The role of microRNAs and their targets in CD8+ T cell responses
Lead Research Organisation:
University of Edinburgh
Department Name: Sch of Biological Sciences
Abstract
MicroRNAs (miRNAs) play an important role in immune cell function and affect many aspects of T cell immunity. In order to understand how miRNAs regulate T cell development, differentiation and activation, it is essential to know their targets. Despite improvements in bioinformatic prediction of miRNA targets, it remains challenging to identify those that are biologically relevant and to differentiate between direct and indirect targets. Recently a novel method called CLASH (cross-linking, ligation and sequencing of hybrids) has been developed to study RNA-RNA interactions. MiRNA-mRNA pairs from the RNA-induced silencing complex (RISC) can be purified from cells expressing N-terminally tagged Argonaute protein. Interacting miRNA-mRNA pairs are trimmed, ligated, amplified and sequenced. MiRNAs and their targets can then be identified as "chimeric" reads that are non-contiguous in the transcriptome. We will be using CLASH to study miRNA-mRNA interactions in CD8+ T cells. Gaining information about miRNA targets will improve our knowledge of the mechanism of action of miRNAs as well as the signalling pathways surrounding T cell activation.
Organisations
People |
ORCID iD |
| Rose Zamoyska (Primary Supervisor) |
Studentship Projects
| Project Reference | Relationship | Related To | Start | End | Student Name |
|---|---|---|---|---|---|
| BB/M010996/1 | 01/10/2015 | 31/03/2024 | |||
| 1598868 | Studentship | BB/M010996/1 | 01/10/2015 | 30/09/2019 |
| Description | This award funded the work that was reported in my PhD thesis titled 'The identification of RISC-associated microRNAs and their targets during CD8+ T cell activation'. The key findings from this work included furthering our understanding of the formation of the RNA-induced silencing complex (RISC) and microRNA function in regulating gene expression in CD8+ T cells, complementing existing work in the field. The project aims also included identification of microRNA targets by a novel biochemical method called CLASH (for Cross-linking, ligation and sequencing of hybrids). This proved challenging but after optimisation of the protocol, data were generated which allowed identification of putative microRNA targets in T cells. |
| Exploitation Route | Two large dataset were generated from this work, from which potentially important microRNAs and their targets in CD8+ T cells can be identified. While I focused on the validation of one such microRNA, further potentially interesting candidates could be explored by others. Some of the work regarding identification of microRNA targets has been taken forward by other lab members. Since microRNAs are known to play an important role in immune cell function, and dysregulation of microRNAs can be seen in many human diseases including autoimmunity and cancer, potential novel therapeutic strategies could be developed based on promoting or inhibiting specific microRNAs in T cells. For such applications it is crucial to know the precise roles and targets of microRNAs in T cells. |
| Sectors | Pharmaceuticals and Medical Biotechnology |