ROLES OF SOX C GENES IN SKELETOGENESIS

SOX C 基因在骨骼形成中的作用

• 批准号: 7277746
• 负责人: VERONIQUE M LEFEBVRE
• 金额: $ 33万
• 依托单位: CLEVELAND CLINIC LERNER COM-CWRU
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-09-01 至 2011-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7277746
• 关键词: Affect; Alleles; Antibodies; Biological Assay; Birth; Cell Differentiation process; Cell Lineage; Cells; Chondroblast; Chondrocytes; Cleft Palate; Complement; Congenital Heart Defects; Cultured Cells; DNA Binding; Data; Defect; Digit structure; Disease; Embryo; Fetus; Gene Expression; Gene Targeting; Genes; Genetic; Genetic Transcription; HMG Domain; HMG-Box; Immunoprecipitation; In Vitro; Jaw; Laboratories; Mesenchymal; Metatarsal bone structure; Molecular; Mus; Mutant Strains Mice; Osteoblasts; Pattern; Primary Cell Cultures; Property; Proteins; Regulator Genes; Research; Research Personnel; Role; Screening procedure; Site; Skeletal system; Skeleton; Staging; Standards of Weights and Measures; System; Testing; Trans-Activators; Transactivation; Transgenes; Transgenic Mice; Western Blotting; Work; cell motility; citrate carrier; in vitro Assay; in vivo; insight; lumbar vertebra bone structure; precursor cell; programs; research study; transcription factor; vertebra body

DESCRIPTION (provided by applicant): This project will uncover the roles of Sox4, Sox11, and Sox12 in skeletogenic cells in vivo. These three highly related proteins form the subfamily C of Sry-related HMG box (Sox) transcription factors. Their genes are co-expressed in skeletogenic mesenchymal cells throughout the mouse embryo, and remain differentially expressed when these cells undergo chondrocyte and osteoblast differentiation. While the roles of Sox12 in vivo remain unknown, mice lacking Sox4 and/or Sox11 have started to reveal important roles for these two genes in multiple steps of skeletogenesis. The molecular roles of Sox C proteins remain largely unknown, but preliminary studies have suggested that they may bind DNA and activate transcription similarly but with different efficiencies. These data raise the hypothesis that Sox C proteins may have redundant, complementary, or distinct molecular roles in controlling skeletogenic cell fate and differentiation. To test this hypothesis, Aim 1 is to further study the roles of the Sox C proteins in mouse embryo skeletogenesis. Mice carrying Sox11 and Sox12 conditional null alleles will be generated using the same strategy that was used to generate a Sox4 conditional null allele. Well-established Cre transgenes will be used to specifically inactivate the genes in the entire embryo or at specific steps during skeletogenesis. Skeletal defects will be characterized at the morphological, cellular and molecular levels, and primary cell culture experiments will be carried out to complement in vivo studies. Aim 2 is to characterize the molecular roles of Sox C proteins in skeletogenic cells. The DNA-binding and transactivation properties of the three Sox C proteins will be further studied using standard in vitro and cell culture assays. Potential target genes will be identified by gene expression array screening. The action of Sox C proteins on target gene regulatory sequences will be studied using DNA-binding and transactivation assays in vitro and in vivo. It is anticipated that this study will identify Sox C proteins as essential determinants of skeletogenic cells from pluripotent precursor stages until late maturation in specific cell lineages, and will thereby provide significant insights into molecular mechanisms involved in normal skeletogenesis and in genetic and acquired diseases of the skeleton.

描述（由申请人提供）：该项目将揭示Sox4，Sox11和Sox12在体内骨骼生成细胞中的作用。这三种高度相关的蛋白质形成了与SRY相关的HMG盒（SOX）转录因子的亚家族C。它们的基因在整个小鼠胚胎中共表达在骨骼发育性间充质细胞中，并在这些细胞发生软骨细胞和成骨细胞分化时保持差异表达。尽管Sox12在体内的作用仍然未知，但缺乏Sox4和/或Sox11的小鼠已经开始在骨骼生成的多个步骤中揭示这两个基因的重要作用。 Sox C蛋白的分子作用在很大程度上尚不清楚，但是初步研究表明它们可以结合DNA并类似地激活转录，但具有不同的效率。这些数据提出了这样的假设：Sox C蛋白可能在控制骨骼发明细胞命运和分化中具有冗余，互补或不同的分子作用。为了检验该假设，目标1是进一步研究Sox C蛋白在小鼠胚胎骨骼生成中的作用。携带Sox11和Sox12条件无效等位基因的小鼠将使用与生成Sox4条件无效等位基因相同的策略生成。建立的CRE转基因将用于特异性使整个胚胎中的基因或骨骼生成过程中的特定步骤中的基因。骨骼缺陷将在形态，细胞和分子水平上进行表征，将进行原代细胞培养实验以补充体内研究。目的2是表征Sox C蛋白在骨架细胞中的分子作用。三种SOX C蛋白的DNA结合和反式激活特性将使用标准的体外和细胞培养测定进一步研究。潜在靶基因将通过基因表达阵列筛选来鉴定。 SOX C蛋白对靶基因调节序列的作用将在体外和体内使用DNA结合和反式激活测定。可以预料，这项研究将识别出Sox C蛋白是从多能前体阶段进行骨骼生成细胞的重要决定因素，直到在特定细胞谱系的晚期成熟，从而为涉及正常骨骼生成以及骨架的遗传疾病中涉及的分子机制提供了重要的见解。