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 demon strated 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 mitt 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