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H. Chen, Z. Tong, J. Kaminoh, Z. Yang, L. Luo, J. Cameron, J. Zeng, M. Mihaila, K. A. Howes, K. Zhang; Generation and Characterization of Elovl4 Y270X Knockin Mouse. Invest. Ophthalmol. Vis. Sci. 2007;48(13):4489.
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© ARVO (1962-2015); The Authors (2016-present)
ELOVL4 was first identified as a disease-causing gene in Stargardt macular dystrophy. To date, three ELOVL4 mutations have been identified including Y270X, all of which result in truncated proteins. However, the specific role of ELOVL4 in photoreceptors and the degenerative events caused by dominant ELOVL4 mutations are not well understood. The purpose of this study is to generate Y270X knockin mice which will provide a mouse model to examine the dominant pathogenic mode of degeneration in human STGD3 patients.
A mouse Elovl4 knockin targeting construct was generated containing a neo cassette flanked by loxP sites, and a Y270X mutation and HA tagged Elovl4 fragment introduced into exon 6. For gene targeting, the targeting vector was electroporated into mouse embryonic stem (ES) cells derived from 129. Clones in which homologous recombination resulted in targeted replacement of exon 6 were identified by PCR and by Southern blotting. Chimeric mice will be bred with wild type C57Bl/6 mice for germline transmission of the recombinant Elovl4 gene.
Twenty-one positive recombinant ES cell clones have been identified. Injections have yielded eight chimeric mice so far. Chimeric mice are currently being mated with C57Bl/6 mice to generate heterozygous knockin mouse.
We have generated chimeric mice which will be used to generate the Elovl4 Y270X knockin mutation. Characterization of retinal phenotypes associated with Y270X knockin mutation will be presented. These mice will provide an important animal model for examining autosomal dominant macular degeneration in STGD patients.
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