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The Human cGMP-PDE ß-Subunit Promoter Region Directs Expression of the Gene to Mouse Photoreceptors

¹ From the Department of Ophthalmology and Jules Stein Eye Institute, UCLA School of Medicine, Los Angeles, California; ² Molecular Biology Institute, UCLA; ³ Centre for Research in Neuroscience, McGill University, Montreal, Canada; and ⁴ Departments of Vision Science and Neuroscience, University of California, Berkeley, California.

PURPOSE. We previously demonstrated that 350 bp of the human rod cGMP phosphodiesterase ß-subunit (ß-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 ß-PDE 5' flanking sequence in vivo were examined.

METHODS. A construct containing the bacterial LacZ gene driven by a fragment of the ß-PDE 5' flanking region (-297 to +53) was used to generate transgenic mice. Gene expression was analyzed by measuring ß-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 ß-PDE 5' flanking region fragment. Within the retina, the reporter gene was specifically expressed in photoreceptors, consistent with the localization of endogenous ß-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 ß-PDE 5' flanking sequence. This study demonstrates the utility of the human ß-PDE promoter for directing the expression of foreign genes to photoreceptors and suggests that the -297 to +53 ß-PDE 5' flanking region fragment may have important implications for therapeutic gene delivery to the visual cells.