Transgenic and Genome Manipulation Facility

  • Transgenic and Genome Manipulation Facility
  • The Facility

    Microinjection  

     

    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.

    The TGM 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.

    Similarly, the 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 information.

     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.

    Beginning in 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 constant. 

     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.

     Core Director Contact Information:

    L. Philip Sanford, Ph.D.

    Director, Transgenic and Genome Manipulation Core

    l-sanford@uiowa.edu

    office: (319) 335-4495

     lab: (319) 35-4060

    cell: (319) 430-7618  

    Note:

    We can now provide microinjection services for investigators outside the State of Iowa. Contact the facility director with specific requests.

    Publication Acknowledgement:

    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.