Temporal Labeling of Nascent RNA Using Photoclick Chemistry in Live Cells
| Autor: | Catherine Tran, Vladimir V. Popik, Robert C. Spitale, Miles Kubota, Sarah Nainar, Christopher D. McNitt |
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| Rok vydání: | 2017 |
| Předmět: |
Cyclopropanes
Cell Survival 010402 general chemistry Fluorescent imaging 01 natural sciences Biochemistry Catalysis Adduct chemistry.chemical_compound Colloid and Surface Chemistry Biotin Humans Cyclopropenone Molecular Structure 010405 organic chemistry Light activated RNA General Chemistry Photochemical Processes 0104 chemical sciences chemistry Alkynes Reagent Click chemistry Click Chemistry HeLa Cells |
| Zdroj: | Journal of the American Chemical Society. 139:8090-8093 |
| ISSN: | 1520-5126 0002-7863 |
| DOI: | 10.1021/jacs.7b03121 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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