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 for your next project, whether it is a CRISPR knockout, gene mutation, or sequence insertion.
CRISPR Knockout & Knockin Model Types:
Cas9 makes targeted cuts in your gene of interest which are repaired by an error-prone DNA repair pathway. Mutations introduced by this process disrupt the gene, to create a knockout allele.
Nucleotide changes are introduced at a target site by combining CRISPR/Cas9 with a repair oligo. Introduce a disease-causing mutation or humanize a critical amino acid with this strategy.
Insert a new sequence at a single site, for example add an epitope tag to label and purify your protein of interest.
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 when making complex models.
At ingenious, we work with you to design and customize your genetically engineered animal models, including CRISPR knockout, point mutation, or sequence insertion models. We will meet your exact specifications using our CRISPR/Cas9 technologies.
The 3 Step ingenious process
for generating your custom mouse model:
1st Step: Strategy & Design
Selection of target site, assays, and oligonucleotides, as well as predicted off-target sites. Preparation of sgRNA and Cas9 mRNA.
2ND STEP: PRODUCTION OF FOUNDER MICE
Injection of sgRNA and Cas9 mRNA into fertilized embryos, to generate FO mutated candidate animals. Breeding and genotyping to identify FO animals with the desired mutation.
3RD STEP: BREEDING OF FOUNDERS TO OBTAIN F1 MICE
The new founders are bred with wildtype mice of matching strain background, and their offspring genotyped to identify F1 mice bearing the knockout or knockin allele.
FASTER FOUNDERS WITH CRISPR
CRISPR/Cas9 enables the creation of founder mice for your new line in less time and at a lower cost. 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 generated from those cells. CRISPR bypasses that first step and makes the knockout directly in embryos, so screening for potential founders can begin months earlier than if ES cells are used. Point mutations and sequence insertions can also be made using CRISPR, in addition to knockouts. Scientists at ingenious have added the CRISPR strategy to our large portfolio of methods for creating your next model. We’ll work with you to create an indispensible line for your research.
After You Receive Your CRISPR Mice
At ingenious, it is possible to combine 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 provides the expertise needed for cohort development, as well as cryopreservation services to protect your investment.
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“We found iTL’s service to be exceptionally efficient and very well managed. We generated several mouse lines with iTL, and every step for each project was performed quickly and precisely, without any delays or problems.”
-Santos Franco, PhD