Sun Daekyu
College of Pharmacy, University of Arizona, Tucson, AZ, USA.
BIO5 Institute, Tucson, AZ, USA.
Methods Mol Biol. 2019;2035:233-242. doi: 10.1007/978-1-4939-9666-7_13.
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.
越来越多的证据表明G-四链体的存在及其参与包括血管内皮生长因子（VEGF）在内的许多人类基因的转录调控。这些研究还提供了有力证据，即G-四链体结构通过与小分子结合而稳定，从而导致其启动子形成G-四链体的基因转录受到调节。在此，我们描述了一种染色质免疫沉淀（ChIP）测定法，以确定G-四链体相互作用剂是否会影响细胞转录因子（如Sp1、核仁素或hnRNP-K）募集到其启动子中含有潜在G-四链体（G4）形成序列的靶基因上，进而调节RNA聚合酶II在同一启动子区域的占据情况。
