A Microinjection Apparatus and Associated Equipment for Creation of Transgenic Mice

要求提供资金，用于购买胚胎显微注射装置和相关设备，用于生产转基因小鼠。该设备主要用于使用两种技术生产转基因小鼠：DNA的原核注射和胚胎干细胞的胚泡/桑椹胚注射。这些都是目前所有主要使用者都非常熟悉的既定方法。该装置还将用于移植实验（例如线粒体移植）。该装置将被安置在一个新的转基因设施中，该设施将于1996年7月1日完工。该设施将为MCD生物学和科罗拉多大学博尔德分校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，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mogo nashi基因同源物在小鼠生殖细胞发育中的功能，以及数量性状基因座的定位。 该装置将是设备的核心部件，在生产转基因小鼠中至关重要。转基因小鼠的分析使得通过产生具有确定的遗传扰动的动物来研究哺乳动物中的基因功能成为可能。通过原核注射产生转基因小鼠是唯一广泛使用的将克隆DNA整合到小鼠基因组中的随机位点的方法。 使用通过胚泡注射的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（超过桑椹胚聚集）。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。

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.