A Microinjection Apparatus and Associated Equipment for Creation of Transgenic Mice

用于创建转基因小鼠的显微注射装置和相关设备

• 批准号: 9602951
• 负责人: Kevin Jones
• 金额: $ 7.32万
• 依托单位: University of Colorado at Boulder
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 1997
• 资助国家: 美国
• 起止时间: 1997-04-01 至 1999-03-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=9602951&HistoricalAwards=false
• 关键词: Microinjection; Apparatus; Associated; Equipment; Creation

Funds are requested for the purchase of an embryo microinjection apparatus and associated equipment to be used in the production of transgenic mice. This apparatus is to be used primarily for production of transgenic mice using two technologies: pronuclear injection of DNA and blastocyst/morula injection of embryonic stem cells. These are now well-established methods with which all of the primary users are quite familiar. The apparatus will also be used for transplantation experiments (e.g. mitochondrial transplantation). This apparatus is to be housed in a new transgenic facility, the construction of which is to be completed by July 1, 1996. This facility will serve faculty members of the MCD Biology and other departments on the University of Colorado Boulder campus including Biochemistry and the Institute for Behavioral Genetics. As a reflection of the diverse research interests of these faculty in multiple areas of biology, the animals to be created will address a wide range of scientific issues. Areas under investigation include the functions of neurotrophic factors in the development and maintenance of the nervous system, the function of Wnt-family genes in the morphogenesis of the limb and nervous system, the molecular basis of muscle formation and contractility, the function of homologues of the Drosophila mago nashi gene in mouse germ-cell development, and the localization of quantitative trait loci. This apparatus will be a core piece of equipment and is essential in the production of transgenic mice. The analysis of transgenic mice makes feasible the investigation of gene function in a mammal through creation of animals having a defined genetic perturbation. Creation of transgenic mice by pronuclear injection is the only widely used method of integration of cloned DNA into random sites in the mouse genome. Production of transgenic mice using embryonic stem cells modified by gene targeting through blastocyst injection is presently favored (ove r morula aggregation) by most practitioners. Thus, a microinjection apparatus is an essential feature of any facility for the production of transgenic mice.

要求拨款购买胚胎显微注射装置和用于生产转基因小鼠的相关设备。该装置主要用于通过两种技术生产转基因小鼠：DNA原核注射和胚胎干细胞囊胚/桑葚胚注射。现在这些都是行之有效的方法，所有主要用户都非常熟悉。该仪器也将用于移植实验（如线粒体移植）。这个装置将被安置在一个新的转基因设施中，该设施的建设将于1996年7月1日完成。该设施将为MCD生物学和科罗拉多大学博尔德校区其他部门的教职员工提供服务，包括生物化学和行为遗传学研究所。作为这些教师在生物学多个领域的不同研究兴趣的反映，将要创建的动物将解决广泛的科学问题。研究领域包括神经营养因子在神经系统发育和维持中的功能、wnt家族基因在肢体和神经系统形态发生中的功能、肌肉形成和收缩的分子基础、果蝇马格纳希基因同源物在小鼠生殖细胞发育中的功能以及数量性状位点的定位。该装置将是一个核心设备，是生产转基因小鼠必不可少的。转基因小鼠的分析使得通过创造具有确定的遗传扰动的动物来研究哺乳动物的基因功能成为可能。通过原核注射建立转基因小鼠是唯一广泛使用的将克隆DNA整合到小鼠基因组随机位点的方法。利用胚泡注射基因靶向修饰的胚胎干细胞生产转基因小鼠目前受到大多数从业者的青睐（而不是桑葚胚聚集）。因此，显微注射装置是生产转基因小鼠的任何设施的基本特征。