What We Do
The University of Alberta Transgenic Core Facility was
established to help researchers with their transgenic mouse production and
archiving needs. It is located in the
HSLAS animal facility.
Note: All requested strains must be on an approved Animal Use Protocol prior to commencement of work at the Transgenic Core Facility.
Sperm cryopreservation is an easy, inexpensive way to preserve your mouse lines of choice. Cryopreservation not only allows the researcher to stall breeding of lines not currently being used thereby reducing expenditure and animal usage and but it also prevents against disease (eg. pathogen outbreak) and genetic drift and allows for easier distribution of strains to other facilities. We request a single, proven-breeder male mouse (to ensure fertility), between 2-6 months of age. This provides enough sample for 8-10 straws to be frozen. Sperm will be checked for motility prior to and after cryopreservation. The cryopreservation service does NOT include an in vitro fertilization quality control step.
Mouse in vitro Fertilization
Mouse in vitro fertilization is usually used to recover strains that have been cryopreserved or to aid strains that have stopped breeding naturally. It is also used to rederive lines that have unknown health status or are known to have pathogens. Oocytes are collected from 5 superovulated females and incubated with sperm (either fresh or frozen) and cultured to the 2-cell stage after which the fertilized embryos are transferred to pseudopregnant CD1 females. The researcher will received all pups after weaning.
Rederivation of mice serves to remove pathogens from existing strains which may affect the researchers results and/or prevent them from maintaining the strain in the HSLAS facility, in the case of known excluded pathogens. To achieve this, an in vitro fertilization is performed using a male from the transgenic line of interest and wildtype commercially-available females. The resultant embryos are thoroughly washed to remove any potential contaminant and transferred to a pathogen-free female recipient. This female and any resultant pups will be housed separately to prevent cross-contamination into the HSLAS facility until health reports return confirming the animals are pathogen-free. The researcher will receive pups after this confirmation.
Transgenic Mouse Generation (DNA microinjection)
Transgenic mice are vital to many avenues of research, notably as a tool to model disease. The Transgenic Core Facility will inject your DNA construct into one of the pronuclei of newly fertilized oocytes and transfer the injected embryos into a pseudopregnant CD1 female. 3 round of this is performed (approximately 300 embryos injected). By default we use FVB animals but other strains may be used upon discussion. The researcher will received a tissue sample from each pups for genotyping and all identified founders.
CRISPR/Cas9-Mediated Genetically Engineered Mouse Production
The CRISPR/Cas9 technology has made the generation of new mouse lines with targeted mutations easier and more inexpensive than ever before. The CRISPR/Cas9 system can be used to make small insertions/deletions at random (via non-homologous end joining) or you can supply a donor template to make a targeted change (via homology-directed repair). Currently the user designs the guide RNA (and repair template, if required) but the Transgenic Core is able to offer advice on design. The CRISPR/Cas9 components are either electroporated or injected into newly fertilized embryos which are then transferred to a pseudopregnant CD1 female for gestation. At weaning the researcher is given a tissue sample from each pup that can be used for genotyping. When genotyping is complete, the desired pups are transferred to the PI.
- Hamilton Thorne Computer-Aided Sperm Analysis System
- Sutter Xenoworks Micromanipulator
- Sutter Xenoworks Digital Microinjector
- Nikon TE2000 inverted microscope
- MINC benchtop incubator
- Leica M165 dissecting microscope
Transgenic Core User Committee
- Dr. David Westaway - Academic Lead of the Transgenic Core Facility, Department of Medicine
- Dr. Basil Hubbard - Department of Pharmacology
- Dr. Gavin Oudit - Department of Medicine
- Dr. Anastassia Voronova, Department of Medical Genetics
- Dr. Serene Wohlgemuth - Manager, Transgenic Core Facility
Ex Officio Members
- Dr. Richard Lehner - Associate Dean, Research - Facilities and Cores, FoMD
- Dr. Wendy Magee - Director, Core Research Facilities, FoMD
- Dr. Nathan Bosvik - Director, Health Sciences Laboratory Animal Services, FoMD