Transgenic Core Facility

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 Health Sciences Laboratory Animal Services (HSLAS) facility.

Research Resource Identifier - RRID:SCR_019175

Services

Note: All requested strains must be on an approved Animal Use Protocol prior to commencement of work at the Transgenic Core Facility.

Mouse Sperm Cryopreservation

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 Embryo Cyropreservation

Like sperm cryopreservation, embryo cryopreservation is utilized to preserve your mouse lines. This can reduce expenditures and animal usage for a strain that is not currently needed or to prevent genetic drift in lines that are actively being bred. This is a good alternative to sperm cryopreservation when your strain has multiple independent transgenes or mutations. For this procedure, we request 10 females (3-8 weeks of age) and 10 males (over 8 weeks of age). This would provide approximately 200 embryos for cryopreservation however only a portion of these will be viable after thawing.

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.

Mouse Rederivation

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.

Equipment

• Hamilton Thorne Computer-Aided Sperm Analysis System
• Sutter Xenoworks Micromanipulator
• Sutter Xenoworks Digital Microinjector
• Nikon TE2000 Inverted Microscope
• Leica M165 Dissecting Microscope
• MINC Benchtop Incubator
• Sutter Micropipette Puller
• Fine Science Tools Bead Sterilizer
• Baker EdgeGUARD Clean Bench
• TMC Air Isolation Table

Transgenic Core User Committee

• Dr. Anastassia Voronova - Academic Lead of the Transgenic Core Facility, Department of Medical Genetics

• Dr. Matthew Macauley - Department of Chemistry
• Dr. Keith Fenrich - Faculty of Rehabilitation Medicine
• Dr. John Ussher - Faculty of Pharmacy & Pharmaceutical Sciences

• Dr. Serene Wohlgemuth - Manager, Transgenic Core Facility
• Dr. Richard Lehner - Vice-Dean, Research (Basic), FoMD
• Dr. Nathan Bosvik - Director, Health Sciences Laboratory Animal Services, FoMD
• Ms. Colleen Sunderland - Manager, Core Research Facilities, FoMD