应用CPR改进基因共表达网络探究反式剪接的调控因子及机制

反式剪接是一种非共线的RNA剪接，可将两个独立转录本（即亲本RNA）剪接成嵌合RNA，整合异源遗传信息，多样化转录组和蛋白组。探究反式剪接的相关调控问题，可为RNA治疗提供新楔入点。然而，其调控因子及机制仍不完善。标准基因共表达网络，虽然可呈现基因间的协作，但会混杂RNA转录和RNA剪接两种调控关系，不利于探究RNA剪接机制。本组前期发现，嵌合RNA和亲本RNA在转录水上的比值（CPR），从数据标准化的角度，可剖分RNA转录与反式剪接这两种效应；将CPR作为节点，引入基因共表达网络，可改善网络的精细结构，有利于挖掘反式剪接中的调控因子及通路。因而，本项目预以CPR改进基因共表达网络，准确捕获RNA转录和反式剪接两种调控关系；以网络拓扑特性，探究反式剪接中的关键调控因子及通路；以亲和层析、分子杂交、定量分析等试验，验证所揭示规律，为反式剪接和嵌合RNA的相关研究及应用，提供新思路和新靶点。

Trans-splicing is a non-collinear RNA splicing that fuses two independent transcripts (ie, parental RNAs) into a chimeric RNA, which in turn can integrate heterologous genetic information and diversify transcriptomes and proteomes. To explore its related regulatory issues can improve trans-splicing-mediated RNA therapy. However, its regulatory factors and mechanisms remain elusive. Although the standard gene co-expression network can high-throughput show inter-genetic collaboration, it will mix the regulations in two areas of RNA transcription and RNA splicing, which is not conducive to exploring RNA splicing mechanism. In our previous study, we found that the ratio of chimeric RNA to parental RNA on the transcription level (CPR) can divide the two parts: RNA transcription and trans-splicing in terms of data standardization. Using CPRs as nodes can improve the fine structure of the network and facilitate the mining of regulators and pathways in trans-splicing. Therefore, the present project will use CPRs to improve gene co-expression network, simultaneously capturing two kinds of regulatory relationship of RNA transcription and trans-splicing. We will then use network topological characteristics to identify key regulatory factors and pathways in trans-splicing. We will use affinity chromatography, molecular hybridization and quantitative analysis to verify the revealed rules. Finally, we seek to provide new ideas and targets in related researches and applications of trans-splicing and chimeric RNA.