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An efficient inducible RPE-Selective cre transgenic mouse line.
- Source :
-
Experimental eye research [Exp Eye Res] 2021 Jan; Vol. 202, pp. 108370. Date of Electronic Publication: 2020 Nov 29. - Publication Year :
- 2021
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Abstract
- Cre-mediated modulation of gene function in the murine retinal pigment epithelium (RPE) has been widely used, but current postnatal RPE-selective Cre driver lines suffer from limited recombination efficiency and/or ectopic or mosaic expression. We sought to generate a transgenic mouse line with consistently efficient RPE-selective Cre activity that could be temporally regulated. We used ϕC31 integrase to insert a DNA construct encoding a human BEST1 promoter fragment driving a Cre recombinase estrogen receptor fusion (BEST1-CreERT2) at the Rosa26 locus of C57BL/6J mice. Rosa26 <superscript>BEST1-CreERT2</superscript> mice were bred with a tdTomato reporter line and to mice with a Cre-conditional allele of Tfam. 4-hydroxytamoxifen or vehicle was delivered by four consecutive daily intraperitoneal injections. TdTomato was robustly expressed in the RPE of mice of both sexes for inductions beginning at P14 (males 90.7 ± 4.5%, females 84.7 ± 3.2%) and at 7 weeks (males 84.3 ± 7.0%, females 82 ± 3.6%). <0.6% of Muller glia also expressed tdTomato, but no tdTomato fluorescence was observed in other ocular cells or in multiple non-ocular tissues, with the exception of sparse foci in the testis. No evidence of retinal toxicity was observed in mice homozygous for the transgene induced beginning at P14 and assessed at 7-10 months. RPE-selective ablation of Tfam beginning at P14 led to reduced retinal thickness at 8 months of age and diminished retinal electrical responses at 12 months, as expected. These findings demonstrate that we have generated a mouse line with consistently efficient, tamoxifen-mediated postnatal induction of Cre recombination in the RPE and a small fraction of Muller glia. This line should be useful for temporally regulated modulation of gene function in the murine RPE.<br /> (Copyright © 2020 The Authors. Published by Elsevier Ltd.. All rights reserved.)
- Subjects :
- Animals
Disease Models, Animal
Electroretinography
Integrases biosynthesis
Macular Degeneration metabolism
Macular Degeneration pathology
Mice
Mice, Inbred C57BL
Mice, Transgenic
RNA genetics
RNA metabolism
Retinal Pigment Epithelium pathology
Tomography, Optical Coherence
Gene Expression Regulation
Integrases genetics
Macular Degeneration genetics
Retinal Pigment Epithelium metabolism
Subjects
Details
- Language :
- English
- ISSN :
- 1096-0007
- Volume :
- 202
- Database :
- MEDLINE
- Journal :
- Experimental eye research
- Publication Type :
- Academic Journal
- Accession number :
- 33264655
- Full Text :
- https://doi.org/10.1016/j.exer.2020.108370