Rocca Daniel G, Kothe Ute
Department of Chemistry, University of Manitoba, Winnipeg, Manitoba, R3T 2N2 Canada.
Department of Chemistry, University of Manitoba, Winnipeg, Manitoba, R3T 2N2 Canada
RNA. 2025 Aug 18;31(9):1235-1247. doi: 10.1261/rna.080460.125.
A large number of ribonucleoprotein (RNP) complexes are being discovered mediating numerous cellular functions. To investigate the composition, structure, and functional mechanism of RNP complexes, it is advantageous to isolate an RNP that was assembled in vivo. This review provides a systematic overview of a versatile and highly effective method to accomplish this task, namely, the purification of RNPs from cells using genetically encoded RNA aptamers. Inserting an RNA aptamer into the RNA of an RNP enables binding of the tagged RNP with high affinity and specificity to a ligand as an effective affinity chromatography purification strategy. Therefore, the purification of RNPs using aptamers has been used successfully to identify heterogenous populations of RNPs forming around a single RNA as well as to characterize intermediates in the formation of complex RNPs such as the ribosome. Here, we discuss in detail the selection of an appropriate RNA aptamer based on the properties of both the aptamer and its ligand, and we describe critical considerations in designing RNP purifications.
大量核糖核蛋白（RNP）复合物被发现介导着众多细胞功能。为了研究RNP复合物的组成、结构和功能机制，分离在体内组装的RNP是有利的。本综述系统概述了一种通用且高效的方法来完成这项任务，即使用基因编码的RNA适体从细胞中纯化RNP。将RNA适体插入RNP的RNA中，能够使标记的RNP与配体以高亲和力和特异性结合，作为一种有效的亲和色谱纯化策略。因此，使用适体纯化RNP已成功用于鉴定围绕单个RNA形成的异质性RNP群体，以及表征复杂RNP（如核糖体）形成过程中的中间体。在这里，我们根据适体及其配体的特性详细讨论合适RNA适体的选择，并描述设计RNP纯化时的关键注意事项。
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