期刊: Journal of chromatography. A
• 作者: Nwokeoji AO;Kung AW;Kilby PM;Portwood DE;Dickman MJ
• 通讯作者: Dickman MJ

Nucleic acid separations using superficially porous silica particles.

• DOI: 10.1016/j.chroma.2016.02.057
• 发表时间: 2016-04-01
• 期刊: Journal of chromatography. A
• 作者: Close ED;Nwokeoji AO;Milton D;Cook K;Hindocha DM;Hook EC;Wood H;Dickman MJ
• 通讯作者: Dickman MJ

Sequestration of multiple RNA recognition motif-containing proteins by C9orf72 repeat expansions.

• DOI: 10.1093/brain/awu120
• 发表时间: 2014-07
• 期刊: Brain : a journal of neurology
• 作者: Cooper-Knock J;Walsh MJ;Higginbottom A;Robin Highley J;Dickman MJ;Edbauer D;Ince PG;Wharton SB;Wilson SA;Kirby J;Hautbergue GM;Shaw PJ
• 通讯作者: Shaw PJ

Small CRISPR RNAs guide antiviral defense in prokaryotes.

• DOI: 10.1126/science.1159689
• 发表时间: 2008-08-15
• 期刊: Science (New York, N.Y.)
• 作者: Brouns SJ;Jore MM;Lundgren M;Westra ER;Slijkhuis RJ;Snijders AP;Dickman MJ;Makarova KS;Koonin EV;van der Oost J
• 通讯作者: van der Oost J

Ion Pair Reverse Phase Chromatography: A Versatile Platform for the Analysis of RNA

离子对反相色谱：RNA 分析的多功能平台

• 期刊: Chromatography Today
• 作者: Mark Dickman (Author)