Lentiviral vectors with amplified beta cell-specific gene expression

Gene Ther. 2009 Aug;16(8):998-1008. doi: 10.1038/gt.2009.49. Epub 2009 May 14.


An important goal of gene therapy is to be able to deliver genes, so that they express in a pattern that recapitulates the expression of an endogenous cellular gene. Although tissue-specific promoters confer selectivity, in a vector-based system, their activity may be too weak to mediate detectable levels in gene-expression studies. We have used a two-step transcriptional amplification system to amplify gene expression from lentiviral vectors using the human insulin promoter. In this system, the human insulin promoter drives expression of a potent synthetic transcription activator (the yeast GAL4 DNA-binding domain fused to the activation domain of the Herpes simplex virus-1 VP16 activator), which in turn activates a GAL4-responsive promoter, driving the enhanced green fluorescent protein reporter gene. Vectors carrying the human insulin promoter did not express in non-beta-cell lines, but expressed in murine insulinoma cell lines, indicating that the human insulin promoter was capable of conferring cell specificity of expression. The insulin-amplifiable vector was able to amplify gene expression five to nine times over a standard insulin-promoter vector. In primary human islets, gene expression from the insulin-promoted vectors was coincident with insulin staining. These vectors will be useful in gene-expression studies that require a detectable signal and tissue specificity.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Cell Line
  • Gene Expression
  • Gene Transfer Techniques*
  • Genetic Vectors*
  • Green Fluorescent Proteins / genetics
  • Humans
  • Insulin / genetics*
  • Insulin-Secreting Cells / metabolism*
  • Islets of Langerhans / metabolism
  • Lentivirus / genetics*
  • Mice
  • Organ Specificity
  • Phosphoglycerate Kinase / genetics
  • Promoter Regions, Genetic
  • Transcriptional Activation / genetics*


  • Insulin
  • enhanced green fluorescent protein
  • Green Fluorescent Proteins
  • Phosphoglycerate Kinase