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E.J. Talamas, D.J. Sidjanin, L. Jackson, M. Koeberl, T. Jackson, J.L. McElwee, M.M. Jablonski, B. Chang; Early Transposable Element Insertion in Intron 9 of Hsf4 Gene Results in Autosomal Recessive Cataracts in Lop11 and Ldis1 Mice . Invest. Ophthalmol. Vis. Sci. 2006;47(13):4599.
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Lens opacity (lop11) is an autosomal recessive mouse cataract locus that has been mapped to mouse chromosome 8. Evaluation of the mouse genome map of the lop11 critical region identified Heat Shock Transcription Factor 4. The goal of this study is to evaluate Hsf4 as a candidate gene for lop11.
mRNA and protein were isolated from P21 lop11 and C57BL/6 mouse lenses. RT–PCR, northern blotting and western blots were done using standard methods. For sequence analysis genomic DNA from lop11 and ldis1 mice was isolated from spleens. Primer design, PCR and sequencing were all performed using standard methods.
RT–PCR from lop11/lop11 mRNA with primers specific to Hsf4 exons 1 through 9 generated cDNA products whereas primers specific for Hsf4 exons 10 through13 did not yield any cDNA products in lop11/lop11. Norhtern blot analysis was consistent with RT–PCR data. Sequencing of the genomic Hsf4 region identified an early transposable element (ETn) in intron 9 inserted 61 bp upstream from exon 10. The same mutation was also identified in ldis1, another mouse cataract locus on chromosome 8. Western blot analysis identified a 40 kd Hsf4 immunoreactive band in lop11/lop11 and wild type 55kDa Hsf4 was absent.
Molecular evaluation of Hsf4 revealed an ETn in intron 9 inserted 61 bp upstream of the intron/junction in both lop11 and ldis1 mice. In lop11 ETn insertion altered splicing and expression of the Hsf4 gene, resulting in the truncated Hsf4 protein. The lop11 mouse is an excellent resource for evaluating the role of Hsf4 for transparency of the lens.
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