{"title":"RNA aptamers as tools for the purification and analysis of in vivo assembled Ribonucleoproteins.","authors":"Daniel G Rocca, Ute Kothe","doi":"10.1261/rna.080460.125","DOIUrl":null,"url":null,"abstract":"<p><p>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 utilized 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.</p>","PeriodicalId":21401,"journal":{"name":"RNA","volume":" ","pages":""},"PeriodicalIF":4.2000,"publicationDate":"2025-06-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"RNA","FirstCategoryId":"99","ListUrlMain":"https://doi.org/10.1261/rna.080460.125","RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q2","JCRName":"BIOCHEMISTRY & MOLECULAR BIOLOGY","Score":null,"Total":0}
引用次数: 0
Abstract
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 utilized 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.
期刊介绍：
RNA is a monthly journal which provides rapid publication of significant original research in all areas of RNA structure and function in eukaryotic, prokaryotic, and viral systems. It covers a broad range of subjects in RNA research, including: structural analysis by biochemical or biophysical means; mRNA structure, function and biogenesis; alternative processing: cis-acting elements and trans-acting factors; ribosome structure and function; translational control; RNA catalysis; tRNA structure, function, biogenesis and identity; RNA editing; rRNA structure, function and biogenesis; RNA transport and localization; regulatory RNAs; large and small RNP structure, function and biogenesis; viral RNA metabolism; RNA stability and turnover; in vitro evolution; and RNA chemistry.
求助内容：
应助结果提醒方式：
京公网安备 11010802042870号
