RILES, a novel method for temporal analysis of the in vivo regulation of miRNA expression. - Université de Tours Accéder directement au contenu
Article Dans Une Revue Nucleic Acids Research Année : 2013

RILES, a novel method for temporal analysis of the in vivo regulation of miRNA expression.

Résumé

Novel methods are required to investigate the complexity of microRNA (miRNA) biology and particularly their dynamic regulation under physiopathological conditions. Herein, a novel plasmid-based RNAi-Inducible Luciferase Expression System (RILES) was engineered to monitor the activity of endogenous RNAi machinery. When RILES is transfected in a target cell, the miRNA of interest suppresses the expression of a transcriptional repressor and consequently switch-ON the expression of the luciferase reporter gene. Hence, miRNA expression in cells is signed by the emission of bioluminescence signals that can be monitored using standard bioluminescence equipment. We validated this approach by monitoring in mice the expression of myomiRs-133, -206 and -1 in skeletal muscles and miRNA-122 in liver. Bioluminescence experiments demonstrated robust qualitative and quantitative data that correlate with the miRNA expression pattern detected by quantitative RT-PCR (qPCR). We further demonstrated that the regulation of miRNA-206 expression during the development of muscular atrophy is individual-dependent, time-regulated and more complex than the information generated by qPCR. As RILES is simple and versatile, we believe that this methodology will contribute to a better understanding of miRNA biology and could serve as a rationale for the development of a novel generation of regulatable gene expression systems with potential therapeutic applications.
Fichier principal
Vignette du fichier
gkt797.pdf (9.2 Mo) Télécharger le fichier
Origine Fichiers éditeurs autorisés sur une archive ouverte

Dates et versions

hal-00874881 , version 1 (22-05-2024)

Identifiants

Citer

Safia Ezzine, Georges Vassaux, Bruno Pitard, Benoit Barteau, Jean-Marc Malinge, et al.. RILES, a novel method for temporal analysis of the in vivo regulation of miRNA expression.. Nucleic Acids Research, 2013, 41 (20), pp.e192. ⟨10.1093/nar/gkt797⟩. ⟨hal-00874881⟩
81 Consultations
0 Téléchargements

Altmetric

Partager

Gmail Mastodon Facebook X LinkedIn More