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

microRNA（miRNAs）在免疫细胞功能中起重要作用，并影响T细胞免疫的许多方面。为了了解miRNAs如何调节T细胞的发育、分化和活化，了解它们的靶点至关重要。尽管miRNA靶点的生物信息学预测有所改进，但识别那些生物学相关的靶点并区分直接和间接靶点仍然具有挑战性。最近，一种称为CLASH（cross-linking，ligation and sequencing of hybrids）的新方法被开发用于研究RNA-RNA相互作用。来自RNA诱导的沉默复合物（RISC）的MiRNA-mRNA对可以从表达N-末端标记的Argonaute蛋白的细胞中纯化。修剪、连接、扩增和测序相互作用的miRNA-mRNA对。然后可以将miRNA及其靶标鉴定为在转录组中不连续的“嵌合”读段。我们将使用CLASH来研究CD 8 + T细胞中的miRNA-mRNA相互作用。获得有关miRNA靶点的信息将提高我们对miRNA作用机制以及T细胞活化周围信号通路的认识。

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.