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Updated: Jun 20, 2026
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Generation of Genetically Modified Mice through the Microinjection of Oocytes
Published on: June 15, 2017
Generation of gene knockout mice by ES cell microinjection
Glenn Longenecker1, Ashok B Kulkarni
1Department of Health and Human Services, National Institute of Dental and Craniofacial Research, National Institutes of Health, Bethesda, Maryland, USA.
Current Protocols in Cell Biology
|September 5, 2009
Summary
This unit details methods for producing chimeric mice, essential for creating gene knockout mice. Protocols cover embryo collection, stem cell injection, and transfer, alongside supporting techniques for efficient mouse model generation.
Area of Science:
- Animal models
- Developmental biology
- Genetics
Background:
- Chimeric mice are crucial for generating genetically modified organisms, particularly for gene knockout studies.
- Efficient production of chimeric mice relies on precise embryonic stem cell manipulation and transfer techniques.
Purpose of the Study:
- To describe detailed protocols for producing chimeric mice for gene knockout mouse generation.
- To provide comprehensive support protocols for all stages of chimeric mouse production.
Main Methods:
- Collection and handling of blastocyst embryos.
- Injection of embryonic stem (ES) cells into blastocysts.
- Uterine transfer of injected blastocysts into pseudopregnant recipients.
- Superovulation of donor mice and preparation of recipients.
Main Results:
- Established protocols for successful chimeric mouse production.
- Included methods for fabricating specialized laboratory equipment (glass pipets).
- Provided troubleshooting tips for common issues encountered during the process.
Conclusions:
- The described protocols provide a robust framework for generating chimeric mice.
- These methods facilitate the efficient creation of gene knockout mouse models.
- The unit serves as a practical guide for researchers in mouse genetics and developmental biology.
