CRISPR/Cas9 Knockin Mouse    

The CRISPR/Cas9 system is a very versatile tool for a wide variety of genetic changes, including deletions, point mutations, and insertions. The Cas9 nuclease contains two active sites. When both sites are active, Cas9 binds to a guide RNA molecule and causes targeted double-strand breaks (DSBs) in genomic DNA. The CRISPR system can also be used due to the presence of donor DNA molecules homologous to the target region. Specific nucleotide modifications are introduced into the sequence of interest during homologous recombination. Compared to traditional methods, the CRISPR-Cas9 system has significant advantages, including lower cost, shorter timeline and the ability to simultaneously change multiple genes.

CRISPR/Cas9 Platform^CB is a professional genetic editing biotechnology company that provides transgenic mouse models for biomedical research and drug discovery. We will provide you with a one-stop-shop from experimental design strategies to successful mouse model generation. The CRISPR/Cas9 platform^CB offers CRISPR-based knock-in mice at a very competitive price and complete in 3-4 months. We guarantee delivery of at least 2 founders or 3 F1 animals for knockin.

Workflow of CRISPR/Cas9-mediated Knockin Mouse Services

• Strategy and design

Based on the gene sequence you are interested in, we will provide you with nuclease-mediated strategy services, including target site selection, gRNA design, donor DNA design, donor vector (if needed) and non-target site prediction.

• CRISPR vector construction

Synthesize and validate gRNA, prepare Cas9 mRNA. Construct a DNA vector and a donor vector (if needed).

• Injection into mouse embryos

gRAN designed for a specific site of the target gene, Cas9 mRNA and donor DNA 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 to reveal any insertions.

• 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 knockin alleles.

If you have a target sequence (<200bp), we will provide you with dna oligonucleotides as donor dna. This approach is generally used to insert marker stop regions for specific functional and loxp sequences in conditional insertion models. If want a larger sequence (>200bp), we will provide you with a donor vector. This approach will allow you to insert luminescent or fluorescent reporter genes at specific target locations.

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
• Efficient CRISPR strategy and protocol for large segment knockin
• 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
• Quantitative assessment of gene expression
• Study the role of transcriptional regulatory elements
• Building a specific human disease model
• Drug screening and development
• Preclinical safety and toxicity studies
• Verify drug targets and specificities
• Genomic verification of antibodies

CRISPR/Cas9 Platform^CB is dedicated to providing professional CRISPR/Cas9 gene editing services. We will provide you with custom CRISPR/Cas9 knockin mouse service from gRNA design to F1 mice and keep in touch with you to report on project progress in a timely manner. We have first-rate laboratories, standards-compliant animal feeding conditions, and experienced scientists. We will meet your specific requirements to protect your scientific investment.