Mouse Models from ES cells to CRISPR and ‘BAC’

Types of Mouse Models from GeneTargeting.comOur highly experienced construct design team can generate targeting vectors or nuclease editing materials specifically tailored to meet your scientific needs. At ingenious, we first evaluate potential design strategies for your gene of interest. We then decide with you which technology will achieve the best results. For your custom project, we can use CRISPR and gene targeting in ES cells, either alone or in combination, to guarantee the most reliable production of your germline confirmed mouse model. Below is a list of the types of mouse models we can generate, based on the different targeting methods available.

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CRISPR

CRISPR: Overview from GeneTargeting.comCRISPR gene editing is an exciting technology which aims to speed up mouse model generation. ingenious offers injection of CRISPR reagents into embryos, as well as CRISPR-assisted homologous recombination in ES cells. The best strategy to use is based upon a thorough evaluation of the type and complexity of the targeting request.

Knockouts

Knockout: Overview from GeneTargeting.comingenious started out in 1998 as one of a few pioneering companies that developed knockout mouse models for the research community. From conventional deletions, to conditional knockouts and beyond, we have developed expertise to target even the most challenging genes.

No Doubt Conditional Knockout: Overview from GeneTargeting.comIn addition to standard conditional knockouts, ingenious has developed its No Doubt Conditional KnockoutTM, a sophisticated model for determining when and where your target gene has been disrupted. Utilizing our proprietary split-GFP design, reporter expression is tied to the recombination event and the restoration of GFP. Our technology removes concerns about leakiness and limited or low expression from the local gene promoter, as well as provides a reliable visual cue to show that your target gene has been knocked out.

VBAC Large Scale Targeting: Overview from GeneTargeting.comDon’t let size limitations affect your design options. At ingenious, 200 kb is the new standard. Our proprietary BAC and fosmid technologies allow us to perform large scale gene targeting in a fraction of the time that is typically required for these types of projects.

Knockins

Knockin: Overview from GeneTargeting.comKnockins refer to the insertion of exogenous sequence, such as a point mutation, cDNA or reporter gene insertion, minigene insertion or Rosa26 knockin to express or overexpress a gene of interest. Humanized mouse models are also considered knockins. At ingenious, we provide all of these types of knockin strategies, and more, depending on your experimental goals.

TruHumanization

Humanized Mice: Overview from GeneTargeting.comOur proprietary TruHumanization technology enables us to insert large sequences, such as entire human genes into the mouse genome. This provides our clients with the advantage of retaining intronic sequences as well as promoter and enhancer sequences which are often important in regulating gene expression.

Rosa26 Targeted Overexpression

The Rosa26 locus is a well known “safe harbor locus” used for expressing transgenes in the mouse without causing a secondary phenotype. The use of this locus has become extremely popular within the mouse transgenic community. In addition to standard Rosa26 targeted models, ingenious offers enhanced options for improved targeted transgenics:

RapidRosa26: Overview from GeneTargeting.comOur Rapid Rosa26 targeting technology allows us to create these models quickly and at a reduced cost when compared to standard knockout models. By utilizing well-validated gene targeting strategies combined with our ready-to-go enhancing vectors, we deliver your expression cassette or cDNA to the Rosa26 locus in record time.

Inducible Rosa26 Express: Overview from GeneTargeting.comDeveloping Rosa26 mouse models with inducible expression capabilities can be quite complicated and costly, and require mating with transcriptional activator or silencer mouse lines that might not already exist, when tissue-specificity is desired. ingenious has taken this complicated process and simplified it. Our Inducible Rosa26-Express targeting systems contains a floxed stop cassette and all of the necessary Tet-On control components, enabling tissue-specificity and inducibility without the need for transactivator or silencer mouse lines.

Inducible/Reversible Gene Expression

FAST Mice: Overview from GeneTargeting.comThis exciting technology was licensed exclusively to ingenious by Columbia University in 2014. Demonstrated in Science, F.A.S.T. (Flexible Accelerated STOP TetO) technology is a versatile tool where a single targeting event yields a multi-purpose mouse line with the potential for five or more controllable functionalities from the same locus, just using mating, including global KO, conditional rescue, and inducible control of gene expression.

Transgenics

Transgenic Mice: Overview from GeneTargeting.comWhen considering the creation of a transgenic mouse line, one can choose a random integration approach via pronuclear injection, or a more precise and predictable gene targeted approach using a “safe harbor locus” such as Rosa26. Here we discuss advantages and disadvantages of both transgenic approaches.

Komp/EUCOMM

eucomm from GeneTargeting.comThe International Knockout Mouse Consortium (KOMP, EUCOMM) is an important resource for targeting constructs, targeted embryonic stem cells, and recently- targeted mice. If mice do not yet exist for a gene of interest, we can work with either the targeting vector or the targeted ES cell clones from the consortium to generate mice.

After You Receive Your Mice

Post Project Services: Overview from GeneTargeting.comingenious provides technology and mouse model design options to address any and all of your needs, but our services do not stop there. Once you receive your model, you will need to generate a cohort of mice to utilize for your experiments. This may require multiple mating steps, specific genotyping, and calculated colony scale up. ingenious’ post project support services provides the tools you need for cohort development, as well as cryopreservation services for protecting your investment.

Can’t find what you are looking for?

Contact Us: Overview from GeneTargeting.comWe can generate any kind of mouse model, following our business motto “You think it up, We knock it out (or in!)” Contact us today to discuss your ideas and we will work with you to design a custom tailored mouse model, for your specific research goals.