Identifying transcriptome diversity and RNA modifications across human pancreatic development

Genetics and Genomics

Summary

Mapping gene expression regulation in the human fetal pancreas using RNA sequencing to uncover novel features and modifications.

How are we doing it?

This project will be conducted in a series of key stages.

In the lab, RNA is extracted from human fetal pancreas tissue. Because of the nature of this sequencing technique, we ensure that the RNA is of high quality. Following extraction, we check the quantity and quality of the RNA. We then perform library preparation of these samples, in brief, this is a stage to ensure that the RNA is ready to be sequenced. Finally, we sequence the native RNA using Oxford Nanopore.

Once the RNA is sequenced, we then use standard Oxford nanopore analysis pipelines alongside our own bespoke analysis to explore RNA modifications and transcriptome diversity across pancreatic development. We will, in particular, observe which genes have dynamic developmental RNA modifications and transcript usage.

What happens next?

This project is the first application of native RNA sequencing in the developing human pancreas. The data we will obtain will provide an unprecedented view of the human fetal pancreatic transcriptome. With this data, we will generate a map of transcriptomic diversity and RNA modifications across pancreatic development. This resource will enable the reinterpretation of seemingly non-coding variants associated with pancreatic disease. By directly measuring RNA modifications occurring across pancreas development, we open new avenues for understanding how genetic variants can impact these modifications and therefore impact RNA stability, splicing and translational efficiency.

With the establishment of this developmental map, we then plan to collaborate with geneticists to interpret disease associated variants.

The data and insights from this project will support future grant applications in which we aim to scale up this project to include a broader range of developmental timepoints to gain a clearer picture of transcriptomic changes in the pancreas. In parallel, we would plan to begin exploring the functional role of these RNA modifications and novel transcripts in the pancreas.

Funding

NIHR Exeter BRC.

People Involved

Dr Nick Owens.