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{"title":"Synthesis of RNA Crosslinking Oligonucleotides Modified with 2-Amino-7-Deaza-7-Propynyl-6-Vinylpurine.","authors":"Ken Yamada, Yusuke Abe, Fumi Nagatsugi","doi":"10.1002/cpnc.79","DOIUrl":null,"url":null,"abstract":"<p><p>This article describes procedures to synthesize 2'-OMe-RNA modified with cross-linkable 2-amino-7-deaza-7-propynyl-6-vinylpurine (ADpVP) and preparation of the RNA-crosslinking experiment in vitro. All synthesis steps yield the desired compound in moderate or high yield without expensive chemical reagents or specific devices. The crosslink-active form of modified RNA can also be purified by commonly used reversed-phase HPLC, can be stored at -80°C after lyophilization for a few days, and is ready to use for crosslinking experiments. This crosslink-active RNA can efficiently form covalent bonds with complementary RNA in a sequence-specific manner. © 2019 by John Wiley & Sons, Inc.</p>","PeriodicalId":10966,"journal":{"name":"Current Protocols in Nucleic Acid Chemistry","volume":null,"pages":null},"PeriodicalIF":0.0000,"publicationDate":"2019-06-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1002/cpnc.79","citationCount":"0","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Current Protocols in Nucleic Acid Chemistry","FirstCategoryId":"1085","ListUrlMain":"https://doi.org/10.1002/cpnc.79","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"2019/3/12 0:00:00","PubModel":"Epub","JCR":"Q4","JCRName":"Chemistry","Score":null,"Total":0}
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Abstract
This article describes procedures to synthesize 2'-OMe-RNA modified with cross-linkable 2-amino-7-deaza-7-propynyl-6-vinylpurine (ADpVP) and preparation of the RNA-crosslinking experiment in vitro. All synthesis steps yield the desired compound in moderate or high yield without expensive chemical reagents or specific devices. The crosslink-active form of modified RNA can also be purified by commonly used reversed-phase HPLC, can be stored at -80°C after lyophilization for a few days, and is ready to use for crosslinking experiments. This crosslink-active RNA can efficiently form covalent bonds with complementary RNA in a sequence-specific manner. © 2019 by John Wiley & Sons, Inc.
2-氨基-7- deaza -7-丙基-6-乙烯嘌呤修饰RNA交联寡核苷酸的合成。
本文介绍了用可交联的2-氨基-7-二氮杂-7-丙基-6-乙烯基嘌呤(ADpVP)修饰2'-OMe-RNA的合成方法及体外交联实验的制备。所有的合成步骤都以中等或较高的产率产生所需的化合物,而不需要昂贵的化学试剂或特定的装置。修饰后的交联活性形式的RNA也可以用常用的反相高效液相色谱纯化,冻干后可在-80℃下保存数天,可用于交联实验。这种交联活性RNA可以有效地与互补RNA以序列特异性的方式形成共价键。©2019 by John Wiley & Sons, Inc。
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