Long-read Nanopore sequencing to identify isoform-specific patterns of mRNA methylation

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It has for decades been known that the building blocks, or bases, within DNA and RNA molecules can be chemically modified. In DNA, the occurrence and biological function of such modifications has been a fundamentally important area of biosciences research with far-reaching impact for decades. In stark contrast, the progress of research into RNA modifications has been very disappointing; this is especially unfortunate as these modifications are actually known to be more prevalent and chemically complex than those found in DNA. Further, many of the enzymes that catalyse the formation of RNA modifications have been linked with human disease, suggesting important biological roles. A primary reason for slow progress in the field has been due to the fact that detailed studies identifying the precise positions of modifications in RNA molecules have proven technically very difficult. However in recent years, major advances in DNA/RNA sequencing techniques have been made that has allowed RNA modifications to be identified in the detail required to elucidate biological function. The first such studies were described in 2012 and indeed we are now beginning to realise more fully the broad scope offered by such investigations in biosciences research. The research field is however still in its early stages and a massive concerted effort is required between laboratories in order to yield necessary molecular characterizations of these RNA modifications. The studies proposed here aim to make major contributions to the field in this regard.