Quantifying the activity of cis-regulatory elements in the mouse retina by explant electroporation

Methods Mol Biol. 2013:935:329-40. doi: 10.1007/978-1-62703-080-9_23.

Abstract

Transcription factors control gene expression by binding to noncoding regions of DNA known as -cis-regulatory elements (CREs; i.e., enhancer/promoters). Traditionally, cis-regulatory analysis has been carried out via mouse transgenesis which is time-consuming and nonquantitative. Electroporation of DNA reporter constructs into living mouse tissue is a rapid and effective alternative to transgenesis which permits quantitative assessment of cis-regulatory activity. Here, we present a simple technique for quantifying the activity of photoreceptor-specific CREs in living explanted mouse retinas.

MeSH terms

  • Animals
  • DNA / administration & dosage*
  • DNA / genetics
  • Dissection / methods
  • Electroporation / methods*
  • Genes, Reporter
  • Mice
  • Microscopy, Fluorescence / methods
  • Regulatory Sequences, Nucleic Acid*
  • Retina / metabolism*
  • Tissue Culture Techniques / methods
  • Tissue Fixation / methods

Substances

  • DNA