Transgenic Core Facility
The Transgenic Core consists of a laboratory for procedures involving production and maintenance of transgenic mouse strains, a tissue culture facility for all procedures involving Embryonic Stem (ES) cells and gene targeting experiments, and a transgenic barrier holding facility.
The Core is under the direction of Dr. Weiler, who has extensive experience in the production of genetically altered mice, and includes three additional staff skilled in various aspects of transgenic animal production and related procedures.
The laboratory is located in the vivarium of the Translational and Biomedical Research Center at MCW and is equipped to perform all procedures involving production and maintenance of transgenic or knock-out mice. Instrumentation includes 3 injection workstations (Nikon inverted phase contrast microscope with Hoffman modulation optics, temperature-controlled injection stage, air-suspended injection table, Eppendorf motor-driven injection system), microforge, needle puller, Nikon stereo zoom microscope, surgical microscope, CO2-incubator, laminar flow hood for surgical procedures, and a cell fusion instrument for tetraploid embryo production.
Additional workspace equipped for cryopreservation and gene targeting is located at the Versiti Blood Research Institute.
Liquid nitrogen tanks and an automated cell freezer for cryopreservation of fertilized oocytes are available as shared instrumentation through the Transgenic Core. Over the past three years, the Core has successfully performed 24 different gene targeting experiments, produced more than 14 germline-competent ES cell aggregation chimeras and gene targeted mouse strains, generated transgenic animals via zygote micro-injection, and assisted in routine animal procedures such as re-derivation of mouse strains. All services (including gene targeting, blastocysts injections, and zygote microinjections) are available to MCW-associated Investigators on a fee-for-service basis.
Protocols
A number of very useful protocols for the generation and analysis of transgenic animals are described in Manipulating the Mouse Embryo: a laboratory manual by B. Hogan et al., 2nd edition or newer, published by Cold Spring Harbor Laboratory press (ISBN 0-87969-384-3).
Forum
Quality Control Experiment To Ascertain The Germline Potential of E14Tg2a.4 ES Cells (PDF)
Director

Hartmut Weiler, PhD
Associate Professor
Staff

Shawn Kalloway
Core Supervisor
(414) 937-4641 | (414) 937-6284 (fax)
Specialization: Zygote injections, Blastocyst infections, rederivation, cryopreservation, IVF

Jamie Foeckler
Sr. Research Technician
(414) 937-3889 | (414) 937-6284 (fax)
Specialization: Zygote injections, Blastocyst injections, gene targeting, rederivation, cryopreservation
Services
Resources
Transgenic Core Facility Ordering Information
- Approved Animal Protocol number for transgenic / knock-out mice, including available housing space
- Completed Order Form (this link jumps down to the order forms)
- Service Agreement Form (for On-Campus Investigators). A separate Service Agreement form will be emailed to off-campus Investigators.
- Completed "Condition of Use" form if using gene targeting/blastocyst injection service
- Documentation about experimental materials provided to the Core
- Documentation about established screening procedure to characterize transgenic offspring or ES cells.
The project will then be assigned a project ID, and placed in the work queue. Upon initiation of the experiment, you will receive regular updates about the project progress. At project completion, the Investigator will receive a summary and a copy of our laboratory notes.
The following forms may be filled out online and emailed directly to the Core Supervisor. If you are having problems emailing them, please print them out and fax them to the number listed below. If you have any questions or concerns, please contact the TC Core Supervisor.
- Blank Project Request (PDF)
- Blastocyst Injection Request (PDF)
- Conditions of Use Statement (PDF)
- E14Tg2a.4 ES Cells Experiment (PDF)
- Gene Targeting Request (PDF)
- Mouse Line Rederivation Request(PDF)
- Sperm Cryopreservation Request (PDF)
- Zinc Finger Injection Request (PDF)
- Zygote Microinjection Request (PDF)
Fees
The Core is not a business, and does not attempt to generate net revenue. The Core’s only mission is to support and enable experiments by other Investigators. Fees are levied to cover approximately 50% of the actual cost involved in performing these experiments. The production of transgenic or knock-out animals is a long-term, multistep process, and in most cases the cost of generating the mice is only a fraction of the total project expense. We strongly recommend taking these considerations into account when applying for funding. The fee for individual services depends on the scope of the project. For details, please contact the Core Director.