Absolute quantification of noncoding RNA by microscale thermophoresis.
Fiche publication
Date publication
mars 2019
Journal
Angewandte Chemie (International ed. in English)
Auteurs
Membres identifiés du Cancéropôle Est :
Pr MOTORINE Iouri, Dr MARCHAND Virginie
Tous les auteurs :
Jacob D, Thüring K, Galliot A, Marchand V, Galvanin A, Ciftci A, Scharmann K, Stock M, Roignant JY, Leidel SA, Motorin Y, Schaffrath R, Klassen R, Helm M
Lien Pubmed
Résumé
Accurate quantification of copy numbers of noncoding RNA has recently emerged as an urgent problem, with impact on fields such as RNA modification research, tissue differentiation, and others. Here we present a hybridization-based approach that uses microscale thermophoresis (MST) as a very fast and highly precise readout to quantify e.g. single tRNA species with a turnaround time of about one hour. We developed MST in to quantify the effect of tRNA toxins and of heat stress and RNA modification. A comparative analysis also revealed significant differences to RNA-Seq based quantification approaches, strongly suggesting a bias due to tRNA modifications in the latter. Further applications included quantification of rRNA, as well as of polyA levels in cellular RNA.
Référence
Angew. Chem. Int. Ed. Engl.. 2019 Mar 20;: