The human cGMP-PDE beta-subunit promoter region directs expression of the gene to mouse photoreceptors

Invest Ophthalmol Vis Sci. 2000 Dec;41(13):4059-63.


Purpose: We previously demonstrated that 350 bp of the human rod cGMP phosphodiesterase beta-subunit (beta-PDE) gene promoter are sufficient to direct high levels of gene expression in human Y-79 retinoblastoma cells in vitro. In this study the cell specificity and expression pattern conferred by the short beta-PDE 5' flanking sequence in vivo were examined.

Methods: A construct containing the bacterial LacZ gene driven by a fragment of the beta-PDE 5' flanking region (-297 to +53) was used to generate transgenic mice. Gene expression was analyzed by measuring beta-galactosidase activity in tissue homogenates or visualizing enzymatic activity or protein production at a cellular level by in situ histochemistry or immunocytochemistry.

Results: Three independently derived transgenic lines were generated carrying the -297 to +53 beta-PDE 5' flanking region fragment. Within the retina, the reporter gene was specifically expressed in photoreceptors, consistent with the localization of endogenous beta-PDE. Significant expression of LacZ was not observed in other ocular or peripheral tissues.

Conclusions: Photoreceptor-specific reporter gene expression is driven in vivo by a 350-bp segment of the beta-PDE 5' flanking sequence. This study demonstrates the utility of the human beta-PDE promoter for directing the expression of foreign genes to photoreceptors and suggests that the -297 to +53 beta-PDE 5' flanking region fragment may have important implications for therapeutic gene delivery to the visual cells.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • 3',5'-Cyclic-GMP Phosphodiesterases / biosynthesis
  • 3',5'-Cyclic-GMP Phosphodiesterases / genetics*
  • Animals
  • Cyclic Nucleotide Phosphodiesterases, Type 6
  • DNA Primers / chemistry
  • Fluorescent Antibody Technique, Indirect
  • Gene Expression*
  • Genes, Reporter / genetics
  • Lac Operon / genetics
  • Mice
  • Mice, Inbred C57BL
  • Mice, Transgenic
  • Microscopy, Confocal
  • Phosphoric Diester Hydrolases*
  • Photoreceptor Cells, Vertebrate / enzymology*
  • Promoter Regions, Genetic / genetics*
  • Recombinant Fusion Proteins / genetics
  • Recombinant Fusion Proteins / metabolism
  • Tissue Distribution
  • beta-Galactosidase / genetics
  • beta-Galactosidase / metabolism


  • DNA Primers
  • Recombinant Fusion Proteins
  • Phosphoric Diester Hydrolases
  • 3',5'-Cyclic-GMP Phosphodiesterases
  • Cyclic Nucleotide Phosphodiesterases, Type 6
  • PDE6B protein, human
  • Pde6b protein, mouse
  • beta-Galactosidase