SKI PROTO-ONCOGENE AND MOUSE DEVELOPMENT

Ski 原癌基因和小鼠发育

• 批准号: 2889093
• 负责人: CLEMENCIA COLMENARES
• 金额: $ 10.39万
• 依托单位: CLEVELAND CLINIC FOUNDATION
• 依托单位国家: 美国
• 财政年份: 1996
• 资助国家: 美国
• 起止时间: 1996-08-01 至 2001-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2889093
• 关键词: DNA binding protein; RNase protection assay; cell adhesion; cell differentiation; confocal scanning microscopy; cytogenetics; developmental genetics; developmental neurobiology; embryo /fetus cell /tissue; embryonic stem cell; gene expression; gene mutation; gene targeting; genetically modified animals; immunocytochemistry; laboratory mouse; mammalian embryology; neural plate /tube; neurogenetics; northern blottings; polymerase chain reaction; protooncogene; scanning electron microscopy; southern blotting; transcription factor; western blottings

Genes that encode regulators of gene expression have proved to play a central role in directing mammalian growth and development. The ski proto-oncogene encodes a nuclear protein shown to bind DNA and regulate transcription. Its biological activities impinge on both growth and differentiation pathways in many cell lineages, including the myogenic and hematopoietic systems. To study the physiological functions of ski in vivo, we have disrupted the gene in murine embryonic stem cells, and introduced this mutation into the mouse germ line. Our data shows that ski is essential for normal development and survival, since mice homozygous for the ski mutation die at birth. In addition to generalized growth retardation, the major defect is in the central nervous system. Affected mice are born with exencephaly, a condition that normally results from failure of neural tube closure during neurulation. This result is consistent with studies showing high mRNA expression in the developing neural tube and neural crest. We propose to study the role of ski in neural development by studying the morphogenetic events and molecular mechanisms that are defective in its absence. Our specific aims include: developmental and histological studies of the defective phenotype as well as analyses of expression of other genes such as domain-specific transcription factors and cell adhesion molecules; generation of chimeras using homozygous mutant ES cell lines to study the specific cell types and mechanisms that require ski function; and analysis of homozygous mice expressing a partially defective mutant ski gene. Neural tube defects (NTDs) are some of the most common human congenital abnormalities, and their etiology is almost certainly multifactorial. It has been estimated that 10 or 12 genes control the susceptibility to NTDs, and our studies suggest that ski may be one of this group of genes. The proposed work will test this hypothesis, and should contribute to the understanding the numerous genetic and molecular interactions required for normal neural tube development in mammals.

编码基因表达调节因子的基因已被证明发挥了 指导哺乳动物生长和发育中的核心作用。滑雪 原始癌基因编码显示DNA并调节的核蛋白 转录。它的生物学活动影响了增长和 许多细胞谱系中的分化途径，包括肌原性 和造血系统。研究滑雪的生理功能 在体内，我们破坏了鼠类胚胎干细胞中的基因， 将此突变引入了小鼠种系。我们的数据表明 滑雪对于正常发育和生存至关重要，因为小鼠 滑雪突变出生时纯合子死亡。除了广义 生长迟钝，主要缺陷在中枢神经系统中。 受影响的小鼠出生于excephaly，这种情况通常是 神经管闭合期间的失败导致。这 结果与显示出高mRNA表达的研究一致 开发神经管和神经波峰。 我们建议通过研究滑雪在神经发育中的作用 形态发生事件和分子机制有缺陷 缺席。我们的具体目的包括：发展和组织学 对缺陷表型的研究以及表达的分析 其他基因，例如域特异性转录因子和细胞 粘附分子；使用纯合突变体产生嵌合体 细胞系研究需要的特定细胞类型和机制 滑雪功能；和分析部分表达部分的纯合小鼠 有缺陷的突变滑雪基因。神经管缺陷（NTD）是其中的一些 最常见的人类先天性异常及其病因几乎是 当然是多因素。据估计，10或12个基因 控制对NTD的敏感性，我们的研究表明滑雪 成为这一基因之一。拟议的工作将测试 假设，应该有助于理解众多 正常神经管所需的遗传和分子相互作用 哺乳动物的发展。