Our goal is to help you build the mouse model that will best address your experimental questions. We are flexible and can provide both full and partial services.
Vector Construction Only
We design and build your custom DNA targeting vector. You then use it for experiments, or use your core facility for electroporation, microinjection and mouse husbandry steps to produce your custom mouse model. You may need to perform the screening of the embryonic stem (ES) cells and the mouse genotyping yourself, depending on the capabilities of your facility. Should your facility fail to successfully generate the targeted ES cells or germline transmission mice, your project can be easily completed by our company at your request.
Targeted ES Cell Line Generation Only
Positively targeted ES cells are a major milestone accomplishment when it comes to generating genetically modified mouse models. We design and build the targeting vector, electroporate it into our robust ES cells, and subsequently screen the ES cell clones by PCR and Southern Blotting. We also perform chromosome counting to ensure a high percent euploidy. The targeted clones can be microinjected into blastocysts at your core facility to generate chimeras and subsequently F1 mice. Depending on the service offerings at your facility you may have to perform the mouse genotyping yourself.
We guarantee the generation of positively targeted ES cell clones if all work is performed at our facility. This includes all troubleshooting, if necessary, at no additional cost.
Utilizing our FLP ES cell lines alleviates the process for deleting the Neomycin selection cassette, saving you at least 3 months of time, and valuable funding.
Starting with a Targeting Construct Provided by a Third Party
We have worked with numerous targeting constructs provided to us by clients who have either generated their own constructs or purchased them from a third party. We can electroporate such constructs into our robust ES cells and provide them to you for screening, or we can perform the screening at our company. If we identify positive clones via PCR and Southern Blotting analysis, we will guarantee germline transmission at our facility, injecting as many times as necessary to produce chimeras and germline confirmed F1 mice.
Starting with KOMP/EUCOMM Targeting Constructs or Targeted ES Cell Clones
The International Knockout Mouse Consortium (includes KOMP and EUCOMM) has created an important resource of targeting constructs, targeted ES cells, and recently, targeted mice. If the targeted mice have not been created yet for your gene of interest, we can use the targeting vector or the targeted ES cell clones from the consortium to generate the mice.
We have generated several targeted ES cell lines and targeted mice from KOMP/EUCOMM targeting vectors. We electroporate your chosen construct into our ES cells and screen the resulting clones using PCR and Southern Blotting. If no positive clones are identified, you have the option of terminating the project without incurring any cost. This essentially provides our clients with a “free shot” at obtaining targeted ES cell clones. Once positive clones are identified and confirmed via Southern Blotting, we guarantee germline transmission mice as long as positive clones are injected at our facility.
Protect your investment with ingenious’ guarantee of germline transmission. Search now for your KOMP/EUCOMM targeting vector.
We suggest purchasing 3-4 individual clones if possible and performing essential quality control steps to ensure the clones are properly targeted and have high percentage euploidy. We can provide karyotyping and screening services, or those steps can alternatively be completed by the client. Confirmed, high quality clones should be selected for blastocyst injections to generate chimeras, and subsequently the germline confirmed F1 mice.
ingenious provides mouse model service options to address any and all of your needs, but our services do not stop there. Once you receive your model, your next step will be to generate a cohort of mice to utilize for experiments. The generation of experimental cohorts can be challenging, requiring multiple matings, strategic genotyping, and calculated colony scale up. ingenious offers prediction tools and expertise for developing the cohorts you need, as well as cryopreservation services for protecting your investment. Read more about our post project support services.