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C.–Y. Liu, D. Weng, Y. Hayashi, W.W. Kao; Induction of LacZ Expression in Kera3.2–int–rtTA /tet–O–Cre /Rosa26–LacZ Transgenic Mice . Invest. Ophthalmol. Vis. Sci. 2006;47(13):2327.
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© ARVO (1962-2015); The Authors (2016-present)
To develop a mouse line in which a gene of interest can be removed in keratocytes–specific manner upon induction by doxycycline.
Two transgenes including Kera3.2–int–rtTA and tet–O–Cre were co–microinjected into the fertilized mouse eggs to create transgenic mice. The Kera3.2–int–rtTA /tet–O–Cre transgenic mice were crossed with reporter Rosa26–LacZ mice to obtain Kera3.2–int–rtTA /tet–O–Cre/Rosa26–LacZ triple transgenic mice. The Cre activity was assessed by the detection of whole mount X–gal staining in corneal stroma of triple transgenic mice after administration of doxycycline via drinking water and chow.
We obtained two independent transgenic founder lines in which the Kera3.2–int–rtTA and tet–O–Cre transgenes were co–segregated at the chromosome level. Administration of doxycycline (Dox) to transgenic Kera3.2–int–rtTA /tet–O–Cre mice did not induce expression of LacZ. The Kera3.2–int–rtTA /tet–O–Cre/Rosa26–LacZ triple transgenic mice showed a little bit leaky expression of LacZ in the brain even without Dox but not in the eye. Interestingly, upon Dox induction, expression of LacZ can be detected in the keratocytes and the ganglion cell layer and inner nuclear layer of the retina of the Kera3.2–int–rtTA /tet–O–Cre/Rosa26–LacZ triple transgenic mice.
We have established a Kera3.2–int–rtTA /tet–O–Cre transgenic mouse model which can be used to elucidate functions of gene in corneal and retinal development
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