Back in 1998, ingenious started out as one of a few pioneering companies developing knockout mouse models. From conventional deletions to conditional knockouts and large scale deletions using BACs, we have developed the technology and expertise to target even the most challenging genes. Working with ingenious, our clients have comfort in knowing that they have chosen a company with longevity, backed by almost two decades of successful, published experience.
Utilizing both CRISPR and ES cell technologies, ingenious offers the following benefits for producing your knockout mouse model:
- Faster timelines (as short as 6 months) for germline-transmission knockout mouse models
- Guarantee delivery of your mouse model, not just your money back
- Cost effective utilizing our proprietary technologies
Below are some of ingenious’ core knockout specialties:
No-Doubt Conditional Knockout™: Certainty through GFP
Knockout your gene with certainty through GFP. By combining our split-GFP technology with our tried and true conditional targeting methodology, ingenious now offers the next generation of knockout mouse models. Our No-Doubt Conditional Knockout™ system provides strong GFP expression, independent of the endogenous gene promoter and in response to the Cre mediated recombination event, for straightforward identification of cells in which the gene deletion has occurred.
Large Scale Deletion
Base your designs around experiments, not limitations. Our proprietary BAC technology enables the specific deletion of very large genomic sequences, such as multiple genes or whole gene clusters, in a single targeting step. By innovating the design and construction of these once formidable targeting projects, ingenious has drastically reduced the timelines and costs associated with these knockout mouse models. At ingenious, 200 kb is the new standard.
CRISPR Assisted Conventional Knockout
Combining speeds normally associated with CRISPR/Cas9 and the reliability of traditional targeting via homologous recombination in ES cells, ingenious’ CRISPR assisted knockout models provide you with the best that both technologies have to offer, for creating targeted deletions. By utilizing the CRISPR/Cas9 system to increase targeting vector integration efficiency, we are able to reduce the time and cost associated with vector construction and ES cell screening by more than half, while still maintaining targeting quality.
Standard Conditional Knockout
The target region is flanked by loxP sites (“floxed”) to facilitate Cre recombinase-mediated excision, leading to tissue specific or temporal gene inactivation. Conditional knockouts have mostly replaced conventional knockouts nowadays as they no longer carry the risk of embryonic lethality. In addition, the Cre-loxP system allows researchers to develop much more versatile models. The parental floxed mouse line can be mated to different Cre or CreERT2 lines if desired. Global Cre expression will still allow for a total body knockout.
Standard Conventional Knockout
The target region to delete is replaced with a Neomycin selection cassette, or with a reporter gene. The targeted gene is thus inactivated at all times, in all tissues. Embryonic lethality is a possibility with conventional gene deletions. Because of this, many researchers now opt for conditional designs instead of conventional knockout mouse models.
After You Receive Your Mice
Breeding your knockout model with additional targeted models to generate experimental cohorts can be complex. ingenious’ post project support services aim to make things simple. Utilizing our proprietary prediction and modeling software, our team of breeding veterans build plans of action and generate the cohorts you need, when you need them and within projected timelines and costs.