PURPOSE: Generate site-specific somatic mutations selectively in the retinal pigment epithelium (RPE) in mice. METHODS: A transgenic mouse line expressing the Cre recombinase under the control of the tyrosinase-related protein (TRP)-1 promoter was generated. The presence of Cre was determined by in situ hybridization, and Cre-mediated excision of DNA was analyzed by PCR and alkaline phosphatase (AP) histochemistry in reporter mice carrying a loxP-flanked (floxed) retinoid X receptor alpha (RXRa) gene and in Z/AP mice, respectively. RESULTS: Cre was expressed in the RPE from embryonic day 10.5 to postnatal day 12, resulting in efficient floxed excision of DNA in the RPE from embryonic day 10.5 to adulthood in TRP1-Cre mice. Expressed Cre and excision of DNA were also detected in the ciliary margin of the retina and in some cells in the neural retina, but not in the embryonic periocular mesenchyme or in the choroid. CONCLUSIONS: The TRP1-Cre mouse line, which induces efficient Cre-mediated excision of DNA selectively in the RPE, provides a new, powerful tool to study gene functions in the RPE in vivo.
PURPOSE: Generate site-specific somatic mutations selectively in the retinal pigment epithelium (RPE) in mice. METHODS: A transgenic mouse line expressing the Cre recombinase under the control of the tyrosinase-related protein (TRP)-1 promoter was generated. The presence of Cre was determined by in situ hybridization, and Cre-mediated excision of DNA was analyzed by PCR and alkaline phosphatase (AP) histochemistry in reporter mice carrying a loxP-flanked (floxed) retinoid X receptor alpha (RXRa) gene and in Z/AP mice, respectively. RESULTS: Cre was expressed in the RPE from embryonic day 10.5 to postnatal day 12, resulting in efficient floxed excision of DNA in the RPE from embryonic day 10.5 to adulthood in TRP1-Cre mice. Expressed Cre and excision of DNA were also detected in the ciliary margin of the retina and in some cells in the neural retina, but not in the embryonic periocular mesenchyme or in the choroid. CONCLUSIONS: The TRP1-Cre mouse line, which induces efficient Cre-mediated excision of DNA selectively in the RPE, provides a new, powerful tool to study gene functions in the RPE in vivo.
Authors: Aristomenis Thanos; Yuki Morizane; Yusuke Murakami; Andrea Giani; Dimosthenis Mantopoulos; Maki Kayama; Mi In Roh; Norman Michaud; Basil Pawlyk; Michael Sandberg; Lucy H Young; Joan W Miller; Demetrios G Vavvas Journal: Am J Pathol Date: 2012-03-17 Impact factor: 4.307
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Authors: Laura Fernández-Sánchez; Luis Pérez de Sevilla Müller; Nicholas C Brecha; Nicolás Cuenca Journal: Invest Ophthalmol Vis Sci Date: 2014-08-12 Impact factor: 4.799
Authors: Tal Burstyn-Cohen; Erin D Lew; Paqui G Través; Patrick G Burrola; Joseph C Hash; Greg Lemke Journal: Neuron Date: 2012-12-20 Impact factor: 17.173
Authors: Yi Zhou; Christopher Tanzie; Zhipeng Yan; Shuyi Chen; Michael Duncan; Karin Gaudenz; Hua Li; Christopher Seidel; Brandy Lewis; Andrea Moran; Richard T Libby; Amy E Kiernan; Ting Xie Journal: Proc Natl Acad Sci U S A Date: 2013-05-15 Impact factor: 11.205