Traditional Chemical Mapping of RNA Structure In Vitro and In Vivo.
Fiche publication
Date publication
janvier 2016
Journal
Methods in molecular biology (Clifton, N.J.)
Auteurs
Membres identifiés du Cancéropôle Est :
Dr ROMBY Pascale
Tous les auteurs :
Fechter P, Parmentier D, Wu Z, Fuchsbauer O, Romby P, Marzi S
Lien Pubmed
Résumé
Chemical probing is often used to gain knowledge on the secondary and tertiary structures of RNA molecules either free or engaged in complexes with ligands. The method monitors the reactivity of each nucleotide towards chemicals of various specificities reflecting the hydrogen bonding environment of each nucleotide within the RNA molecule. In addition, information can be obtained on the binding site of a ligand (noncoding RNAs, protein, metabolites), and on RNA conformational changes that accompanied ligand binding or perturbation of the environmental cues. The detection of the modifications can be obtained either by using end-labeled RNA molecules or by primer extension using reverse transcriptase. The goal of this chapter is to provide the reader with an experimental guide to probe the structure of RNA in vitro and in vivo with the most suitable chemical probes.
Mots clés
Computational Biology, methods, Electrophoresis, Polyacrylamide Gel, Models, Molecular, Nucleic Acid Conformation, RNA, chemistry, RNA Cleavage, RNA Folding
Référence
Methods Mol. Biol.. 2016 ;1490:83-103