Bio-orthogonal chemistry-based method for fluorescent labelling of ribosomal RNA in live mammalian cells
| Autor: | Muhan He, P. N. Asare Okai, Qishan Lin, Maksim Royzen, Kui Wu, Irfan Khan, G. Fuchs |
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| Rok vydání: | 2019 |
| Předmět: |
Fluorophore
Conjugated system 010402 general chemistry 01 natural sciences Catalysis Tetrazine chemistry.chemical_compound Tandem Mass Spectrometry Materials Chemistry Humans Chromatography High Pressure Liquid Fluorescent Dyes Cycloaddition Reaction 010405 organic chemistry Metals and Alloys Phosphoramidate General Chemistry Ribosomal RNA Fluorescence 0104 chemical sciences Surfaces Coatings and Films Electronic Optical and Magnetic Materials chemistry Biochemistry RNA Ribosomal Agarose gel electrophoresis Ceramics and Composites Nucleic acid HeLa Cells |
| Zdroj: | Chemical Communications. 55:10456-10459 |
| ISSN: | 1364-548X 1359-7345 |
| DOI: | 10.1039/c9cc05346h |
| Popis: | A bio-orthogonal chemistry-based approach for fluorescent labelling of ribosomal RNA is described. It involves an adenosine analogue modified with trans-cyclooctene and masked 5'-phosphate group using aryl phosphoramidate. The incorporation into rRNA has been confirmed using agarose gel electrophoresis, as well as a highly sensitive UHPLC-MS/MS method. Fluorescent labelling of rRNA has been achieved in live HeLa cells via an inverse electron demand Diels-Alder reaction with a tetrazine conjugated to an Oregon Green fluorophore. This communication describes the stepwise approach that led to the development and characterization of the probe. The results demonstrate a new strategy towards development of future fluorescent probes to investigate the biochemistry of nucleic acids. |
| Databáze: | OpenAIRE |
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