Transgenics

The Mouse Genetic Engineering Facility (MGEF) provides services to generate, cryopreserve, recover, and study transgenic mice to understand human biology and disease. The MGEF is fully equipped to make genetically-engineered mice and is staffed with skilled personnel with decades of transgenic experience. An overview of the core’s services is below. More detailed service descriptions can be found in the Services section.

Mouse Model Generation

CRISPR Gene Editing

The MGEF has generated dozens of new mouse lines with engineered indels, deletions, point mutations, and conditional alleles using CRISPR/Cas9 technology.

CRISPR services include:

  • Project consultation to discuss best approaches and considerations
  • Allele design
  • CRISPR reagent preparation and gRNA testing in mouse embryos
  • Embryo microinjection
  • Screening of founder mice
  • Breeding of founders to test for germline transmission
  • Sequencing of N1 heterozygous animals to verify correct gene editing
  • Off-target analysis
  • Development of an automated genotyping assay in conjunction with Transnetyx

Mouse Embryonic Stem (ES) Cells and Conditional Knockout Mice

The MGEF has extensive experience culturing C57BL/6N ES cell clones from the International Knockout Mouse Project repositories and generating tissue-specific knockout mice by blastocyst injection. The MGEF can also carry out gene targeting in 129 strain and 129/B6 hybrid mouse ES cells.

Cryopreservation and Recovery

Sperm vs. Embryo Cryopreservation

The MGEF provides services for both sperm and embryo cryopreservation. Sperm cryopreservation is usually the preferred option as it is quick and efficient and requires only two proven breeder male mice. Embryo cryopreservation requires significantly more mice but is the preferred option when the mouse line is on a non-standard genetic background or when multiple alleles need to be maintained. Contact Bill Shawlot to discuss your specific needs.

Cryo-related services include:

  • Sperm and embryo cryopreservation
  • Embryo cryo recovery
  • In vitrofertilization (IVF) to recover strains from repositories and other Universities
  • IVF speed expansion to rapidly ramp up transgenic colonies
  • Rederivation to SPF status
  • Assistance with shipping and receiving cryopreserved materials

Breeding and Colony Management

The MGEF leverages its extensive experience working with mice to provide husbandry and breeding options to enable researchers to manage their transgenic colonies better.

Services include:

  • Custom mouse breeding, weaning, and colony management
  • Animal identification and tail snips
  • Genotyping (through the Transnetyx TAGCenter kiosk near the MGEF lab in the ARC)
  • Collection of embryos for phenotypic analysis and molecular experiments, including RNA profiling and ATAC sequencing
  • Tamoxifen administration for conditional gene knockout studies