GENETIC CONTROL OF MESODERM AND NEURAL CREST FORMATION

描述（改编自研究者摘要）： 该提议是为了了解分子和遗传机制， 调节细胞迁移和上皮-间充质转化， 着床后小鼠胚胎。 在本提案中，这些问题将是 通过构建和分析基因的靶向无效突变来研究 sna和slug。 这些基因是果蝇基因蜗牛的小鼠同源物， 它编码一种锌指蛋白， 果蝇胚胎发生过程中胚层的形成。 蜗牛分析 在其他脊椎动物中的同源物表明蜗牛家族基因是重要的， 在调节细胞之间的粘附相互作用，例如 参与上皮-间充质转化， 中胚层和神经嵴的产生。 的具体目标 该提议是：1）测试sna和蛞蝓基因是 正常胚胎发育所必需的，而胚胎纯合子 这些基因的无效突变将显示分层缺陷， 组织如中胚层、壁内胚层和神经嵴的迁移， 通过产生和表征在这些基因中含有无效突变的小鼠， 基因. 2)评估sna和slug突变表型的细胞自主性， 构建SNA和Slug突变体纯合的胚胎干细胞系 等位基因，并分析这些细胞在嵌合胚胎中的行为。 第三章 确定小鼠的sna和slug基因是否在功能上部分 通过构建和分析sna/slug双突变体， 四、 测定重组SNA和SLUG的DNA结合位点特异性 蛋白质，并产生针对这些蛋白质的抗体。 5)克隆 蜗牛基因家族的其他成员，并表征其 在野生型胚胎以及SNA和蛞蝓突变体胚胎中表达。

DESCRIPTION (adapted from investigator's abstract): The long term goal of this proposal is to understand the molecular and genetic mechanisms that regulate cell migration and epithelial-mesenchymal transitions in early postimplantation mouse embryos. In this proposal, these questions will be studied by constructing and analyzing targeted null mutations of the genes sna and slug. These genes are mouse homologs of the Drosophila gene snail, which encodes a zinc finger protein that is required zygotically for mesoderm formation during Drosophila embryogenesis. Analysis of snail homologs in other vertebrates suggests that snail family genes are important in regulating adhesive interactions between cells, such as the interactions involved in the epithelial-mesenchymal transition required for the generation of both the mesoderm and the neural crest. The specific aims of this proposal are: 1) Test the hypothesis that the sna and slug genes are essential for normal embryonic development, and that embryos homozygous for null mutations of these genes will show defects in delamination and migration of tissues such as mesoderm, parietal endoderm and neural crest, by generating and characterizing mice containing null mutations in these genes. 2) Assess the cell autonomy of sna and slug mutant phenotypes by constructing embryonic stem cell lines homozygous for sna and slug mutant alleles, and analyzing the behavior of these cells in chimeric embryos. 3) Determine whether the sna and slug genes in mice are in part functionally redundant by constructing and analyzing sna/slug double mutants. 4) Determine the DNA binding site specificity of recombinant SNA and SLUG proteins, and generate antibodies specific for these proteins. 5) Clone additional members of the snail gene family, and characterize their expression in both wild type embryos and in sna and slug mutant embryos.