Analysis of post-transcriptional regulation using the FunREG method.
An increasing number of arguments, including altered microRNA expression, support the idea that post-transcriptional deregulation participates in gene disturbances found in diseased tissues. To evaluate this hypothesis, we developed a method which facilitates post-transcriptional investigations in a wide range of human cells and experimental conditions. This method, called FunREG (functional, integrated and quantitative method to measure post-transcriptional regulation), connects lentiviral transduction with a fluorescent reporter system and quantitative PCR. Using FunREG, we efficiently measured post-transcriptional regulation mediated either by selected RNA sequences or regulatory factors (microRNAs), and then evaluated the contribution of mRNA decay and translation efficiency in the observed regulation. We demonstrated the existence of gene-specific post-transcriptional deregulation in liver tumour cells, and also reported a molecular link between a transcript variant abrogating HDAC6 (histone deacetylase 6) regulation by miR-433 and a rare familial genetic disease. Because FunREG is sensitive, quantitative and easy to use, many applications can be envisioned in fundamental and pathophysiological research.
INSERM, U889, Groupe de Recherche pour l'Etude du Foie (GREF), Université Victor Sagalen Bordeaux 2, Bordeaux cedex, F-33076 France.
Benoît
B
Laloo
Marion
M
Maurel
Sandra
S
Jalvy-Delvaille
Francis
F
Sagliocco
Christophe F
CF
Grosset
D001483:Base Sequence; D005786:Gene Expression Regulation; D017930:Genes, Reporter; D006801:Humans; D008113:Liver Neoplasms / Q000235:genetics / Q000378:metabolism; D016133:Polymerase Chain Reaction / Q000379:methods*; D012323:RNA Processing, Post-Transcriptional; D020871:RNA Stability / Q000235:genetics; D019076:Transgenes