Methods in Molecular Biology 2009-01-01

Cloning of ES cells and mice by nuclear transfer.

Sayaka Wakayama, Satoshi Kishigami, Teruhiko Wakayama

Index: Methods Mol. Biol. 530 , 251-65, (2009)

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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.