CRISPR Mouse Models
At the speed of CRISPR. Generate models quickly and efficiently.
ingenious is a proven leader in developing and deploying the latest technologies for creating gene targeted mouse models. Contact us today to find out how we can apply the power of CRISPR/Cas9 genome editing for your next project, whether it is a guide RNA-directed gene inactivation, modification or transgenic insertion.
CRISPR Knockout & Knockin Model Types
Cas9 generates targeted cuts in your gene of interest which are repaired by an error-prone DNA repair pathway, i.e., non-homologous endjoining (NHEJ). Mutations introduced by this process disrupt the gene either by nucleotide insertions or deletions (INDELs) to create a knockout allele.
Single nucleotide changes are introduced at a target site by combining CRISPR/Cas9 with a modified DNA template (e.g. an oligo) in order to support a repair pathway which utilizes homology-directed repair (HDR). Commonly this strategy applies to the modeling of disease-causing mutations or the humanization a critical amino acids.
New transgenic sequence components are inserted at a single, defined genomic site. This can entail the addition of an epitope tag to label and purify your protein of interest or can involve the insertion/replacement of original sequence with a transgene.
The CRISPR/Cas9 Process
The use of CRISPR/Cas9 to introduce targeted DNA modifications builds on decades of research into bacterial biology and DNA repair mechanisms. With this exciting new technology scientists at ingenious can generate mouse models at reduced cost in less time. Targeting materials are injected directly into embryos (bypassing the need to use ES cells) as part of a streamlined model production.
At ingenious, we work with you to design and customize your genetically engineered animal models, including CRISPR knockout, point mutation, or sequence insertion models and beyond. We will meet your exact specifications.
The 3 Step ingenious Process for Generating Your Custom Mouse Model
Selection of target site, assays, and oligonucleotides, as well as predicted off-target sites. Preparation of sgRNA and Cas9 mRNA. Injection of editing material into fertilized embryos to generate F0 mutated candidate animals resulting from embryo transfer into pseudopregnant mice. The new founders are bred with wildtype mice of matching strain background, and their offspring are genotyped to identify and select F1 mice bearing the intended knockout or knockin allele originating from the genetically heterogeneous (i.e., mosaic) F0 founder.
1st Step: Strategy and Design
2nd Step: Production of Founder Mice
3rd Step: Breeding of Founders to Obtain F1 Mice
Selection of target site, assays, and oligonucleotides, as well as predicted off-target sites. Preparation of sgRNA and Cas9 mRNA.
Injection of editing material into fertilized embryos to generate F0 mutated candidate animals resulting from embryo transfer into pseudopregnant mice.
The new founders are bred with wildtype mice of matching strain background, and their offspring are genotyped to identify and select F1 mice bearing the intended knockout or knockin allele originating from the genetically heterogeneous (i.e., mosaic) F0 founder.
- Faster Founders with CRISPR
CRISPR/Cas9 enables the creation of founder mice for your new line in less time and at a lower cost compared to cell-based approaches. Previously to make a knockout line the modification had to be made and verified in ES cells, which can take three months or more, then potential chimeric founders had to be generated from those cells. CRISPR eliminates this initial step and creates genetic changes directly in embryos, so that screening for potential founders can begin months earlier in relation to ES cell-derived animals. Scientists at ingenious have added CRISPR strategies to our large portfolio of methods for creating your next model. We are looking forward to working with you to create an indispensible line in support of your research.
- After You Receive Your CRISPR Mice
At ingenious, it is possible to breed CRISPR targeted models with additional mouse lines to generate experimental cohorts. This may require multiple mating steps, specific genotyping, and calculated colony scale up. ingenious’ post project support services provide the expertise needed for cohort development, as well as cryopreservation services to protect your scientific investment.