{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,1,23]],"date-time":"2026-01-23T07:04:21Z","timestamp":1769151861866,"version":"3.49.0"},"reference-count":25,"publisher":"Wiley","issue":"1","license":[{"start":{"date-parts":[[2020,7,27]],"date-time":"2020-07-27T00:00:00Z","timestamp":1595808000000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"funder":[{"DOI":"10.13039\/501100004442","name":"Narodowym Centrum Nauki","doi-asserted-by":"publisher","award":["UMO\u20102017\/24\/T\/NZ1\/00345"],"award-info":[{"award-number":["UMO\u20102017\/24\/T\/NZ1\/00345"]}],"id":[{"id":"10.13039\/501100004442","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100004442","name":"Narodowym Centrum Nauki","doi-asserted-by":"publisher","award":["UMO\u20102016\/21\/B\/ST5\/02556"],"award-info":[{"award-number":["UMO\u20102016\/21\/B\/ST5\/02556"]}],"id":[{"id":"10.13039\/501100004442","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["currentprotocols.onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["CP Nucleic Acid Chemistry"],"published-print":{"date-parts":[[2020,9]]},"abstract":"<jats:title>Abstract<\/jats:title><jats:p>RNAs with 5\u2032 functional groups have been gaining interest as molecular probes and reporter molecules. Copper\u2010catalyzed azide\u2010alkyne cycloaddition is one of the most straightforward methods to access such molecules; however, RNA functionalization with azide group has been posing a synthetic challenge. This article describes a simple and efficient protocol for azide functionalization of oligoribonucleotides 5\u2032\u2010end in solid\u2010phase. An azide moiety is attached directly to the C5\u2032\u2010end in two steps: (i) \u2010OH to \u2010I conversion using methyltriphenoxyphosphonium iodide, and (ii) \u2010I to \u2010N<jats:sub>3<\/jats:sub> substitution using sodium azide. The reactivity of the resulting compounds is exemplified by fluorescent labeling using both copper(I)\u2010catalyzed (CuAAC) and strain\u2010promoted (SPAAC) azide\u2010alkyne cycloaddition reactions, ligation of two RNA fragments, and cyclization of short bifunctionalized oligonucleotides. The protocol makes use of oligoribonucleotides synthesized by standard phosphoramidite approach on solid support, using commercially available 2\u2032\u2010<jats:italic>O<\/jats:italic>\u2010PivOM\u2010protected monomers. Such a protection strategy eliminates the interference between the iodination reagent and silyl protecting groups (TBDMS, TOM) commonly used in RNA synthesis by phosphoramidite approach. \u00a9 2020 Wiley Periodicals LLC.<\/jats:p><jats:p><jats:bold>Basic Protocol 1<\/jats:bold>: Solid\u2010phase synthesis of oligoribonucleotide 5\u2032\u2010azides<\/jats:p><jats:p><jats:bold>Basic Protocol 2<\/jats:bold>: CuAAC labeling of oligoribonucleotide 5\u2032\u2010azides in solution<\/jats:p><jats:p><jats:bold>Alternate Protocol 1<\/jats:bold>: CuAAC labeling of oligoribonucleotide 5\u2032\u2010azides on solid support<\/jats:p><jats:p><jats:bold>Basic Protocol 3<\/jats:bold>: SPAAC labeling of oligoribonucleotide 5\u2032\u2010azides<\/jats:p><jats:p><jats:bold>Basic Protocol 4<\/jats:bold>: CuAAC ligation of oligoribonucleotide 5\u2032\u2010azides<\/jats:p><jats:p><jats:bold>Basic Protocol 5<\/jats:bold>: CuAAC cyclization of oligoribonucleotide 5\u2032\u2010azides<\/jats:p><jats:p><jats:bold>Support Protocol<\/jats:bold>: HPLC Purification<\/jats:p>","DOI":"10.1002\/cpnc.112","type":"journal-article","created":{"date-parts":[[2020,7,27]],"date-time":"2020-07-27T18:36:57Z","timestamp":1595875017000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":8,"title":["Solid\u2010Phase Synthesis of RNA 5\u2032\u2010Azides and Their Application for Labeling, Ligation, and Cyclization Via Click Chemistry"],"prefix":"10.1002","volume":"82","author":[{"given":"Marcin","family":"Warminski","sequence":"first","affiliation":[{"name":"Division of Biophysics, Institute of Experimental Physics, Faculty of Physics University of Warsaw  Warsaw Poland"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Joanna","family":"Kowalska","sequence":"additional","affiliation":[{"name":"Division of Biophysics, Institute of Experimental Physics, Faculty of Physics University of Warsaw  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