Identifying transcriptional regulatory regions using reporter genes and DNA-protein interactions by chromatin immunoprecipitation

Authors

Lezanne Ooi, University of WollongongFollow
Ian C. Wood, University of LeedsFollow

RIS ID

72640

Publication Details

Ooi, L. & Wood, I. C. (2008). Identifying transcriptional regulatory regions using reporter genes and DNA-protein interactions by chromatin immunoprecipitation. Methods in Molecular Biology, 491 3-17.

Abstract

A comprehensive understanding of regulatory protein interactions with their target genes is fundamental to determining transcriptional networks and identifying important events in the regulation of gene expression. Here we describe how transcriptional regulatory regions are to be identified using luciferase assays (including the transfection of cells by Amaxa and lipid-based reagents) and how protein-DNA interactions are to be characterised by chromatin immunoprecipitation (ChIP) coupled with quantitative PCR. Together these techniques provide a powerful combination for investigating potassium channel gene regulation.