Genetically modified mice are commonly used as animal models of human diseases, providing insights that can lead to better understanding of disease processes, as well as for improved diagnosis and treatment.

The mouse is a powerful research tool in particular because of our ability to manipulate its genome directly. This provides a way to model specific human diseases when the causative gene or genes are known.

The Genome Engineering Core provides comprehensive genome engineering services that includes advice with strategy design and generation of new mouse models, characterisation of new or existing genetic alterations, vector targeting and/or ES cell tissue culture, and establishing founder mice.

The core provides MRL investigators with the following techniques:

ES-cell-based models

This ‘traditional’ gene-targeting method is particularly useful to generate complex mouse models. Mutant mice are generated by introducing mutations through homologous recombination in mouse embryonic stem (ES) cells. The core offers:

• advice on targeting constructs
• Purification and preparation of targeting constructs
• Screening ES cell clones
• Injection of  targeted ES cells into wild type blastocysts to contribute to the germline of chimeric mice
• Identification of chimeric mice and breeding to establish genetically modified mouse strains
• Breeding to establish and maintain genetically modified mouse strains

CRISPR-Cas9- based models

The core also uses nuclease based technologies such as  CRISPR/Cas9 which offer more efficient targeting and the possibility of multiplexed genome editing. The core provides:

• Design and preparation CRISPR/Cas9 reagents
• Microinjection of blastocysts (ES cell based projects) or of one-cell embryos with CRISPR/Cas9 reagents
• Genotyping to select for breeding to transmit the desired genome modification to their progeny.
• Breeding of F0 mice to establish genetically modified mouse lines

Models involving the International Knockout Mouse Consortium (IKMC)

The core also advises on vector targeting and/or ES-cell culture for projects incorporating material from publically available resources

such as EuCOMM, KOMP, NorCOMM and TIGM, includes vector targeting and/or ES cell tissue culture and general help with establishing founder mice.

 

We work closely with the CBS at the University of Cambridge for microinjection of one-cell embryos with CRISPR/Cas9 reagents or blastocyst injection (ES cell based projects)