Conventional Knockout Mouse    

CRISPR / Cas9 is revolutionizing the field of mouse gene targeting. Mice have been very useful in the laboratory, and the first choice for studying mammalian biology. Methods obtaining knockout mice by ES cell homologous recombination often take a long time and are inefficient, and the success is not always guaranteed. As an emerging genetic editing technology, CRISPR/Cas9 can be used as a substitute for traditional gene-editing technology and has revolutionized the production of transgenic animals. This system shows unprecedented efficiency, reusability, and ease of use, reducing the time and cost of genome editing and enabling the production of animals with a wider range of genetic modifications.

CRISPR/Cas9 Platform^CB is a professional genetic editing biotechnology company that specializes in providing transgenic mouse models for biomedical research and drug discovery. CRISPR/Cas9 Platform^CB has independent experimental design strategies and verification methods, and successfully generated mouse models with fast turnaround and low cost. Based on our platform, we can knock out the targeted gene you interest in mice by CRISPR/Cas9 technology and complete in 3-5 months. We guarantee delivery of at least 2 founders or 3 F1 animals for the knockout.

Workflow of CRISPR/Cas9-mediated Knockout Mouse Services

• Strategy and design

Based on the genes you are interested in, we will provide you with a full range of experimental design services, including target site selection, gRNA design, and non-target site prediction.

• CRISPR vector construction

Synthesize and validate gRNA, prepare Cas9 mRNA.

• Injection into mouse embryos

gRAN designed for a specific site of the target gene and Cas9 are simultaneously injected into the mouse embryos. And we will inject as many embryos as needed to fulfill the guarantee.

• Production of founder mice

Embryos are transplanted into pseudopregnant mice to produce fonder mice. The young mice are verified and screened by PCR and sequencing analysis. A frameshift deletion/insertion of at least one allele in the gene of interest indicates that the knockout mouse is successfully constructed.

• Breeding of founders to obtain F1 mice

For some experimental subjects, it is sufficient to get a founder mouse. Based on your experimental needs, we will choose whether or not to breed the founder mice. The founder mice are mated with wild type mice of the same genetic background, and their progeny are genotyped to obtain F1 mice with knockout alleles.

When gRNA(s) designed for specific sites and Cas9 mRNA are simultaneously injected into fertilized eggs, cutting and incomplete repair at the target sites may result in gene insertion or deletion. If the cleavage site is located in the coding region of the gene, this may lead to frameshift mutation downstream of the site, resulting in conventional knockout. If exons in key coding domains need to be deleted, gRNAs and Cas9 of the upstream and downstream target sites of exons can be injected together to produce knockout mice with missing key regions.

Typically, we develop CRISPR-mediated knockout mice in C57BL/6 and FVB, but we can use other mouse strains according to your requirements.

Benefits

• High efficiency gRNA for ~100% Cas9 target cutting efficiency
• Obtain founder mice of your targeted gene in less time and at a lower cost
• F1 breeding to ensure germline transmission
• Timely report on project progress
• Convenient and efficient transportation service

Applications

• Study the biological functions of genes and proteins
• Building a specific human disease model
• Drug screening and development
• Preclinical safety and toxicity studies
• Verify drug targets and specificities
• Genomic verification of antibodies

At CRISPR/Cas9 Platform^CB, we will design and customize your CRISPR Knockout Animal Model for you. We will provide you with milestone progress and final report on the implementation of the project. And the final report details each step, including target vector design, construction and validation, transfection conditions, genotyping strategies and results. We will meet your specific requirements to protect your scientific investment. We look forward to working with you to create an indispensable support line for your research.