Mouse Embryonic Stem Cell and Transgenesis
Core Laboratories
Medical College of Georgia
Institute of Molecular Medicine and Genetics
The Mouse ES Cell and Transgenesis Core Laboratories provide a
centralized service of transgenic mouse production to the MCG community. It
offers services such as DNA injection into fertilized oocytes, ES cell
injection into blastocysts, strain rederivation, and embryo and sperm
cryopreservation, along with the care and housing of the mice up to the
stage where they would be transferred to the participating investigator. The
core is fully equipped to carry out these procedures. It is
located behind a specific pathogen free environment where admittance is
restricted and all food, water, cages, and bedding are autoclaved before
use.
Introduction
The transgenic mouse system:
Integration of genes into the mouse genome can serve as an experimental
system to study normal as well as altered gene expression. Such studies can
also serve to identify genes essential for normal development and generate
animal models for human diseases. Direct microinjection of DNA into
the male pronucleus of a mouse zygote has been the method most extensively
used in the production of transgenic mice. If the foreign DNA integrates
into the mouse chromosomal DNA at the one-cell stage, the transgenic animal
will contain the injected DNA in every cell, including those of the germ
line. If, however, integration occurs later, the resulting animals
will be mosaic for the presence of the injected DNA. The number of copies of
integrated DNA can vary from one to several hundred, arranged primarily in
tandem head to tail arrays.
The production of chimeric mice by introducing
Embryonic Stem (ES) cells into early embryos is an alternative approach to
generating transgenic mice. The technique utilizes the tissue culture system
to modify and select ES cells that had received an exogenous DNA of
interest. Once derived and characterized, ES cell clones may be transferred
into 3.5-day-old mouse embryos where they can differentiate into any adult
tissue type.
The Facility
The purpose of the Transgenic & ES Cell Core Laboratories is to provide a
centralized service to efficiently produce transgenic mice for basic
research. This should result in reduction in effort and cost to
participating investigators.
The core is located on the first floor of the R
& E Building. This core consists of two injections room, several animal
rooms and animal breeding rooms. The injection rooms are fully equipped to
carry out the entire procedure of making transgenic mice. The animal rooms
provide housing and breeding space for the mice involved in the transgenic
projects. The core uses sterile food and water as well as autoclaved
cages and bedding; all cages are of the microisolator type to limit the
spread of colony infection.
The entire facility is located behind a specific
pathogen free barrier where admittance is strictly limited and all personnel
must wear sterile coveralls, gloves, hats, masks, and boots.
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