Walter Guy Wiles1, Zhongming Mou2, Yang Du3, Alyssa B Long4, Christopher D Scharer5, Birdal Bilir2, Demetri D Spyropoulos6, Nancy A Jenkins7, Neal G Copeland7, W David Martin8, Carlos S Moreno9. 1. Winship Cancer Institute, Emory University School of Medicine, Atlanta, GA, U.S.A. 2. Department of Pathology and Laboratory Medicine, Emory University School of Medicine, Atlanta, GA, U.S.A. 3. Department of Pediatrics, Uniformed Services University of the Health Sciences, Bethesda, MD, U.S.A. 4. Department of Human Genetics, Emory University School of Medicine, Atlanta, GA, U.S.A. 5. Department of Microbiology and Immunology, Emory University School of Medicine, Atlanta, GA, U.S.A. 6. Department of Pathology and Laboratory Medicine, Medical University of South Carolina, Charleston, SC, U.S.A. 7. Cancer Research Program, The Methodist Hospital Research Institute, Houston, TX, U.S.A. 8. Winship Cancer Institute, Emory University School of Medicine, Atlanta, GA, U.S.A. Department of Pathology and Laboratory Medicine, Emory University School of Medicine, Atlanta, GA, U.S.A. 9. Winship Cancer Institute, Emory University School of Medicine, Atlanta, GA, U.S.A. Department of Pathology and Laboratory Medicine, Emory University School of Medicine, Atlanta, GA, U.S.A. cmoreno@emory.edu.
Abstract
BACKGROUND: Sox4 is an essential gene, and genetic deletion results in embryonic lethality. In an effort to develop mice with tissue-specific deletion, we bred conditional knockout mice bearing LoxP recombination sites flanking the Sox4 gene, with the LoxP sites located in the Sox4 5'UTR and 3'UTR. RESULTS: The number of mice homozygous for this LoxP-flanked conditional knockout allele was far below the expected number, suggesting embryonic lethality with reduced penetrance. From over 200 animals bred, only 11% were homozygous Sox4(flox/flox) mice, compared to the expected Mendelian ratio of 25% (p<0.001). Moreover, there was a significant reduction in the number of female Sox4(flox/flox) mice (26%) relative to male Sox4(flox/flox) mice (p=0.0371). Reduced Sox4 expression in homozygous embryos was confirmed by in-situ hybridization and Quantitative real-time polymerase chain reaction (QPCR). CONCLUSION: LoxP sites in the 5' and 3' UTR of both alleles of Sox4 resulted in reduced, but variable expression of Sox4 message.
BACKGROUND:Sox4 is an essential gene, and genetic deletion results in embryonic lethality. In an effort to develop mice with tissue-specific deletion, we bred conditional knockout mice bearing LoxP recombination sites flanking the Sox4 gene, with the LoxP sites located in the Sox4 5'UTR and 3'UTR. RESULTS: The number of mice homozygous for this LoxP-flanked conditional knockout allele was far below the expected number, suggesting embryonic lethality with reduced penetrance. From over 200 animals bred, only 11% were homozygous Sox4(flox/flox) mice, compared to the expected Mendelian ratio of 25% (p<0.001). Moreover, there was a significant reduction in the number of female Sox4(flox/flox) mice (26%) relative to male Sox4(flox/flox) mice (p=0.0371). Reduced Sox4 expression in homozygous embryos was confirmed by in-situ hybridization and Quantitative real-time polymerase chain reaction (QPCR). CONCLUSION:LoxP sites in the 5' and 3' UTR of both alleles of Sox4 resulted in reduced, but variable expression of Sox4 message.
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