The Animal Modeling Core (AMC) provides comprehensive services for the design and generation of genetically engineered and experimentally induced mouse models. Our team works with investigators to select the most appropriate strategy based on the biological question, desired genetic modification, mouse strain, experimental timeline, and downstream applications.
Projects range from routine knockout and knock-in models to complex conditional alleles, disease-associated variants, large DNA insertions, transgenic models, and specialized genome-engineering strategies. The AMC also offers AAV-based approaches for both genome editing and direct in vivo manipulation of gene expression.
Project Design and Consultation
The AMC provides an initial consultation to discuss project goals and identify the most appropriate strategy for generating the desired mouse model. Considerations may include:
- Type and complexity of the desired genetic modification
- Selection of CRISPR-Cas or alternative genome-engineering strategies
- Mouse strain and genetic background
- Germline versus somatic approaches
- Model validation and genotyping strategies
- Breeding requirements and experimental timelines
Our team can provide support from initial model conception and sequence design through founder generation, molecular validation, and establishment of the desired mouse line.
CRISPR-Cas Genome Editing
The AMC routinely uses CRISPR-Cas genome editing to generate customized mouse models directly in embryos. Applications include:
- Gene knockouts
- Point mutations and disease-associated variants
- Small and large targeted knock-ins
- Epitope and fluorescent protein tags
- Reporter alleles
- Cre and Flp recombinase knock-ins
- Conditional alleles containing loxP or FRT sites
- Humanized sequences and other customized genomic modifications
CRISPR reagents and DNA repair templates can be introduced into mouse embryos using electroporation, microinjection, AAV-mediated delivery, or combinations of these approaches depending on the requirements of the project.
Complex and Targeted Knock-In Models
Targeted insertion of larger DNA sequences can be challenging using conventional embryo genome-editing methods. The AMC has expertise in developing and applying alternative donor-template and delivery strategies for complex knock-in projects.
Approaches may include long single-stranded or double-stranded DNA repair templates, AAV-packaged donor DNA, optimized embryo-stage targeting, and other customized strategies designed to improve targeted integration efficiency.
The optimal approach is selected individually for each project based on the size and sequence of the desired insertion, genomic locus, and intended model.
Conditional and Tissue-Specific Mouse Models
The AMC can design and generate conditional alleles for spatially or temporally controlled gene modification. Services include generation of floxed alleles, conditional knock-in models, recombinase-dependent reporter alleles, and other customized designs compatible with Cre- or Flp-based systems.
We can also assist investigators with model design and breeding strategies involving existing Cre, CreERT2, Flp, or other recombinase-driver mouse lines.
AAV-Mediated and Somatic Mouse Models
Not every experimental question requires generation of a permanent germline mouse line. The AMC can assist investigators with AAV-based strategies to modify gene expression directly in vivo.
Potential applications include:
- Gene overexpression
- Expression of disease-associated or engineered gene variants
- Reporter gene expression
- RNA interference and gene knockdown
- Evaluation of candidate therapeutic constructs
- Tissue- or cell type-directed expression using appropriate promoters and AAV serotypes
These approaches can provide a rapid and flexible alternative to traditional germline model generation and may also be useful for proof-of-concept studies prior to development of a permanent genetically engineered line.
The AMC can assist with experimental design, AAV construct design, selection of appropriate vectors and delivery strategies, and coordination of in vivo studies.
Transgenic Mouse Production
The AMC offers traditional transgenic mouse production through embryo microinjection of engineered DNA constructs. This approach can be used for projects requiring random genomic integration of transgenes or other applications for which targeted genome editing is not required.
The AMC can assist with construct design, embryo manipulation, founder identification, and molecular characterization.
Embryonic Stem Cell-Based Mouse Production
Mouse models can also be generated using genetically modified embryonic stem (ES) cells. The AMC can perform blastocyst injection of targeted ES cells to generate chimeric mice and can work with commercially available or investigator-provided ES cell clones.
ES cell-based approaches remain useful for selected complex alleles and for existing targeted ES cell resources.
Advanced Mouse Genome-Editing Technologies
AMC investigators have developed and optimized specialized approaches for challenging mouse genome-engineering projects, including AAV-mediated donor delivery with 2-cell embryo CRISPR-Cas9 electroporation and CRISPR-SWITCH for controlled monoallelic genome editing.
Explore our Technology and Innovation page to learn more about genome-editing strategies developed by the AMC.
Have a Challenging Mouse Model?
Some projects do not fit neatly into a standard knockout or knock-in workflow. The AMC routinely works with investigators to develop customized genome-engineering and animal-modeling strategies for technically challenging projects.
Investigators are encouraged to contact the AMC early in project development for a consultation to discuss feasibility, experimental design, estimated timelines, and potential approaches.