Temporal Labeling of Nascent RNA Using Photoclick Chemistry in Live Cells

2017/05/31 by Sarah Nainar, Miles Kubota, Christopher McNitt +4 · 19 citations
Chemistry · Materials Science · Biochemistry, Genetics and Molecular Biology · #Click Chemistry and Applications #Photochromic and Fluorescence Chemistry #Advanced biosensing and bioanalysis techniques

paper · doi:10.1021/jacs.7b03121

Abstract

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′N 3 -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.

Citations

• A Strain-Promoted [3 + 2] Azide−Alkyne Cycloaddition for Covalent Modification of Biomolecules in Living Systems
• Selective Labeling of Living Cells by a Photo-Triggered Click Reaction

Cited by

• Multiphoton Activation of Photo-Strain-Promoted Azide Alkyne Cycloaddition “Click” Reagents Enables in Situ Labeling with Submicrometer Resolution
• Light-Triggered Click Chemistry
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• Photo-induced and Rapid Labeling of Tetrazine-Bearing Proteins via Cyclopropenone-Caged Bicyclononynes. [europepmc]
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• Reactive oxygen species-activated bioorthogonal chemistry in living systems enabled by boronate-caged dihydrotetrazines. [europepmc]
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