Knockin Rat Model Generation Service Using CRISPR/Cas9 - Research
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Custom designed gene targeting for the generation of conditional knock-in/knock-out rat models using our proprietary rat 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 rat 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 rat blastocysts. The resulting chimeras are bred for germline transmission. Germline transmission will be confirmed by PCR genotyping analysis and rats heterozygous for the targeted allele are shipped to the customer.
Optional services include: Cre/FLP recombination, production of homozygous rats, colony production and phenotyping.
All our animals are housed in a facility that is accredited by the Association for Assessment and Accreditation of Laboratory Animal Care (AAALAC) International, has an Assurance on file with the Office of Laboratory Animal Welfare (OLAW), and is a registered research facility with the U.S. Department of Agriculture.
Service Milestones for the Generation of Conditional Knock-in/Knock-out Rat Models
- Gene targeting vector design, construction and sequencing
- Rat ES cell targeting, screening and expansion of positive rat ES cell clones
- Karyotyping of targeted rat ES cells
- Chimera production via blastcyst injection
- Chimera breeding for germline transmission
- Genotyping of offspring
- Transfer of heterozygous targeted rats to the customer
- Optional: breeding to generate homozygous rats
- Optional: breeding with Cre rats for transgene activation (knockin) or gene elimination (knockout)