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Cell-mediated transgenesis in rabbits: chimeric and nuclear transfer animals
V Zakhartchenko1, T Flisikowska, S Li
1Chair for Molecular Animal Breeding and Biotechnology, Ludwig-Maximilian University Muenchen, Munich, Oberschleissheim, Germany.
Biology of Reproduction
|October 15, 2010
Summary
Researchers achieved the first live cloned rabbits using genetically modified mesenchymal stem cells (MSCs). This breakthrough enables precise genetic engineering in rabbits for disease modeling in biomedical research.
Area of Science:
- Reproductive Biology
- Stem Cell Biology
- Genetics
Background:
- Precise genetic engineering in rabbits is crucial for creating disease models but hindered by challenges with embryonic stem cells and somatic cell nuclear transfer.
- Existing methods face limitations due to lack of functional rabbit embryonic stem cells and high mortality rates in somatic cell nuclear transfer.
Purpose of the Study:
- To investigate the potential of cultured pluripotent and multipotent cells for producing viable animals through nuclear transfer.
- To establish functional rabbit embryonic stem cells and mesenchymal stem cells (MSCs) for genetic engineering applications.
- To achieve the first live cloned rabbits from genetically manipulated cells.
Main Methods:
- Derivation and characterization of rabbit putative embryonic stem (ES) cells and mesenchymal stem cells (MSCs) from bone marrow.
- In vitro and in vivo assessment of pluripotent and multipotent differentiation capabilities.
- Somatic cell nuclear transfer using both ES cells and MSCs, followed by embryo development evaluation.
- Genetic manipulation of MSCs with fluorescent reporter genes and assessment of their nuclear transfer efficiency.
Main Results:
- Rabbit ES cells demonstrated in vitro and in vivo pluripotent differentiation, leading to the first ES-derived rabbit chimera.
- Rabbit MSCs showed successful multipotent differentiation and were more efficient nuclear donors than ES cells.
- Genetically modified MSCs yielded the first live cloned rabbits from manipulated cells, with similar efficiency to unmodified MSCs.
- Reporter gene expression in reconstructed embryos was not a reliable indicator of viability.
Conclusions:
- Mesenchymal stem cells are a viable and efficient cell source for nuclear transfer and genetic engineering in rabbits.
- The successful generation of cloned rabbits from genetically modified MSCs opens new avenues for creating genetically defined animal models.
- Further research is needed to refine methods for identifying viable embryos post-nuclear transfer.
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