Investigating epitranscriptomic modification of the DMPK expansion RNA in DM1
Publication Date
July 27, 2026
Creator
Rayer, Imogen
Abstract
Myotonic dystrophy type 1 (DM1) is a rare, progressive, multisystemic disorder caused by a (CTG)n expansion in the DMPK gene. The mutant mRNA forms hairpin structures that sequester muscleblind-like proteins, leading to the formation of ribonuclear foci and dysregulated RNA metabolism. Epitranscriptomic modifications such as 5-methylcytosine (m5C) are emerging regulators of RNA metabolism and may influence DM1 pathology. Epitranscriptomic regulation is mediated by ‘writers,’ ‘readers,’ and ‘erasers’ that add, interpret, or remove RNA modifications. This study investigates the role of m5C in DM1 by targeting the ‘reader,’ ALYREF, involved in mRNA export and m5C recognition. This study also incorporates unpublished datasets on ALYREF knockdown and NSUN2 knockout, providing a broader perspective on m5C regulation in DM1.
DM1 patient-derived fibroblasts (KB-TeloMyoD) were transfected with either ALYREF-specific siRNAs, a non-targeting siRNA control or a transfection control. Cells were analysed by fluorescence in situ hybridisation (FISH). Foci area, intensity, and number per nucleus were quantified using the Zeiss CellDiscovery7 microscope, R-script, and GraphPad Prism. ALYREF knockdown was validated with western blotting.
Preliminary FISH results (n=4) show that ALYREF knockdown resulted in statistically significant but variable effects on RNA foci area, number, and intensity. NSUN2 knockout (n=2) produced an approximate two-fold reduction under similar conditions. These findings suggest that ALYREF and NSUN2, and by extension m5C, may influence RNA foci formation and stability in DM1. Future work investigating TET2 will further define epitranscriptomic contributions to DM1 pathology and may highlight novel avenues for therapeutic intervention aimed at reducing RNA toxicity.
Item Type
ethesis
Thesis Type
MRes
Supervisors
Subjects (LC)
Associated Schools / Departments
UoN Repository URI
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Rayer, Imogen, 20358213, Second.pdf
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