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2014, the Core will be working (experimenting) with several labs to establish
gene targeting directly from zygote injections using the generic homing
endonuclease technology. This methodology has numerous powerful advantages over
gene targeting in ES cells. The cost and time to desired mouse line ultimately
will be dramatically reduced. We feel that this avenue of development is a very
key technology for the Core. In addition to those benefits mentioned above,
gene targeting in other animal species should become available with these
techniques. Finally, these new methodologies can be used to insert single copy
transgenes into specific genomic locations eliminating founder evaluations
required with conventional transgenesis and will allow the comparison of
transgenes with subtle differences since the site and copy number can be held
The facility's essential purpose is the provision of centralized
instrumentation and expertise for the generation, the breeding and the analysis
of both transgenic and gene-targeted animals in support of Iowa and Iowa State primary investigators. The facility
functions by providing supplies, diverse and well maintained equipment, and animal services for genetically modified
rodent production, molecular biology services, animal husbandry and project
consultant expertise. The Transgenic and Genome Manipulation (TGM) Facility is
a fee-for-service core facility of the Roy J. and Lucille A. Carver
College of Medicine. Our services are also available to outside
institutions. The TGM Core is composed of 4 personnel trained in a
variety of mouse embryo manipulations. The core conducts several methods aimed at producing and
genotyping genome-manipulated mice. The Core has three wet labs, a microscopy
suite, a tissue culture room and a molecular biology lab. Additionally, the TGM Core works in
partnership with the Multi-Tenent Facility's SPF barrier animal facility under the direction of Jim
Hynes. This facility ensures that the animal models produced by the Core are
produced in the highest quality conditions absent of pathogen load.User laboratories should be prepared to receive and maintain their
animal models once they are genotyped by the facility. At weaning all animals are transferred to the investigator's account and become the investigator's responsibility. All investigators must
have prior University Animal Care and Use Committee (UACUC) and
Recombinant DNA approval for the experiments proposed and these approvals
must be on file with the Animal Care Unit and the facility director
The TGM Core is
charged with providing a wide variety of animal services on a fee for service basis.Historically, investigators, or staff whom they support, have prepared
transgene constructs and submitted those constructs to the TGM Core for pronuclear micro injection.The
facility does the pronuclear micro injections, prepares the tail DNA
samples and screens the DNA for founder animals as a single service. Highly
experienced facility staff perform all manipulations required to
generate transgenic animals for the investigator and will identify
founder transgenic animals by PCR amplification using primers supplied
by the user. The facility will delivers a minimum of three independent
PCR-positive, transgenic founders per construct. Currently, the TGM Core offers conventional pronuclear injection to produce
transgenic animals.We have embarked on
improving this service with the acquisition of a piezo drill. Among other uses,
a piezo drill can be utilized to inject transgene-treated sperm into
unfertilized eggs and dramatically increase the rate of founder production. The
core currently has a 100% founder success rate with nearly 300 different
constructs. More information can be found by following the pronuclear injection
link on this page.
Core performs sperm cryo preservation as a vehicle to archive valuable mouse
lines and protect these lines from unexpected natural disasters such as a disease
outbreak or the loss of animal support services due to a widespread power disruption.
Sperm cryo preservation is also an alternative to retaining maintenance colonies
of valuable mice for unfunded projects. The Core’s sperm cryopreservation
service utilizes kits from The Jackson Labs. More information and a special offer can be found by
following the service description links on this page.
The TGM Core now offers
embryo rederivation to clean up both imported and in house mouse lines with
unacceptable pathogen loads. This services is offered through coordination with
the Office of Animal Resources veterinary staff. They provide some
procedural oversight when particularly dangerous mouse pathogens are involved.
To initiate this service follow the link on this page.
Core offers reanimation of cryo preserved 2-cell embryos provided that sufficient
numbers of frozen embryos are available. The core is also able to perform the
cryopreservation of 2-cell embryos. This is particularly desirable when a
mouse line can be maintained in a homozygous genotype since homozygous mice will be reanimated. At best sperm cryopreservation produces heterozygous mice. A service description link is on the the services menu for this page.
The TGM Core
provides transgene and gene targeting design and construction services. These
services are highly individualized and are conducted in very close consultation
with each investigator to insure that all possible scientific considerations
are designed into the genetic modification.Contact Phil Sanford, PhD. for more information
The TGM Core provides
ES cell expansion from cryo preserved vials and blastocyst injections to
produce germ line chimeras from gene-targeted ES cells. The Core accepts gene
targeted ES cell clones from a variety of worldwide targeted ES cell clone
repositories. Be advised that not all mouse ES cells are created equally. There
are a variety of factors that can adversely affect the quality of chimeras
produced from theses repositories. Feel free to contact Dr. Sanford for more
All services have a project status page which is updated weekly. The link to this page is found on the right hand side of the appropriate service description page.
The Core provides
extensive expert transgenic and gene targeted mouse consultation before,
during, and after project completion. Similarly, the Core designs and constructs
both transgenic and gene-targeting vectors designed by the Core director.
Please feel free to contact
the Core director regarding questions about project capability, project time lines, service fees, and billing questions.
Transgenic and Genome Manipulation Coreoffice: (319) 335-4495lab: (319) 335-4060
We can now provide microinjection services for investigators outside the State of Iowa. Contact the facility director with specific requests.
Transgenic mice were generated at the University of Iowa Transgenic and Genome Manipulation Facility directed by L. Phillip Sanford, PhD and supported in part by grants from the NIH and from the Roy J. and Lucille A. Carver College of Medicine. We wish to thank Norma Sinclair, Patricia Yarolem and Joanne Schwarting for their technical expertise in generating transgenic mice.
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