A Microinjection Apparatus and Associated Equipment for Creation of Transgenic Mice

请拨资金购买用于生产转基因小鼠的胚胎显微注射设备和相关设备。该设备将主要用于两种技术的转基因小鼠的生产：DNA原核注射和胚胎干细胞囊胚/桑椹注射。这些现在都是所有主要用户都非常熟悉的成熟方法。该仪器还将用于移植实验(例如线粒体移植)。这台仪器将被安置在一个新的转基因设备中，该设备的建造将于1996年7月1日完成。这个设施将为科罗拉多大学博尔德分校MCD生物学和其他系的教职员工服务，包括生物化学和行为遗传学研究所。作为这些教员在生物学多个领域的不同研究兴趣的反映，将创造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Mago nashi基因同源基因在小鼠生殖细胞发育中的功能，以及数量性状基因的定位。该设备将是核心设备，在转基因小鼠的生产中至关重要。对转基因小鼠的分析使通过创造具有明确遗传扰动的动物来研究哺乳动物的基因功能成为可能。通过原核注射创造转基因小鼠是唯一被广泛使用的将克隆的DNA整合到小鼠基因组中随机位置的方法。目前，大多数从业者倾向于使用基因打靶的胚胎干细胞通过胚泡注射生产转基因小鼠(即桑椹胚聚集)。因此，显微注射设备是任何生产转基因小鼠的设施的基本特征。

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.