Monitoring of transcriptional dynamics of HIF and NFκB activities

Methods Mol Biol. 2014:1098:97-105. doi: 10.1007/978-1-62703-718-1_8.

Abstract

Genetic experiments over the last few decades have identified many regulatory proteins critical for DNA transcription. The dynamics of their transcriptional activities shape the differential expression of the genes they control. Here we describe a simple method, based on the secreted luciferase, to measure the activities of two transcription factors NFκB and HIF. This technique can effectively monitor dynamics of transcriptional events in a population of cells and be up-scaled for high-throughput screening and promoter analysis, making it ideal for data-demanding applications such as mathematical modelling.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Biosensing Techniques / methods*
  • Copepoda / enzymology
  • Genes, Reporter / genetics
  • HEK293 Cells
  • Humans
  • Hypoxia-Inducible Factor 1 / metabolism*
  • Luciferases / genetics
  • NF-kappa B / metabolism*
  • Signal Transduction
  • Transcription, Genetic*
  • Transfection

Substances

  • Hypoxia-Inducible Factor 1
  • NF-kappa B
  • Luciferases