Abstract
We have been able to develop a stable nuclear transfer (NT) method in the mouse, in which donor nuclei are directly injected into the oocyte using a piezo-actuated micromanipulator. Although the piezo unit is a complex tool, once mastered it is of great help not only in NT experiments, but also in almost all other forms of micromanipulation. Using this technique, embryonic stem (ntES) cell lines established from somatic cell nuclei can be generated relatively easily from a variety of mouse genotypes and cell types. Such ntES cells can be used not only for experimental models of human therapeutic cloning but also as a means of preserving mouse genomes instead of preserving germ cells. Here, we describe our most recent protocols for mouse cloning.
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Acknowledgments
This work was supported by grants for Scientific Research in Priority Areas and the Project for the Realization of Regenerative Medicine (research field: technical development of stem cell manipulation) to T.W. from the Ministry of Education, Culture, Sports, Science and Technology of Japan.
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Wakayama, S., Kishigami, S., Wakayama, T. (2009). Cloning of ES Cells and Mice by Nuclear Transfer. In: Wurst, W., Kühn, R. (eds) Gene Knockout Protocols. Methods in Molecular Biology, vol 530. Humana Press. https://doi.org/10.1007/978-1-59745-471-1_13
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DOI: https://doi.org/10.1007/978-1-59745-471-1_13
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