Nuclear Transfer and Cloning
Nuclear transfer (NT) and cloning offers a unique and powerful experimental tool to study the mechanisms of gene reprogramming, to establish embryonic stem cells from somatic cells, and to clone live offspring. The process of NT involves two different cells. The first are oocytes, which are enucleated and provide the cellular components required for gene reprogramming and early embryo development. The second are donor cells, and their nuclei are injected into the enucleated oocytes. The donor cell nuclei are reprogrammed in the oocyte cytoplasm. The reconstructed oocytes are then activated and will begin to cleave. The embryos now have a genome identical to the original donor cells. These embryos can be used for basic research or implanting into foster mothers to develop to term. The protocols for mouse NT were developed in 1998 with assistance of the piezo drill device, and in 2000 without assistance of the piezo drill device. In this chapter, a comprehensive update on the techniques and protocols employed to generate mice by NT cloning are summarized based on the latest publications.
Key wordsNuclear transfer Cloning Somatic cells Oocytes Oocyte activation Embryo transfer Mouse
The author would like to thank supervisors, Mrs. Jufeng Qian, Northwest Agriculture University; Dr. Mark Westhusin, Texas A&M University; Dr. Colin McKerlie, Toronto Centre for Phenogenomics; and Mr. Steven Sansing, Charles River Laboratory.
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