3 years ago

Temporal Labeling of Nascent RNA Using Photoclick Chemistry in Live Cells

Temporal Labeling of Nascent RNA Using Photoclick Chemistry in Live Cells
Robert C. Spitale, Vladimir V. Popik, Christopher McNitt, Sarah Nainar, Miles Kubota, Catherine Tran
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.

Publisher URL: http://dx.doi.org/10.1021/jacs.7b03121

DOI: 10.1021/jacs.7b03121

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