Identifying transcriptional regulatory regions using reporter genes and DNA-protein interactions by chromatin immunoprecipitation.
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.
Ooi
Lezanne
L
Institute of Membrane and Systems Biology, University of Leeds, UK.
Wood
Ian C
IC
D000818:Animals; D047369:Chromatin Immunoprecipitation / Q000379:methods*; D018274:Electroporation; D018507:Gene Expression Regulation, Developmental; D017930:Genes, Reporter; D006801:Humans; D008156:Luciferases / Q000235:genetics; D020411:Oligonucleotide Array Sequence Analysis; D010957:Plasmids; D016133:Polymerase Chain Reaction; D015221:Potassium Channels / Q000235:genetics*; D011401:Promoter Regions, Genetic; D012045:Regulatory Sequences, Nucleic Acid; D012501:Saphenous Vein / Q000502:physiology; D014158:Transcription, Genetic; D014162:Transfection
D015221:Potassium Channels; D008156:Luciferases