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The Rosa26 locus on mouse chromosome 6 is often used to integrate transgenic structures to achieve ubiquitous or conditional gene expression in mice and is considered to be one of the ideal locations for knockin targeted genes. The DNA fragments placed at the Rosa26 site are very safe and stable. Using the classical gene targeting method, obtain a gene knockin mouse at Rosa26 always takes one year or more. With the application of CRISPR/Cas9-mediated knockin technology, large DNA fragments can be accurately and efficiently inserted into the Rosa26.
CRISPR/Cas9 PlatformCB, a professional gene-editing technology services company, has extensive experience in the application of CRISPR/Cas gene-editing technology and has successfully knocked out/knocked specific genes in >300 mice. We guarantee to provide you with at least 2 Founder mice or 3 F1 mice and complete them within 3-5 months.
Workflow of Conditional Knockout Mouse Services Using CRISPR and Cre/LoxP
Based on the genes you are interested in, we will provide you with a comprehensive and efficient point mutation strategy to maximize Cas9 efficiency, including gRNA design and synthesis (target the sequence of the Rosa26 locus), donor vector construction, PCR primers for genotypic identification.
Synthesize gRNA, Cas9 mRNA in vitro, construct donor vector.
gRNA, Cas9 mRNA and donor vector are simultaneously injected into single-cell mouse embryos. Typically, we will inject as many embryos as needed to fulfill the guarantee.
Transfer healthy embryos to alternatives mother to produce founder mice. The Rosa26 locus sequence of founders will be detected by PCR and sequencing to reveal the insertion of genes your interest.
According to your needs, we will decide whether or not to breed the founder mice. Normally, we will breed up 3 founders which will be mated with wildtype mice of matched genetic background to generate their offspring. Then we will select F1 mice bearing targeted gene insertion at Rosa26 locus allele by genotyping.
CRISPR/Cas9 PlatformCB is one of the leading gene editing companies specializing in offering set of services for CRISPR/Cas gene-editing models. We have extensive experience in manipulating animal embryos and been very successful in incorporating CRISPR-Cas9 technology into our program, acquiring the customer's consistent good comments. Tell us about the genes you are interested in and we will provide you with a custom CRISPR/Cas Large-Fragment Knockin mouse model service, from strategy design to F1 mice, and provide a final report detailing the test strategy and verification method. we will meet your specific requirements, and protect your science investment.