title: Diels-Alder cycloadditions on synthetic RNA in mammalian cells.
abstract: Inverse electron demand Diels-Alder cycloadditions are extremely useful tools for orthogonal labeling of biomolecules such as proteins or small molecules in a cellular context. In-cell labeling of dienophile-modified RNA oligonucleotides using Diels-Alder cycloaddition reactions has not been demonstrated before. In this study we report site-specific labeling of RNA oligonucleotides modified with norbornene derivatives at a predefined sequence position within an RNA sequence in vitro and in mammalian cells using various tetrazine-fluorophore conjugates. The approach could in future be used as a chemical tool for the detection and investigation of RNA functions in cells minimizing the presumed distortion of RNA functions by a large chemical reporter group such as a fluorophore.
authors: LIMES Institute Chemical Biology and Medicinal Chemistry Unit, University of Bonn , Gerhard-Domagk-Strasse 1, 53121 Bonn, Germany.
authors: Anna Maria
authors: AM
authors: Pyka
authors: Christof
authors: C
authors: Domnick
authors: Feodor
authors: F
authors: Braun
authors: Stephanie
authors: S
authors: Kath-Schorr
mesh_terms: D001483:Base Sequence; D002460:Cell Line; D045744:Cell Line, Tumor; D061565:Cycloaddition Reaction; D006573:Heterocyclic Compounds, 1-Ring / Q000737:chemistry; D006801:Humans; D009636:Norbornanes / Q000737:chemistry; D009843:Oligoribonucleotides / Q000138:chemical synthesis / Q000737:chemistry / Q000235:genetics; D012313:RNA / Q000138:chemical synthesis / Q000737:chemistry* / Q000235:genetics; D013194:Staining and Labeling
chemical_list: D006573:Heterocyclic Compounds, 1-Ring; D009636:Norbornanes; D009843:Oligoribonucleotides; D012313:RNA