Thesis
Development and optimisation of an imaging platform for interrogating RNA biology
- Creator
- Rights statement
- Awarding institution
- University of Strathclyde
- Date of award
- 2025
- Thesis identifier
- T17444
- Person Identifier (Local)
- 201771856
- Qualification Level
- Qualification Name
- Department, School or Faculty
- Abstract
- RNA plays a central role as the molecular blueprint of proteins and is a sensitive regulator of gene expression. The dysregulation of RNA processes is implicated in development of numerous diseases ranging from viral infections to cancer. By visualising these mechanisms, it is possible to identify aberrant behaviours and isolate key targets for therapeutic investigation. RNA aptamers provide a means of interrogating RNA biology with high specificity and affinity. They are versatile and open to chemical modification for optimisation of function. RNA aptamers can also be tied into the nascent procedures of molecular biology, specifically transcription and translation, to provide a live read out of expression. Finally, they are biorthogonal, allowing them to adapt to in cellulo investigations as molecular reporters. Secondary structures of RNA play functional roles in the regulation of gene expression and therefore provide a diverse model for investigating small molecule inhibition. The nascent hairpin structures found in RNA thermometers are sensitive to temperature changes that require further investigation and visualisation and their proximity to the translational start site make them ideal for twinned molecular tracking by an RNA aptamer and fluorescent protein. This thesis addresses the dynamic nature of RNA and proposes a model for the interrogation of RNA biology by applying fluorescence as a function of transcription. A comprehensive strategy for investigating the efficacy of small molecule inhibition of transcription, and optimisation of the molecular reporter used, is described. Modification of the N1 imidazolone ring (Figure 1) reports well characterized improvement over the literature standard of fluorescent dyes when bound to RNA aptamers. The development of a reporter model capable of sensitive detection of RNA structural shift is reported and provides a robust methodology of tracking temperature response in nascent RNA elements. Overall, this thesis provides valuable insight into aspects of RNA biology in need of further investigation and outlines potential utilization of the optimised platform for future research.
- Advisor / supervisor
- Burley, Glenn A.
- Hoskisson, Paul A.
- Resource Type
- DOI
- Embargo Note
Relazioni
Articoli
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File | 2025-10-28 | Embargo |