Techniques for Transgenic Mouse Production

Pronuclear Microinjection Platform

• Project design and discussion
• Generation of the targeting construct (optional)
• Injection of DNA into zygotes for production of founder mice
• Genotyping to identify F0 founders
• Mating of F0s to obtain germline-confirmed F1 mice

ES cell targeting Platform

• Project design and discussion
• Generation of targeting construct (optional)
• ES cell targeting and selection
• Screening and expansion of positive clones
• Microinjection of 2-3 positive clones into host blastocysts
• Chimera genotyping and breeding

CRISPR-Cas Platform

• Project strategy design and discussion
• Preparation of reagents based on specific project requirements
• Injection of CRISPR reagents into zygotes for production of founder mice
• Genotyping to identify F0 founders
• Mating of F0s to obtain germline-confirmed F1 mice