GEMM has extensive experience making genetically modified mice. Many of these are classic gain of function mouse models, but we also have a molecular biologist on staff who can generate more complex and/or larger constructs using BAC recombineering.
We frequently combine the CRISPR/Cas9 technology with IVF to introduce genetic alterations or genes directly onto a wide range of background strains, including B6J, B6N, NOD, FVB, already genetically modified mice and others. Please inquire.
The general workflow for creating a gain of function mouse is as follows:
Gain of function mouse model services
- Consultation- meet to discuss project goals. If you have already made the transgene and are ready to inject zygotes, we’d like to meet to learn more about your transgene and genotyping assay.
- vector design and construction- GEMM will propose a strategy to generate the desired construct for researcher review and approval prior to project start. Once design is approved, GEMM will create the DNA constructs. Please reach out to us at any point during the process to check in and see how the project is progressing. Instead of being charged at project completion, you will be charged regularly (usually once per month) for work completed during that time period.
- Vector preparation for microinjection.
- Single copy assay detection- prior to detection, a genotyping assay that is sensitive enough to detect a single copy of your promotor+gene in the mouse genome must be developed.
- Pronuclear injection-GEMM will generate embryos by natural mating or by IVF, inject your promoter+gene, and perform embryo transfers.
- Resulting pups will be genotyped to identify putative founders. Mice can be transferred to researcher at this stage or kept in our breeding program to establish germline transmission.