Accéder au contenu
Merck

PCR-based profiling of transcription factor activity in vivo by a virus-based reporter battery.

iScience (2022-03-15)
Hitomi Abe, Kentaro Abe
RÉSUMÉ

Understanding the molecular mechanisms of gene regulation is pivotal for understanding how cells establish and modify their identities and functions. Multiple transcription factors (TFs) coordinate to alter gene expression in cells; however, a method to quantitatively analyze the activity of each TF is lacking, particularly in vivo. Here, we introduce a viral-vector-based TF reporter battery that can be used to simultaneously analyze the activity of multiple TFs, visualized as the TF activity profile (TFAP) obtained by qPCR. We show that the cells possess distinct TFAPs that dynamically change according to experimental manipulation or physiological activity. We report a practical method to obtain the TFAP of a defined cell population and their experience-dependent changes in the mouse brain in vivo. The TFAP obtained by our method will help bridge the information gap between the genome and transcriptome and aid the multi-omics view of understanding the gene regulation system.

MATÉRIAUX
Référence du produit
Marque
Description du produit

Sigma-Aldrich
Hydrocortisone, BioReagent, suitable for cell culture
Sigma-Aldrich
Anticorps anti-NeuN, clone A60, clone A60, Chemicon®, from mouse
Millipore
ANTI-FLAG® antibody produced in rabbit, affinity isolated antibody, buffered aqueous solution
Sigma-Aldrich
Sous-unité B de la toxine cholérique, ≥95% (SDS-PAGE), lyophilized powder
Sigma-Aldrich
D-(–)-2-Amino-5-phosphonopentanoic Acid, Active enantiomer of DL-2-amino-5-phosphonopentanoic acid (AP5) that is a commonly used as a competitive NMDA receptor antagonist.