SITE-SPECIFIC LABELLING OF RNA BY THE SYNTHESIS OF NON-NATURAL BASE-PAIRS

The aim of this studentship is to explore the capacity to site-specifically label RNA by an expanded genetic repertoire. The ultimate goal of this work is to establish the first example of enzymatic incorporation of two functional groups in pre-mRNA that can be labelled with fluorophores for use in FRET-based analysis of RNA biology.
This work will involve the development of synthetic methodology to produce tagged versions of non-natural base-pairs known as Ds and Pa. These non-natural base-pairs are known to pair with one another but poorly with naturally-occurring nucleotides. The first phase of the project will involve the development of efficient synthetic routes for the preparation of modified Ds and Pa. In the second phase of the project, the conditions for efficient transcription and site-specific fluorophore labelling of mRNA containing Da and Pa will be determined. In the final phase of the project, these methods will be employed to investigate how the structural dynamics of RNA plays a role in the splicing of the oncogene Bcl-x.