Conditional human VEGF-mediated vascularization in chicken embryos using a novel temperature-inducible gene regulation (TIGR) system

Nucleic Acids Res. 2003 Jun 15;31(12):e69. doi: 10.1093/nar/gng069.


Advanced heterologous transcription control systems for adjusting desired transgene expression are essential for gene function assignments, drug discovery, manufacturing of difficult to produce protein pharmaceuticals and precise dosing of gene-based therapeutic interventions. Conversion of the Streptomyces albus heat shock response regulator (RheA) into an artificial eukaryotic transcription factor resulted in a vertebrate thermosensor (CTA; cold-inducible transactivator), which is able to adjust transcription initiation from chimeric target promoters (P(CTA)) in a low-temperature- inducible manner. Evaluation of the temperature-dependent CTA-P(CTA) interaction using a tailored ELISA-like cell-free assay correlated increased affinity of CTA for P(CTA) with temperature downshift. The temperature-inducible gene regulation (TIGR) system enabled tight repression in the chicken bursal B-cell line DT40 at 41 degrees C as well as precise titration of model product proteins up to maximum expression at or below 37 degrees C. Implantation of microencapsulated DT40 cells engineered for TIGR-controlled expression of the human vascular endothelial growth factor A (hVEGF121) provided low-temperature-induced VEGF-mediated vascularization in chicken embryos.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Bacterial Proteins*
  • Blood Vessels / anatomy & histology
  • Blood Vessels / growth & development
  • Cell-Free System
  • Chick Embryo
  • Cold Temperature*
  • Endothelial Growth Factors / genetics*
  • Endothelial Growth Factors / metabolism
  • Humans
  • Intercellular Signaling Peptides and Proteins / genetics*
  • Intercellular Signaling Peptides and Proteins / metabolism
  • Lymphokines / genetics*
  • Lymphokines / metabolism
  • Neovascularization, Physiologic*
  • Promoter Regions, Genetic
  • Protein Engineering*
  • Repressor Proteins / genetics
  • Repressor Proteins / metabolism
  • Temperature
  • Trans-Activators / genetics
  • Trans-Activators / metabolism
  • Transcriptional Activation*
  • Transfection
  • Transgenes
  • Tumor Cells, Cultured
  • Vascular Endothelial Growth Factor A
  • Vascular Endothelial Growth Factors


  • Bacterial Proteins
  • Endothelial Growth Factors
  • Intercellular Signaling Peptides and Proteins
  • Lymphokines
  • Repressor Proteins
  • RheA protein, Streptomyces albus
  • Trans-Activators
  • Vascular Endothelial Growth Factor A
  • Vascular Endothelial Growth Factors