Temporal Labeling of Nascent RNA Using Photoclick Chemistry in Live Cells

2017/05/31 by Sarah Nainar, Miles Kubota, Christopher McNitt +4 · 67 citations
Biochemistry, Genetics and Molecular Biology · Chemistry · Materials Science · #Advanced biosensing and bioanalysis techniques #Biochemistry #Biophysics #Cell biology #Chemistry #Click Chemistry and Applications #Gene #Nanotechnology #Photochromic and Fluorescence Chemistry #RNA

paper · doi:10.1021/jacs.7b03121

published in Journal of the American Chemical Society 139(24), 8090-8093 (American Chemical Society)

openalex publication_date 2017/05/31 · openalex created_date 2025/10/10 · openalex updated_date 2026/07/26

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
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• Photo‐induced and Rapid Labeling of Tetrazine‐Bearing Proteins via Cyclopropenone‐Caged Bicyclononynes
• Ring Expansion of Alkylidenecarbenes Derived from Lactams, Lactones, and Thiolactones into Strained Heterocyclic Alkynes: A Theoretical Study. [europepmc]
• Expanding the Scope of RNA Metabolic Labeling with Vinyl Nucleosides and Inverse Electron-Demand Diels-Alder Chemistry. [europepmc]
• Photo-induced and Rapid Labeling of Tetrazine-Bearing Proteins via Cyclopropenone-Caged Bicyclononynes. [europepmc]
• Exploring the Condensation Reaction between Aromatic Nitriles and Amino Thiols To Optimize In Situ Nanoparticle Formation for the Imaging of Proteases and Glycosidases in Cells. [europepmc]
• Reduction Triggered In Situ Polymerization in Living Mice. [europepmc]
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• Label-retention expansion microscopy. [europepmc]
• Bioorthogonal chemistry. [europepmc]
• Photoclick Reaction Constructs Glutathione-Responsive Theranostic System for Anti-Tuberculosis. [europepmc]
• The Efficiency of Metabolic Labeling of DNA by Diels-Alder Reactions with Inverse Electron Demand: Correlation with the Size of Modified 2'-Deoxyuridines. [europepmc]
• Precise Regulations at the Subcellular Level through Intracellular Polymerization, Assembly, and Transformation. [europepmc]
• Reactive oxygen species-activated bioorthogonal chemistry in living systems enabled by boronate-caged dihydrotetrazines. [europepmc]
• TagC-RED: An Infrared-Triggered Retro-Ene Reaction for Deep-Tissue Bioconjugation. [europepmc]

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