title: Enzymatic Modification of 5'-Capped RNA and Subsequent Labeling by Click Chemistry.
abstract: The combination of enzymatic modification and bioorthogonal click chemistry provides a powerful approach for site-specific labeling of different classes of biomolecules in vitro and even in cellular environments. Herein, we describe a chemoenzymatic method to site specifically label 5'-capped model mRNAs independent of their sequence. A trimethylguanosine synthase was engineered to introduce alkyne, azido, or 4-vinylbenzyl moieties to the 5'-cap. These functional groups were then used for labeling using typical click reactions, such as the azide-alkyne cycloaddition or the tetrazine ligation.
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.
authors.forename: Daniela
authors.initials: D
authors.lastname: Stummer
authors.affiliation: Westfälische Wilhelms-Universität Münster, Institute of Biochemistry, 48149, Muenster, Germany. a.rentmeister@uni-muenster.de.|Cells-in-Motion Cluster of Excellence (EXC 1003 - CiM), University of Muenster, 48149, Muenster, Germany. a.rentmeister@uni-muenster.de.
authors.forename: Andrea
authors.initials: A
authors.lastname: Rentmeister
mesh_terms: D057930:Click Chemistry / Q000379:methods*; D061565:Cycloaddition Reaction; D005456:Fluorescent Dyes / Q000737:chemistry; D008780:Methyltransferases / Q000235:genetics / Q000378:metabolism*; D015394:Molecular Structure; D015202:Protein Engineering; D012315:RNA Caps / Q000378:metabolism*; D012333:RNA, Messenger / Q000737:chemistry* / Q000378:metabolism
chemical_list: D005456:Fluorescent Dyes; D012315:RNA Caps; D012333:RNA, Messenger; D008780:Methyltransferases; C456271:trimethylguanosine synthase
keywords: AdoMet analog; Cap; Chemoenzymatic; Click chemistry; RNA; RNA labeling; RNA modification