Department of Cellular, Computational and Integrative Biology (CIBIO), University of Trento, Trento, Italy.
Laboratory of RNA Regulatory Networks, Department of Cellular, Computational and Integrative Biology (CIBIO), University of Trento, Trento, Italy.
Methods Mol Biol. 2022;2404:3-41. doi: 10.1007/978-1-0716-1851-6_1.
Untranslated regions of mRNA (UTRs) are involved in defining the fate of the transcript through processes such as mRNA localization, degradation, translation initiation regulation, and several others: the action of trans-factors such as RNA-binding proteins and non-coding RNAs, combined with the presence of defined sequence and structural cis-elements, ultimately determines protein synthesis levels. Identifying functional regions in UTRs and uncovering post-transcriptional regulators acting upon these is thus of paramount importance to understand this regulatory layer: these tasks can now be approached computationally to reduce the testable hypothesis space and drive the experimental validation in a more effective way.This chapter will focus on presenting databases and tools allowing to study the various aspects of post-transcriptional regulation, including the profiling of actively translated mRNAs, regulatory network analysis (e.g., RBP and ncRNA binding sites), trans-factor binding sites prediction, motif search (sequence and secondary structure), and other aspects of this regulatory layer: two potential analysis pipelines are also presented as practical examples of how these tools could be integrated and effectively employed.
mRNA 的非翻译区(UTRs)通过诸如 mRNA 定位、降解、翻译起始调节等过程参与定义转录本的命运:转录因子(如 RNA 结合蛋白和非编码 RNA)的作用,结合存在的特定序列和结构顺式元件,最终决定蛋白质合成水平。因此,鉴定 UTR 中的功能区域并揭示作用于这些区域的转录后调控因子对于理解这个调控层至关重要:现在可以通过计算方法来完成这些任务,以减少可测试的假设空间并以更有效的方式推动实验验证。这一章将重点介绍允许研究转录后调控的各个方面的数据库和工具,包括活跃翻译的 mRNAs 的分析、调控网络分析(例如,RBP 和 ncRNA 结合位点)、转录因子结合位点预测、基序搜索(序列和二级结构)以及这个调控层的其他方面:还提出了两个潜在的分析流程作为如何整合和有效利用这些工具的实际示例。