Development of analytical approaches in the analysis of RNA

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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