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{"title":"Copper-Catalyzed Huisgen 1,3-Dipolar Cycloaddition Tailored for Phosphorothioate Oligonucleotides","authors":"Malgorzata Honcharenko, Dmytro Honcharenko, Roger Stromberg","doi":"10.1002/cpnc.102","DOIUrl":null,"url":null,"abstract":"<p>An efficient method for attachment of a variety of reporter groups to oligonucleotides (ONs) is copper (I) [Cu(I)]-catalyzed Huisgen azide-alkyne 1,3-dipolar cycloaddition (“click reaction”). However, in the case of ONs with phosphorothioate modifications as internucleosidic linkages (PS-ONs), this conjugation method has to be adjusted to be compatible with the sulfur-containing groups. The method described here is adapted for PS-ONs, utilizes solid-supported ONs, and implements the Cu(I) bromide dimethyl sulfide complex (CuBr × Me<sub>2</sub>S) as a mediator for the click reaction. The solid-supported ONs can be readily transformed into “clickable ONs” by on-line addition of an alkyne-containing linker that subsequently can react with an azido-containing moiety (e.g., a peptide) in the presence of CuBr × Me<sub>2</sub>S. © 2019 by John Wiley & Sons, Inc.</p><p><b>Basic Protocol 1</b>: Conjugation on solid support</p><p><b>Support Protocol</b>: Removal of 4,4′-dimethoxytrityl group from amino linker</p><p><b>Basic Protocol 2</b>: Removal of protecting groups and cleavage from solid support</p><p><b>Basic Protocol 3</b>: HPLC purification</p>","PeriodicalId":10966,"journal":{"name":"Current Protocols in Nucleic Acid Chemistry","volume":null,"pages":null},"PeriodicalIF":0.0000,"publicationDate":"2019-12-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1002/cpnc.102","citationCount":"1","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Current Protocols in Nucleic Acid Chemistry","FirstCategoryId":"1085","ListUrlMain":"https://onlinelibrary.wiley.com/doi/10.1002/cpnc.102","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q4","JCRName":"Chemistry","Score":null,"Total":0}
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Abstract
An efficient method for attachment of a variety of reporter groups to oligonucleotides (ONs) is copper (I) [Cu(I)]-catalyzed Huisgen azide-alkyne 1,3-dipolar cycloaddition (“click reaction”). However, in the case of ONs with phosphorothioate modifications as internucleosidic linkages (PS-ONs), this conjugation method has to be adjusted to be compatible with the sulfur-containing groups. The method described here is adapted for PS-ONs, utilizes solid-supported ONs, and implements the Cu(I) bromide dimethyl sulfide complex (CuBr × Me2 S) as a mediator for the click reaction. The solid-supported ONs can be readily transformed into “clickable ONs” by on-line addition of an alkyne-containing linker that subsequently can react with an azido-containing moiety (e.g., a peptide) in the presence of CuBr × Me2 S. © 2019 by John Wiley & Sons, Inc.
Basic Protocol 1 : Conjugation on solid support
Support Protocol : Removal of 4,4′-dimethoxytrityl group from amino linker
Basic Protocol 2 : Removal of protecting groups and cleavage from solid support
Basic Protocol 3 : HPLC purification
专为硫代寡核苷酸设计的铜催化Huisgen 1,3-偶极环加成
铜(I) [Cu(I)]催化的Huisgen叠氮- 1,3-炔偶极环加成反应(“click反应”)是一种将多种报告基团连接到寡核苷酸(ONs)上的有效方法。然而，对于具有硫代修饰的核苷间键(PS-ONs)的离子，必须调整这种缀合方法以与含硫基团相容。本文描述的方法适用于PS-ONs，利用固体负载的ONs，并实现了Cu(I)溴化二甲基硫化配合物(CuBr × Me2S)作为点击反应的介质。通过在线添加含炔的连接剂，固体支撑的离子可以很容易地转化为“可点击的离子”，该连接剂随后可以在CuBr × Me2S存在下与含叠氮基团的部分(例如肽)反应。©2019 by John Wiley &基本方案1:固体支架上的偶联;支持方案:从氨基连接物上去除4,4 ' -二甲氧基三硝基;基本方案2:从固体支架上去除保护基团和裂解;基本方案3:高效液相色谱纯化
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