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Updated: Jul 19, 2026
08:48
Zygote Microinjection for Creating Gene Cassette Knock-in and Flox Alleles in Mice
Published on: June 24, 2022
Mouse zygotes as recipients in embryo cloning.
Pawel Greda1, Jolanta Karasiewicz, Jacek A Modlinski
1Department of Experimental Embryology, Institute of Genetics and Animal Breeding, Polish Academy of Sciences, Jastrze biec, 05-552 Wólka Kosowska, Poland.
Reproduction (Cambridge, England)
|October 31, 2006
Summary
A novel zygote enucleation technique allows early-stage nuclei transfer, enabling successful cloning. This method preserves essential factors for reprogramming introduced nuclei, leading to significant preimplantation and full-term development in cloned mice.
Area of Science:
- Reproductive Biology
- Developmental Biology
- Genetics
Background:
- Zygotes are typically not considered nuclear recipients for cloning.
- Previous enucleation methods failed to support blastocyst formation when using nuclei from later developmental stages.
Purpose of the Study:
- To develop a new zygote enucleation technique for successful nuclear transfer and cloning.
- To investigate the role of pronuclear factors in reprogramming introduced nuclei.
Main Methods:
- A selective zygote enucleation method was developed, removing only the pronuclear membrane and genetic material.
- Nuclei from eight-cell stage embryos were transferred into selectively enucleated zygotes.
- Cloned mouse development was assessed, and donor origin confirmed via coat color and glucose phosphate isomerase (GPI) isozyme analysis.
Main Results:
- Selective enucleation enabled preimplantation development in 70.5% of reconstructed zygotes.
- Full-term development was achieved in 7.8% of cases.
- Confirmation of cloned mouse origin from donor nuclei was successful.
Conclusions:
- The novel selective enucleation technique significantly improves the success rate of nuclear transfer in zygotes.
- Essential pronuclear factors, retained by selective enucleation, are crucial for reprogramming donor nuclei.
- This technique offers a promising advancement for cloning and developmental biology research.
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