UTSW Transgenic Technology Center

Overview of Services

The mission of the UTSW Transgenic Core is to create, propagate, and preserve lines of genetically engineered rodents. The Core staff generate conventional transgenic mice and rats from various inbred backgrounds using either single or multiple small transgenes and BACs. The Core has been creating genome edited rodents for ~ 6 years using the latest CRISPR/Cas9 editing tools. These efforts include creating simple KO’s, KI’s using oligo, megamer, or plasmid-based templates for homologous recombination (HR), and the creation of conditional alleles using either 2 oligo templates containing loxP motifs or a single long ssDNA template with the appropriate loxP motifs.  The Core staff continue to perform gene targeting in mouse embryonic stem (ES) cells on a small scale, expansion of in-house targeted ES cell clones and consortium-derived clones, and the preparation of select targeted ES cell clones for injection into blastocysts to create chimeric mice.

The Core staff routinely provide UTSW Investigators access to a wide array of assisted reproductive techniques. These include the cryopreservation of sperm, 2-cell embryos and ovaries from select lines of mice. The frozen gametes are stored in the Core’s cryopreservation facility on the South Campus and can be distributed to Investigators world-wide. Core staff can reconstitute lines of mice from cryopreserved gametes by either thawing embryos and transferring them to surrogate females or by thawing sperm, performing in vitro fertilization followed by embryo transfers. The Core staff routinely rederive lines of mice from conventional housing to an SPF barrier status. If the need arises, the Core can rescue a line of mice that has reached reproductive senescence or propagate a line of mice so that a very large number of animals can be produced in a single day.

Services Offered

• Generation of conventional transgenic mice and rats in inbred backgrounds
• Generation of simple KOs (INDEL based), complex KOs, simple KIs with oligo, long ssDNA or plasmid-based HR templates, and KIs with conditional alleles using either oligo or megamer HR templates
• Generation of ES cells harboring select mutation by gene targeting
• Expansion of ES cell clones generated in-house, acquired from a consortium or acquired from another Institution
• Generation of KO/KI mice by using genetically engineered ES cells to make chimeric mice
• Cryopreservation of gametes and ovaries
• Recovery of mouse lines from cryopreservation: Embryo thaw and transfer, IVF and embryo transfer, ovary transplantation
• Rederivation of lines of mice from conventional housing to a SPF barrier health status
• Rescue of senescent mouse lines by IVF and embryo transfer
• Propagation of mouse lines by IVF and embryo transfer

Protocols

• General Guidelines and Useful Tips for Design of Genome Editing Project
• Preparation of DNA for microinjection-small transgenes
• Preparation of BAC DNA for microinjection
• Genotyping of transgenic rodents
• Preparation of targeting vector DNA for electroporation
• Screening of 96-well plates for gene targeting
• Screening of 6-well plates to confirm gene targeting
• Breeding of chimeras to obtain germ line transmission: In-house ES cell clones and KOMP clones:  SM-1 derived Chimera Breeding  and KOMP clones Chimera breeding
• Fostering guidelines

Leadership

Robert Hammer, Ph.D. | Director 
robert.hammer@utsouthwestern.edu
214-645-6292

Location and hours of operation

Links and Resources

1. ISTT - Keep up to date with current research and breakthroughs in transgenic technologies.
2. MMRRC - Distributes and cryopreserves scientifically valuable, genetically engineered mouse strains and mouse ES cell lines.
3. UTSW Animal Resource Center - Location for animal policies, veterinary services, ordering, transports, and more

Contacts