Howard Hughes Medical Institute, Department of Biology, Brandeis University, Waltham, MA 02451, USA.
Department of Anatomy and Structural Biology, Albert Einstein College of Medicine, Bronx, NY 10461, USA.
Mol Cell. 2022 Jan 20;82(2):389-403. doi: 10.1016/j.molcel.2021.10.010. Epub 2021 Nov 4.
RNA binding proteins (RBPs) regulate nearly all post-transcriptional processes within cells. To fully understand RBP function, it is essential to identify their in vivo targets. Standard techniques for profiling RBP targets, such as crosslinking immunoprecipitation (CLIP) and its variants, are limited or suboptimal in some situations, e.g. when compatible antibodies are not available and when dealing with small cell populations such as neuronal subtypes and primary stem cells. This review summarizes and compares several genetic approaches recently designed to identify RBP targets in such circumstances. TRIBE (targets of RNA binding proteins identified by editing), RNA tagging, and STAMP (surveying targets by APOBEC-mediated profiling) are new genetic tools useful for the study of post-transcriptional regulation and RBP identification. We describe the underlying RNA base editing technology, recent applications, and therapeutic implications.
RNA 结合蛋白（RBPs）调节细胞内几乎所有的转录后过程。为了全面了解 RBP 的功能，识别它们的体内靶标至关重要。用于分析 RBP 靶标的标准技术，如交联免疫沉淀（CLIP）及其变体，在某些情况下受到限制或不理想，例如当没有相容的抗体时，以及当处理小细胞群体，如神经元亚型和原代干细胞时。本综述总结并比较了几种最近设计的遗传方法，用于在这种情况下鉴定 RBP 靶标。TRIBE（通过编辑鉴定的 RNA 结合蛋白的靶标）、RNA 标记和 STAMP（通过 APOBEC 介导的分析进行目标调查）是新的遗传工具，可用于研究转录后调控和 RBP 鉴定。我们描述了基本的 RNA 碱基编辑技术、最近的应用以及治疗意义。
