In Vitro Approaches to Identify Thyroid Hormone Receptor-Dependent Transcriptional Response.
Fiche publication
Date publication
janvier 2018
Journal
Methods in molecular biology (Clifton, N.J.)
Auteurs
Membres identifiés du Cancéropôle Est :
Dr PLATEROTI Michelina
Tous les auteurs :
Sirakov M, Plateroti M
Lien Pubmed
Résumé
The thyroid hormones act through their nuclear receptor TRs that are T3-modulated transcription factors. To elucidate the molecular mechanisms at the basis of specific physiological responses to TH-TR, in vitro approaches are commonly used to demonstrate the activation or repression of target genes. These approaches allow identifying direct transcriptional targets of TRs which can eventually be confirmed by using in vivo chromatin binding assays.Here, we describe two classical approaches in vitro and in cell lines such as Electro-Mobility Shift Assay and thyroid-hormone-responsive Luciferase-reporter assay (EMSA and Luc, respectively) we have been largely using to investigate TRα1-driven TH-TRs intestinal epithelium cell autonomous action.
Mots clés
DNA binding, Electro Mobility Shift Assay (EMSA), Luciferase assay (Luc), Target gene promoters, Thyroid hormone Response Element (TRE), Transcriptional activity
Référence
Methods Mol Biol. 2018 ;1801:29-38