In silico Analysis and Site-directed Mutagenesis of Promoters

Salah Boudjadi; Jean-Francois Beaulieu

doi:10.21769/BioProtoc.2181

In silico Analysis and Site-directed Mutagenesis of Promoters

Bio Protoc. 2017 Mar 20;7(6):2181. doi: 10.21769/BioProtoc.2181.

Authors

Salah Boudjadi¹, Jean-Francois Beaulieu²

Affiliations

¹ Cancer Molecular Pathology Section, Laboratory of Pathology, National Cancer Institute, National Institutes of Health, Bethesda, USA.
² Laboratory of Intestinal Physiopathology, Department of Anatomy and Cell Biology, Faculty of Medicine and Health Sciences, Université de Sherbrooke, Sherbrooke, Canada.

Abstract

In normal as in cancerous cells, gene expression is tightly regulated by transcription factors, which are responsible for up- or down-regulation of thousands of targets involved in different cell processes. Transcription factors can directly regulate the expression of genes by binding to specific DNA sequences known as response elements. Identification of these response elements is important to characterize targets of transcription factors in order to understand their contribution to gene regulation. Here, we describe in silico analysis coupled to selected mutagenesis and promoter gene reporter assay procedures to identify and analyze response elements in the proximal promoter sequence of genes.

Keywords: Gene expression; Promoter gene-reporter assay; Response elements; Site-directed mutagenesis; Transcription factors; in silico analysis.