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Yun-Zheng Le, Meili Zhu; Efficient induction of productive Cre-mediated recombination in the retinal pigment epithelium. Invest. Ophthalmol. Vis. Sci. 2014;55(13):4563.
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To dissect gene functions in adult retinal pigmented epithelium (RPE) cells, we previously generated a tetracycline-inducible RPE-specific Cre mouse line (IOVS 49:1248, 2008). To improve the reproducibility of productive Cre-mediated recombination for inducible gene activation and inactivation in the RPE, we recently investigated the inducibility of Cre recombinase in this mouse line.
Analysis of Cre expression was carried out in double transgenic mice derived from inducible RPE-specific Cre and Cre-activatable lacZ reporter R26R mice. Tetracycline derivative, doxycycline (Dox), was supplied to mice intravitreally or intraperitoanlly. Cre expression/function was measured by enzymatic activity and immunohistochemcial staining for beta-galactosidase.
While intraperitoneal Dox injection induced Cre-mediated recombination in some mice, intravitreal Dox delivery resulted in productive Cre-mediated recombination in at least 60% of RPE cells. In addition, a brief Dox induction did not cause any apparent alteration in retinal integrity.
Our results suggest that productive Cre-mediated recombination can be achieved efficiently in mice induced by intravitreal Dox delivery, with no apparent Cre toxicity. Therefore, our inducible RPE-specific Cre mice are suitable for Cre/lox-based gene activation and inactivation in adult RPE, which is critical to the effectiveness and suitability of this mouse line in long-term studies requiring conditional gene targeting.
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