RNA Biology and Molecular Physiology, Faculty of Biology, Bielefeld University, 33615 Bielefeld, Germany.
RNA Biology and Molecular Physiology, Faculty of Biology, Bielefeld University, 33615 Bielefeld, Germany.
Trends Plant Sci. 2018 Apr;23(4):352-365. doi: 10.1016/j.tplants.2018.01.004. Epub 2018 Feb 9.
Research into the regulation of gene expression underwent a shift from focusing on DNA-binding proteins as key transcriptional regulators to RNA-binding proteins (RBPs) that come into play once transcription has been initiated. RBPs orchestrate all RNA-processing steps in the cell. To obtain a global view of in vivo targets, the RNA complement associated with particular RBPs is determined via immunoprecipitation of the RBP and subsequent identification of bound RNAs via RNA-seq. Here, we describe technical advances in identifying RBP in vivo targets and their binding motifs. We provide an up-to-date view of targets of nucleocytoplasmic RBPs collected in arabidopsis. We also discuss current experimental limitations and provide an outlook on how the approaches may advance our understanding of post-transcriptional networks.
对基因表达调控的研究经历了一个从关注 DNA 结合蛋白作为关键转录调控因子到 RNA 结合蛋白(RBPs)的转变,后者在转录起始后发挥作用。RBPs 协调细胞内的所有 RNA 加工步骤。为了获得体内靶标的全局视图,通过 RBP 的免疫沉淀和随后通过 RNA-seq 鉴定结合的 RNA 来确定与特定 RBPs 相关的 RNA 成分。在这里,我们描述了鉴定体内 RBP 靶标及其结合基序的技术进展。我们提供了在拟南芥中收集的核质 RBPs 靶标的最新视图。我们还讨论了当前的实验限制,并展望了这些方法如何促进我们对转录后网络的理解。