Identification of Response Elements on Promoters Using Site-Directed Mutagenesis and Chromatin Immunoprecipitation

Methods Mol Biol. 2018:1765:43-56. doi: 10.1007/978-1-4939-7765-9_3.

Abstract

Proximal promoters are located upstream of the transcription start sites of genes, and they contain regulatory sequences on which bind different transcription factors for promoting colorectal cancer progression. Here we describe the comprehensive methodology used previously for the identification and functional characterization of MYC-responsive elements in the integrin α1 subunit (ITGA1) gene using a combination of in silico analysis, site-directed mutagenesis, and chromatin immunoprecipitation.

Keywords: ChIP; ITGA1; MYC; Mutagenesis; Promoters; Response element; Transcription factor.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Binding Sites
  • Carcinogenesis / genetics
  • Chromatin Immunoprecipitation / instrumentation
  • Chromatin Immunoprecipitation / methods*
  • Colorectal Neoplasms / genetics*
  • Computer Simulation
  • Gene Expression Regulation, Neoplastic
  • HEK293 Cells
  • Humans
  • Integrin alpha1 / genetics*
  • Mutagenesis, Site-Directed / instrumentation
  • Mutagenesis, Site-Directed / methods*
  • Proto-Oncogene Proteins c-myc / metabolism
  • Response Elements / genetics*
  • Sequence Analysis, DNA / methods
  • Transcription Initiation Site

Substances

  • Integrin alpha1
  • MYC protein, human
  • Proto-Oncogene Proteins c-myc