Custom designed gene targeting for the generation of knock-in/knock-out mouse models using our proprietary129/C57BL6 hybrid ES cell line.
Our service includes consultation on available gene targeting vector design strategies and project assessment based on your model requirements.
Gene targeting vector is designed starting from a gene accession number. After construction of the targeting vector it is electroporated into mouse ES cells. Targeted ES cell clones are expand and further validated by genotyping, DNA sequencing and karyotyping to ensure optimal germline transmission efficiency. ES cell clones that passed QC requirements are injected into mouse blastocysts. The resulting chimeras are bred for germline transmission. Germline transmission will be confirmed by PCR genotyping analysis and mice heterozygous for the targeted allele are shipped to the customer.
Optional services include: Cre/FLP recombination, production of homozygous mice, colony production and phenotyping.
Our mice are housed in a facility that is accredited by the Association for Assessment and Accreditation of Laboratory Animal Care (AAALAC) International and is a registered research facility with the U.S. Department of Agriculture.
Service Milestones for the Generation of Knock-in and Knock-out Mouse Models
- Gene targeting vector construction and sequencing
- ES cell targeting, screening and expansion of positive ES cell clones
- Karyotyping of targeted ES cells
- Cre/FLP recombination (optional)
- Chimera production by blastocyst injection
- Breeding of chimeras for germline transmission
- Genotyping of offspring
- Transfer of heterozygous targeted mice to the customer
- Optional: breeding to generate homozygous mice