LAC OPERON CONTROL OF HGH EXPRESSION IN GENE THERAPY

基因治疗中 LAC 操纵子对 HGH 表达的控制

• 批准号: 2134184
• 负责人: ROBERTO E IZQUIERDO
• 金额: $ 7.56万
• 依托单位: ALBANY MEDICAL COLLEGE
• 依托单位国家: 美国
• 财政年份: 1994
• 资助国家: 美国
• 起止时间: 1994-08-01 至 1999-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2134184
• 关键词: galactosides; gene expression; gene induction /repression; laboratory mouse; lac operon; somatotropin; tissue /cell culture; transfection

DESCRIPTION: The training plan for the candidate will focus on two phases. Phase I will involve approximately five or six formal courses taking about 45 percent of the candidate's time during the first two years of the program. In addition, during this same time period, the candidate will be spending approximately 45 percent of his time on research-related activities and about 10 percent on clinical responsibilities. His progress will be monitored by the sponsor, as well as a faculty advisory committee which will serve as the Advisory Committee for the annual review, and the evaluation of the candidate's formal research plan which will be developed during the first two years of his training. The transition from phase I to phase II will be controlled by this Faculty Advisory Committee. The research outlined in this application centers on the idea that there has been no attempt to stringently regulate the expression of a transferred gene in an animal. The candidate believes that the enzymes important for lactose metabolism (lac) or lac operon system, a gene repressor/operator system of Escherichia coli (E. coli) could be of value in this regard. It has been established that the lac repressor and operator are functional in mammalian cells in tissue culture, including murine C2C12 myoblasts, and that the inducing agent, isopropyl beta-D- thiogalactoside (IPTG), can reverse repression.Further, the lac system can achieve stringent repression of an operator-linked gene that is stably integrated into a mammalian chromosome, and at least several hundred-fold induction of expression can be achieved with IPTG. Induction is fully reversible with removal of IPTG, and IPTG has been shown to be nontoxic in mice and to be biologically active when administered intraperitoneally. The project proposes to introduce the E. coli lac regulatory system linked to the human growth hormone (hGH) gene into murine C2C12 myoblasts, and implant these engineered cells into mice. The hypothesis is that systemic expression of hGH could then be regulated by intraperitoneal administration of varying amounts of the inducing agent, IPTG. Future work will be to apply gene transfer treatment to humans with inherited protein hormone deficiency states such as idiopathic hGH deficiency.

产品说明： 候选人的培训计划将侧重于两个阶段。 一期 将涉及大约五个或六个正式课程， 候选人在前两年的时间百分比 程序. 此外，在同一时间段内，候选人将 他将大约45%的时间花在与研究相关的工作上， 约10%的临床责任。 他 赞助商和教师顾问将监督进展情况 委员会将作为年度咨询委员会， 审查，并评估候选人的正式研究计划， 将在他训练的头两年里得到发展。 的 从第一阶段到第二阶段的过渡将由该学院控制 咨询委员会。 在这个应用程序中概述的研究中心的想法， 并没有试图严格地调节 转基因动物 候选人认为酶 对乳糖代谢（lac）或lac操纵子系统重要基因 大肠杆菌（Escherichia coli，E.大肠杆菌）可能是有价值的 在这方面 已经确定lac阻遏物和 操纵子在组织培养的哺乳动物细胞中起作用，包括 小鼠C2 C12成肌细胞，并且诱导剂异丙基β-D- 硫代半乳糖苷（IPTG）可以逆转阻遏。 可以实现操纵子连锁基因的严格抑制， 稳定整合到哺乳动物染色体中，并且至少几个 用IPTG可以实现百倍表达诱导。 诱导在去除IPTG的情况下是完全可逆的，并且IPTG已经被纯化。 在小鼠中显示无毒， 腹膜内给药。 该项目建议引入E.大肠杆菌乳糖调节系统 将人生长激素（hGH）基因连接到小鼠C2 C12中 成肌细胞，并将这些工程细胞植入小鼠体内。 的假设 hGH的系统性表达可以通过 腹膜内施用不同量的诱导剂， IPTG诱导 未来的工作将是将基因转移治疗应用于人类 遗传性蛋白激素缺乏症，如特发性hGH 缺陷