RNA interaction studies : a pathway to the development of a surface-based technology

• Main Author: Phillips, Jack O.
• Other Authors: Callaghan, Anastasia Jane
• Published: University of Portsmouth 2016
• Subjects: 572.8
• Online Access: https://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.714201

Ribonucleic acid (RNA) has long been thought of as the bearer and the machinery of translating the genetic code. It is now known that RNA is capable of diverse functions both in a cellular and in a synthetic context which have highlighted RNA as an important biological molecule to be studied and as a potential toolkit for the researcher. Research into RNA has uncovered a wealth of subgroups of RNA with different functions including the bacterial small non-coding RNAs (sRNAs). These sRNAs and other RNA subgroups are now known to be key cellular regulators of important activities such as virulence and stress response and thus emphasise their importance in antibiotic research. Many RNAs regulate their function and the function of other biomolecules by specific interactions with other RNAs, proteins, small molecules and even ions. However, identifying specific meaningful RNA interactions and their consequences is troublesome and methods to study these interactions are slow and laborious. This work has focused on developing protocols for applying existing and novel techniques to studying important RNA interactions which has led to the invention of a new technology for creating functional multi RNA arrays. Initial work employed structural and binding analyses to investigate the details of three importants RNA interactions with divalent ions or the chaperone protein Hfq, which provided clarity on previous data. Subsequently, efforts were made to develop methods to studying RNA interactions in detail but with medium throughput with surface plasmon resonance imaging, not previously used for studying RNA interactions. Finally, protocols were developed to create stable and functional, multi RNA arrays that were subsequently validated for use in screening RNA interactions in high-throughput with the demonstration of specific RNA-RNA, RNA-protein and RNA-small molecule interactions. This work sets the precedent for the utilisation of higher throughput methods in studying RNA interactions towards the greater understanding and exploitation of RNA functions and capabilities.