Basic Biology of Aging at the University of Washington

Transgenic Animal Model Development Core

Description

The Transgenic Animal Model Development Core is designed to support investigators doing biology of aging research by creating mice that have been genetically altered by inserting a new gene, removing a normal gene, or introducing specific base pair mutations. The use of genetically engineered mouse models has become one of the most exciting approaches to discovering the functions and interactions of genes in mammals. At the University of Washington Nathan Shock Center, this transgenic technology is used to develop new animal models for studying genetic mechanisms of the aging process.

During the past several years, transgenic mouse production has focused on constructs with enhanced defense against free radical injury in aging (e.g., catalase, superoxide dismutase, glutathione S-transferase), Werner Syndrome, adult onset diabetes, Alzheimer's Disease, thrombospondin, and rheumatoid arthritis in aging. In addition, the core has concentrated an appreciable portion of effort into gene-targeting methodologies for the production of gene knockouts, knock-ins, and targeted ES transgenics. This included work to generate mouse models of Werner's Syndrome, models for study of presenillin genes related to Alzheimer's Disease and study of models of thrombospondin in aging.  

The Transgenic Animal Model Development Core is administered through the Transgenic Resources Program (TRP), a cost center within the Department of Compartment of Comparative Medicine, dedicated to producing and maintaining genetically engineered mice. The success in producing genetically engineered lines is the result of work to provide a state-of-the-art transgenic technology resource which is well-equipped and well-staffed to develop the next generation of mouse models for numerous research projects. Over the last 3 years, the TRP has created about 300 new lines of mice by microinjection, cryopreserved 56 lines of mice, rederived (by embryo rederivation and/or IVF) 24 lines of mice, and performed 8 ovary transfer surgeries. We have established new services for sperm cryopreservation as well as mouse rederivation via in vitro fertilization. This has been a crucial step, allowing investigators to obtain transgenic mouse lines from publicly- funded consortiums such as the KOMP, EUCOMM, and NORCOMM which produce and distribute genetically engineered mice at a low subsidized cost.

Inquiries regarding these services are welcome. A very cost effective cryopreservation procedure is available to preserve important mouse lines. If you are considering making a new transgenic or gene targeted mouse line, please contact us and we will find out if it already exists and if it is available. We can do embryo or ovary transfers or sperm in vitro fertilization to rederive the line. If it does not exist, we will make it for you. 

Background

The isolation and manipulation of mammalian genes are of utmost importance to the biology and medicine of aging because of the contributions these studies can make to the understanding of physiology and development. Techniques for introducing foreign genes and/or mutations into the mouse germ line provide novel approaches for modeling human genetic and chronic degenerative diseases.

Since the initial report in 1980 describing transgenic mice, methods for the direct microinjection of DNA into the pronuclei of fertilized embryos have become established, allowing foreign genes to be incorporated into somatic germline tissues, with expression of these elements in the progeny of founder mice. The creation of "transgenic" animals that make a specified gene product presents a spectrum of opportunities for basic studies in molecular pathogenesis and pre-clinical investigations applicable to a wide variety of medical problems of aging.

An additional gene transfer technology initially developed in the 1980's involved the use of mouse embryonic stem (ES) cells isolated from the early embryo. The capacity of ES cells to undergo differentiation makes them useful for investigating the effects of genetic modifications of either the gain of function or loss of function. These pluripotent genetically modified ES cells can then be used to make mice with deleted genes (gene knockout) or targeted mutagenesis of genes thought to be involved in the aging process. The University of Washington Aging Program provides a focused level of expertise and resources to enhance and facilitate the development of transgenic animal models using ES cell transfer technology.

Facilities

The Transgenic Resources Program is located within the K-wing barrier facility of the University of Washington Health Sciences Center. This core resource facility has a dedicated allocation of laboratory and rodent housing space within an area that was custom designed by the Department of Comparative Medicine for a transgenic mouse facility. The space consists of 3 dedicated procedure rooms used for embryo microinjection, embryo and sperm cryopreservation, as well as mouse rederivation and in vitro fertilization. The animals receiving these genetic engineering procedures are housed across the hall from the laboratories, in specific pathogen free (SPF) rooms designed to accommodate rodent cages individually ventilated with filtered air, and cage changing performed in laminar flow hoods. In addition, the core also has a tissue culture room located in the T-wing for use in gene targeting experiments using mouse ES cells. 

Personnel

Warren Ladiges, Director, is a DVM with extensive experience in use of murine models in biomedical research

Robert Hunter, Program Manager, Transgenic Resources Program

Serina Tsang, Research Scientist, ES cell gene targeting

Shoshana Maslan, Research Scientist, microinjection, cryopreservation, and mouse rederivation

La’Akea Siverts, Research Scientist, microinjection, cryopreservation, and mouse rederivation