The role of microRNAs and their targets in CD8+ T cell responses

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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.