Diabetes
Understanding transcriptomic complexity of human pancreas development at the cellular level using single-cell long-read RNA sequencing.
To conduct this project, we will be combining single cell sequencing with long read RNA sequencing technologies. We isolate cells from human fetal pancreatic tissue, ensuring that we maintain the integrity of the RNA. Using the 10X Genomics Chromium platform, barcoded cDNA from each individual cell is generated. This gives each transcript a unique cell label, making it easier to analyse. We then use Oxford Nanopore long-read sequencing to read the full-length transcript.
Following the collection of the data, we will use computational tools to explore gene expression and transcript diversity across each pancreatic cell type and explore how these change during development.
This project will generate the first ever single cell long-read RNA sequencing data set in the human fetal pancreas.
The data from this project will serve as pilot data for securing a larger fellowship, aimed at profiling the human fetal pancreatic transcriptome at the single cell level across a full time-course. This study would generate a comprehensive map of isoform diversity in each cell type across pancreatic development. Additionally, data from this study will pave the way for me to conduct a complementary study focused on full-length spatial transcriptomics in the developing pancreas, leveraging our single cell long read sequencing data to provide gene expression profiles which can then be contextualized with spatial transcriptomics.
This project and subsequent studies will significantly advance our understanding of human pancreatic development.
NIHR Exeter BRC.
Nick Owens, Rachel Jennings (University of Manchester).
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