title: Current covalent modification methods for detecting RNA in fixed and living cells.
abstract: Labeling RNAs is of particular interest for elucidating localization, transport, and regulation of specific transcripts, ideally in living cells. Numerous methods have been developed ranging from hybridizing probes to genetically encoded reporters and chemo-enzymatic approaches. This review focuses on covalent labeling approaches that rely on the introduction of a small reactive group into the nascent or completed transcript followed by bioorthogonal click chemistry. State of the approaches for labeling RNA in fixed and living cells will be presented and emerging strategies with great potential for application in the complex cellular environment will be discussed.
authors.affiliation: Westfälische Wilhelms-Universität Münster, Institute of Biochemistry, 48149 Muenster, Germany.
authors.forename: Josephin M
authors.initials: JM
authors.lastname: Holstein
authors.affiliation: Westfälische Wilhelms-Universität Münster, Institute of Biochemistry, 48149 Muenster, Germany; Cells-in-Motion Cluster of Excellence (EXC 1003 - CiM), University of Muenster, 48149 Muenster, Germany. Electronic address: a.rentmeister@uni-muenster.de.
authors.forename: Andrea
authors.initials: A
authors.lastname: Rentmeister
mesh_terms: D020029:Base Pairing; D002460:Cell Line; D060833:Cellular Microenvironment; D057930:Click Chemistry / Q000379:methods*; D005456:Fluorescent Dyes / Q000737:chemistry; D006801:Humans; D035683:MicroRNAs / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D015335:Molecular Probes / Q000737:chemistry*; D009841:Oligonucleotides / Q000138:chemical synthesis / Q000737:chemistry; D012333:RNA, Messenger / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D000072760:Single Molecule Imaging / Q000295:instrumentation / Q000379:methods*; D060327:Solid-Phase Synthesis Techniques / Q000379:methods; D013194:Staining and Labeling / Q000379:methods*; D016707:Tissue Fixation / Q000379:methods; D014158:Transcription, Genetic
chemical_list: D005456:Fluorescent Dyes; D035683:MicroRNAs; D015335:Molecular Probes; D009841:Oligonucleotides; D012333:RNA, Messenger
keywords: Chemo-enzymatic; Click chemistry; Labeling; RNA visualization