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L. Feiner, M. Navaratnarajah, E.A. Pierce; Photoreceptor Expression of Cre-Recombinase . Invest. Ophthalmol. Vis. Sci. 2003;44(13):3539.
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Purpose: The use of cre and other recombinases for conditional gene targeting allows removal of genes in a tissuespecific manner. Many genes implicated in ocular disease have been studied using traditional transgenic and knock-out strategies. However, some genes important in disease pathophysiology are critical for the normal development and homeostasis, and thus cannot be studied by these techniques. Conditional gene expression in the eye using cre-recombinase is one technique that will allow genes to be turned on or off in a specfic subset of cells. We have produced transgenic mice that express cre-recombinase in photoreceptors. Methods: The gene encoding cre-recombinase with an N-terminal nuclear localization sequence was placed behind a 5kb fragment from the rhodopsin promoter. Three independent lines of cre transgenic mice were generated and crossed to the ROSA26 line of floxed beta-galactosidase reporter mice. Retinas from doubly transgenic progeny and control littermates were evaluated by X-gal histochemistry in adult and neonatal mice in whole-mount preparation and in frozen sections. Results: All three rhodopsin cre transgenic mouse lines express cre-recombinase in photoreceptors. Activation of beta-galactosidase is present in all rods thoughout the entire retina. Beta-galactosidase activity can bedetected by postnatal day 4. Activation of beta-galactosidase has not been detected in any tissuesoutside the retina. Histologic analysis from retinas of mice from 4 month old animals appears normal and without signs of retinal degeneration. Analysis of retinal function in the transgenic animals by ERG is underway. Conclusions: The rhodopsin promoter is sufficient to drive retinal expression of cre recombinase and activate transcription of functional beta-galactosidase in the retina. Crosses between these rhodopsin cre-transgenic mice and mice with conditionally targeted genes will allow us to investigate the role of these genes in photoreceptor development and function.
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