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Overexpression transgenesis in mouse: pronuclear injection.
Wendy J K Gardiner1, Lydia Teboul
1Mary Lyon Centre, Medical Research Council, Harwell, UK.
Methods in Molecular Biology (Clifton, N.J.)
|June 9, 2009
Summary
This study details pronuclear injection protocols for creating transgenic animals. It covers DNA preparation, embryo production, and microinjection procedures for efficient genetic modification in mice.
Area of Science:
- Animal Biotechnology
- Molecular Biology
- Genetics
Background:
- Pronuclear injection is a key technique for generating transgenic animals.
- Efficient DNA delivery methods are crucial for successful genetic engineering.
Purpose of the Study:
- To provide comprehensive protocols for DNA preparation for microinjection.
- To describe the procedures for producing and handling mouse embryos for microinjection.
- To detail the microinjection process and embryo transfer into foster mothers.
Main Methods:
- Preparation of small and large plasmids, and BAC DNA for microinjection.
- Production of one-cell mouse embryos using specific breeding strategies.
- Microinjection of DNA into pronuclei of one-cell embryos.
- Surgical transfer of microinjected embryos into pseudo-pregnant foster females.
Main Results:
- Established protocols for preparing diverse DNA constructs (plasmids, BACs) for microinjection.
- Successfully produced and utilized one-cell mouse embryos for the procedure.
- Detailed the microinjection and embryo transfer steps, enabling efficient transgenic animal production.
Conclusions:
- The presented protocols offer a robust method for DNA delivery via pronuclear injection.
- This work facilitates the generation of transgenic mice for research purposes.
- Standardized procedures enhance the reproducibility and success rate of creating genetically modified animals.
