Chromatin Immunoprecipitation Assay to Analyze the Effect of G-Quadruplex Interactive Agents on the Binding of RNA Polymerase II and Transcription Factors to a Target Promoter Region.

Abstract

Growing evidence suggests the existence of G-quadruplexes and their involvement in transcriptional regulation of many human genes, including VEGF. These studies also provide strong evidence that G-quadruplex structures are stabilized by binding to small molecules, resulting in the modulation of the transcription of genes whose promoters form G-quadruplexes. Here, we describe a chromatin immunoprecipitation (ChIP) assay to determine whether G-quadruplex-interactive agents influence the recruitment of cellular transcription factors, such as Sp1, nucleolin, or hnRNP-K to target genes that contain potential G-quadruplex (G4)-forming sequences in their promoters, subsequently modulating the occupancy of RNA Pol II on the same promoter region.