Methods to Discover Alternative Promoter Usage and Transcriptional Regulation of Murine Bcrp1

J Vis Exp. 2016 May 27:(111):53827. doi: 10.3791/53827.

Abstract

Gene expression in different tissues is often controlled by alternative promoters that result in the synthesis of mRNA with unique - usually untranslated - first exons. Bcrp1 (Abcg2), the murine orthologue of the ABC transporter Breast Cancer Resistance Protein (BCRP, ABCG2), has at least four alternative promoters that are designated by the corresponding four alternative first exons produced: E1U, E1A, E1B, and E1C. Herein, in-silico protocols are presented to predict alternative promoter usage for Bcrp1. Furthermore, reporter assay methods are described to produce reporter constructs for alternative promoters and to determine the functionality of putative promoters upstream of the alternative first exons that are identified.

Publication types

  • Video-Audio Media

MeSH terms

  • ATP Binding Cassette Transporter, Subfamily G, Member 2 / biosynthesis
  • ATP Binding Cassette Transporter, Subfamily G, Member 2 / genetics*
  • Animals
  • Base Sequence
  • Exons
  • Gene Expression Regulation
  • Humans
  • Mice
  • Promoter Regions, Genetic*
  • RNA, Messenger / biosynthesis
  • RNA, Messenger / genetics

Substances

  • ATP Binding Cassette Transporter, Subfamily G, Member 2
  • Abcg2 protein, mouse
  • RNA, Messenger