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.
