Site-specific somatic mutagenesis in the retinal pigment epithelium.
OBJECTIVE
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.
Mori
Mikiro
M
Institute of Genetics and Molecular and Cellular Biology (IGBMC), CNRS/INSERM/ULP/Collège de France, BP 163, 67404 Illkirch Cedex, CU de Strasbourg, France.
Metzger
Daniel
D
Garnier
Jean-Marie
JM
Chambon
Pierre
P
Mark
Manuel
M
D000818:Animals; D004247:DNA / Q000378:metabolism; D004622:Embryo, Mammalian; D005260:Female; D006822:Hybrid Cells; D017403:In Situ Hybridization; D019426:Integrases / Q000235:genetics*; D008297:Male; D008562:Membrane Glycoproteins; D051379:Mice; D008822:Mice, Transgenic; D016297:Mutagenesis, Site-Directed; D010088:Oxidoreductases; D010857:Pigment Epithelium of Eye / Q000201:enzymology*; D016133:Polymerase Chain Reaction; D011506:Proteins / Q000235:genetics*; D018168:Receptors, Retinoic Acid / Q000235:genetics; D011993:Recombinant Fusion Proteins / Q000235:genetics; D047488:Retinoid X Receptors; D014157:Transcription Factors / Q000235:genetics; D014764:Viral Proteins / Q000235:genetics*
D008562:Membrane Glycoproteins; D011506:Proteins; D018168:Receptors, Retinoic Acid; D011993:Recombinant Fusion Proteins; D047488:Retinoid X Receptors; D014157:Transcription Factors; D014764:Viral Proteins; D004247:DNA; D010088:Oxidoreductases; C079621:Tyrp1 protein, mouse; C064056:tyrosinase-related protein-1; C045073:Cre recombinase; D019426:Integrases