{"title":"用Northern Blotting观察RNA胞苷乙酰转移酶活性","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":"{\"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}","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}
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