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Dissection of a Mouse Eye for a Whole Mount of the Retinal Pigment Epithelium
Published on: February 27, 2011
Temporally controlled targeted somatic mutagenesis in mouse eye pigment epithelium
Mikiro Mori1, Laetitia Gargowitsch, Jean-Marc Bornert
1Institut de Génétique et de Biologie Moléculaire et Cellulaire (IGBMC), CNRS UMR7104/INSERM U964, Collège de France, Université de Strasbourg, Illkirch Cedex, France.
Abstract:
To generate temporally controlled site-specific somatic mutations in the mouse eye pigment epithelium, we generated a TRP1-Cre-ER(T2) transgenic mouse line that expresses the tamoxifen-dependent Cre-ER(T2) recombinase under the control of the tyrosinase-related protein 1 (TRP1) promoter. Cre-ER(T2) transcripts were readily detected in the retinal pigment epithelium (RPE), and tamoxifen treatment of adult TRP1-Cre-ER(T2) transgenic mice induced efficient excision of floxed DNA in patches of RPE cells, in numerous epithelial cells of the iris and ciliary body, and in very few cells of the neural retina. Importantly, no excision was detected in any cells in the absence of tamoxifen treatment. Thus, the TRP1-Cre-ER(T2) mouse line provides a powerful tool to study in vivo gene functions in the mouse eye pigment epithelium.
Insights
A new TRP1-Cre-ER(T2) mouse line enables precise genetic modification in the eye's pigment epithelium. This tool allows researchers to study gene functions in vivo within specific retinal pigment epithelial cells.
Area of Science:
- Ophthalmology
- Genetics
- Molecular Biology
Background:
- Development of genetically engineered mouse models is crucial for understanding complex biological processes.
- Site-specific somatic mutations are essential for studying gene function in a temporal and spatial manner within specific cell populations.
- The retinal pigment epithelium (RPE) plays a vital role in ocular health and disease, necessitating tools for its targeted genetic manipulation.
Discussion:
- The study introduces a novel TRP1-Cre-ER(T2) transgenic mouse line for inducible gene manipulation in the eye.
- Tamoxifen administration in adult mice resulted in efficient, site-specific DNA excision in RPE and ocular pigment cells.
- The TRP1 promoter drives Cre-ER(T2) expression effectively in the RPE, enabling temporally controlled genetic alterations.
Key Insights:
- The TRP1-Cre-ER(T2) mouse line provides a robust system for generating temporally controlled, site-specific somatic mutations in the mouse eye pigment epithelium.
- Tamoxifen-inducible Cre-ER(T2) recombinase activity is confirmed in the RPE, iris, and ciliary body, with minimal activity in the neural retina.
- The absence of recombination without tamoxifen highlights the system's specificity and safety for in vivo studies.
Outlook:
- This mouse model offers a powerful platform for investigating gene function and developing therapeutic strategies for RPE-related diseases.
- Future research can leverage this tool to explore the roles of specific genes in RPE development, maintenance, and disease pathogenesis.
- The TRP1-Cre-ER(T2) line facilitates detailed studies of ocular pigment biology and associated disorders.

