SpringerProtocols: Abstract: Cloning of ES Cells and Mice by Nuclear Transfer

Cloning of ES Cells and Mice by Nuclear Transfer

By: Sayaka Wakayama³, Satoshi Kishigami³, Teruhiko Wakayama³

Abstract

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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.

Affiliation(s): (3) Center for Developmental Biology RIKEN Kobe, Kobe, Japan

Book Title: Gene Knockout Protocols: Second Edition

Series: Methods in Molecular Biology | Volume: 530 | Pub. Date: Jun-01-2008 | Page Range: 251-265 | DOI: 10.1007/978-1-59745-471-1_13

Subject: Genetics/Genomics

Key Words: Nuclear transfer - reprogramming - ntES cell - cloning - embryo - stem cell - oocyte

References

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