Temporal Labeling of Nascent RNA Using Photoclick Chemistry in Live Cells,Journal of the American Chemical Society

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Temporal Labeling of Nascent RNA Using Photoclick Chemistry in Live Cells
Journal of the American Chemical Society ( IF 14.4 ) Pub Date : 2017-06-08 , DOI: 10.1021/jacs.7b03121
Sarah Nainar , Miles Kubota , Christopher McNitt ₁ , Catherine Tran , Vladimir V. Popik ₁ , Robert C. Spitale

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We report the first cellular application of a photoclick SPAAC reagent to label azide-functionalized RNA. 350 nm irradiation of a cyclopropenone caged oxo-dibenzocyclooctyne (photo-ODIBO) biotin yields formation of the SPAAC reactive species, which rapidly forms adducts with RNA containing 2'-azidoadenosine (2'N3-A). Photo-ODIBO was found to be highly stable in the presence of thiols, conferring greater stability relative to ODIBO. Light activated photo-ODIBO enabled tagging of cellular RNA, in addition to fluorescent imaging as well as enrichment of RNA in cell subpopulations via selective irradiation.

中文翻译：

在活细胞中使用 Photoclick 化学对新生 RNA 进行时间标记

我们报告了 photoclick SPAAC 试剂的首次细胞应用，用于标记叠氮化物功能化的 RNA。350 nm 辐照环丙烯酮笼状氧代-二苯并环辛炔 (photo-ODIBO) 生物素产生 SPAAC 反应性物质的形成，其与含有 2'-叠氮基腺苷 (2'N3-A) 的 RNA 快速形成加合物。发现 Photo-ODIBO 在硫醇存在下高度稳定，相对于 ODIBO 具有更高的稳定性。除了荧光成像以及通过选择性辐照在细胞亚群中富集 RNA 之外，光激活的 photo-ODIBO 还能够标记细胞 RNA。

更新日期：2017-06-08

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