{"title":"Visualizing RNA Cytidine Acetyltransferase Activity by Northern Blotting","authors":"Keri M. Bryson, Supuni Thalalla-Gamage, Jordan L. Meier","doi":"10.1002/cpch.89","DOIUrl":null,"url":null,"abstract":"<p>Cytidine acetyltransferases are an emerging class of nucleic-acid-modifying enzymes responsible for the establishment of <i>N</i><sup>4</sup>-acetylcytidine (ac4C) in RNA. In contrast to histone acetyltransferases, whose activity is commonly studied by western blotting, relatively few methods exist for quickly assessing the activity of cytidine acetyltransferases from a biological sample of interest or the distribution of ac4C across different RNA species. In this protocol, we describe a method for analysis of cellular cytidine acetyltransferase activity using dot- and immuno-northern-blotting-based detection. © 2020 Wiley Periodicals LLC.</p><p><b>Basic Protocol 1</b>: Detection of <i>N</i><sup>4</sup>-Acetylcytidine in RNA by dot blotting</p><p><b>Basic Protocol 2</b>: Visualizing <i>N</i><sup>4</sup>-Acetylcytidine Distribution in RNA by northern blotting</p>","PeriodicalId":38051,"journal":{"name":"Current protocols in chemical biology","volume":"12 4","pages":""},"PeriodicalIF":0.0000,"publicationDate":"2020-12-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1002/cpch.89","citationCount":"1","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Current protocols in chemical biology","FirstCategoryId":"1085","ListUrlMain":"https://onlinelibrary.wiley.com/doi/10.1002/cpch.89","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q3","JCRName":"Biochemistry, Genetics and Molecular Biology","Score":null,"Total":0}
引用次数: 1
Abstract
Cytidine acetyltransferases are an emerging class of nucleic-acid-modifying enzymes responsible for the establishment of N4-acetylcytidine (ac4C) in RNA. In contrast to histone acetyltransferases, whose activity is commonly studied by western blotting, relatively few methods exist for quickly assessing the activity of cytidine acetyltransferases from a biological sample of interest or the distribution of ac4C across different RNA species. In this protocol, we describe a method for analysis of cellular cytidine acetyltransferase activity using dot- and immuno-northern-blotting-based detection. © 2020 Wiley Periodicals LLC.
Basic Protocol 1: Detection of N4-Acetylcytidine in RNA by dot blotting
Basic Protocol 2: Visualizing N4-Acetylcytidine Distribution in RNA by northern blotting
用Northern Blotting观察RNA胞苷乙酰转移酶活性
胞苷乙酰转移酶是一类新兴的核酸修饰酶，负责在RNA中建立n4 -乙酰胞苷(ac4C)。与通常通过western blotting研究活性的组蛋白乙酰转移酶相比，相对较少的方法可以从感兴趣的生物样品中快速评估胞苷乙酰转移酶的活性或ac4C在不同RNA物种中的分布。在本协议中，我们描述了一种方法，分析细胞胞苷乙酰转移酶活性使用点和免疫北方印迹为基础的检测。©2020 Wiley期刊公司。基本方案1:用点印迹法检测RNA中n4 -乙酰胞苷基本方案2:用northern印迹法可视化RNA中n4 -乙酰胞苷的分布
本文章由计算机程序翻译，如有差异，请以英文原文为准。
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