TARGETED MUTATIONS IN THE ABL TYROSINE KINASE GENE

ABL 酪氨酸激酶基因的靶向突变

• 批准号: 6613894
• 负责人: STEPHEN Paine GOFF
• 金额: $ 15.94万
• 依托单位: COLUMBIA UNIVERSITY HEALTH SCIENCES
• 依托单位国家: 美国
• 财政年份: 2002
• 资助国家: 美国
• 起止时间: 2002-08-01 至 2003-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6613894
• 关键词: alleles; animal genetic material tag; biological signal transduction; bone development; cell cycle proteins; cell differentiation; developmental genetics; embryonic stem cell; gene mutation; gene targeting; genetically modified animals; laboratory mouse; mitogens; mutant; osteoblasts; osteoporosis; phenotype; protein structure function; protein tyrosine kinase; protooncogene; tissue /cell culture

This proposal describes experiments using the tools of mouse genetics to determine the in vivo functions of the c-abl proto-oncogene. The c-abl gene product is a cytoplasmic tyrosine kinase important for signal transduction and control of the cell cycle. Knock-out mice deficient in c- abl, generated in the previous funding period, exhibit a number of phenotypes: perinatal lethality, runting, bone abnormalities, and defects in early lymphoid cell lineages, notably a sensitivity to apoptotic stimuli, Several genetic studies of these mice are proposed: First, the osteoporosis causes by the c-abl deficiency will be examined; osteoblasts will be cultured from mutant and control animals, and tested for their ability to respond to a panel of growth factors. Second, new alleles of c- abl will be generated by gene targeting ("knock-in" experiments). One of these alleles will permit the conditional deletions of the gene at selected times in development and in selected tissues; others will express altered forms of c-abl lacking particular domains. Examination of these mice should help determine the functions of each pathway emanating from c- abl. Third, breeding will be used to generate compound mutant mice lacking c-abl and other genes with related functions, including the abi-1 and -2 genes, encoding Abl-interacting proteins, and ret-, a transmembrane receptor kinase under investigation by the Constantini laboratory. Finally, a germ-line mutation of a new member of the Abi family, a mammalian homologue of the yeast Cdc15 gene, will be generate by gene targeting, and the effects of the mutation alone and with other knock-out mutations will be characterize. These studies should help define the many diverse functions of c-abl in mammalian development and physiology.

该提案描述了使用小鼠遗传学工具进行的实验 确定 c-abl 原癌基因的体内功能。 c-abl 基因产物是一种对信号重要的细胞质酪氨酸激酶 细胞周期的转导和控制。基因敲除小鼠缺乏 c- abl 是在上一个资助期间产生的，展示了一些 表型：围产期致死率、矮小、骨骼异常和缺陷 在早期淋巴细胞谱系中，特别是对细胞凋亡的敏感性 刺激，建议对这些小鼠进行几项遗传学研究：首先， 将检查由 c-abl 缺乏引起的骨质疏松症；成骨细胞 将从突变动物和对照动物中培养，并测试它们的 对一组生长因子做出反应的能力。其次，c的新等位基因- abl 将通过基因靶向（“敲入”实验）产生。之一 这些等位基因将允许有条件地删除该基因 选定的发育时间和选定的组织；其他人会表达 缺乏特定结构域的 c-abl 的改变形式。检查这些 小鼠应该有助于确定来自 c-的每个通路的功能 abl。第三，育种将用于产生缺乏基因的复合突变小鼠。 c-abl和其他具有相关功能的基因，包括abi-1和-2 基因，编码 Abl 相互作用蛋白和 ret-，一种跨膜蛋白 Constantini 实验室正在研究受体激酶。 最后，Abi 家族新成员的种系突变， 酵母Cdc15基因的哺乳动物同源物，将由基因产生 靶向，以及单独突变和其他敲除突变的影响 突变将被表征。这些研究应该有助于定义许多 c-abl 在哺乳动物发育和生理学中的多种功能。