Acquisition of Temporal HIF Transcriptional Activity Using a Secreted Luciferase Assay

Methods Mol Biol. 2018:1742:37-44. doi: 10.1007/978-1-4939-7665-2_4.

Abstract

Here we describe a simple method based on secreted luciferase driven by a hypoxia-inducible factor (HIF) response element (HRE) that allows the acquisition of dynamic and high-throughput data on HIF transcriptional activity during hypoxia and pharmacological activation of HIF. The sensitivity of the assay allows for the secreted luciferase to be consecutively sampled (as little as 1% of the total supernatant) over an extended time period, thus allowing the acquisition of time-resolved HIF transcriptional activity.

Keywords: Activity; HIF; Hypoxia; Luciferase; Time-resolved; Transcription.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acids, Dicarboxylic / metabolism
  • Cell Hypoxia
  • HEK293 Cells
  • Humans
  • Hypoxia-Inducible Factor 1, alpha Subunit / genetics*
  • Luciferases / genetics*
  • Promoter Regions, Genetic
  • Recombinant Proteins
  • Response Elements*
  • Transcriptional Activation

Substances

  • Amino Acids, Dicarboxylic
  • HIF1A protein, human
  • Hypoxia-Inducible Factor 1, alpha Subunit
  • Recombinant Proteins
  • Luciferases
  • oxalylglycine