Somatic cell nuclear transfer in the mouse.
Somatic cell nuclear transfer (SCNT) has become a unique and powerful tool for epigenetic reprogramming research and gene manipulation in animals since "Dolly," the first animal cloned from an adult cell was reported in 1997. Although the success rates of somatic cloning have been inefficient and the mechanism of reprogramming is still largely unknown, this technique has been proven to work in more than 10 mammalian species. Among them, the mouse provides the best model for both basic and applied research of somatic cloning because of its abounding genetic resources, rapid sexual maturity and propagation, minimal requirements for housing, etc. This chapter describes a basic protocol for mouse cloning using cumulus cells, the most popular cell type for NT, in which donor nuclei are directly injected into the oocyte using a piezo-actuated micromanipulator. In particular, we focus on a new, more efficient mouse cloning protocol using trichostatin A (TSA), a histone deacetylase (HDAC) inhibitor, which increases both in vitro and in vivo developmental rates from twofold to fivefold. This new method including TSA will be helpful to establish mouse cloning in many laboratories.
Kishigami
Satoshi
S
RIKEN, Center for Developmental Biology, Kobe, Japan.
Wakayama
Teruhiko
T
D000818:Animals; D002467:Cell Nucleus / Q000187:drug effects / Q000378:metabolism; D002478:Cells, Cultured; D002999:Clone Cells; D054885:Cumulus Cells / Q000166:cytology / Q000187:drug effects; D004622:Embryo, Mammalian / Q000187:drug effects; D005260:Female; D006877:Hydroxamic Acids / Q000494:pharmacology; D051379:Mice; D053652:Nuclear Transfer Techniques; D009865:Oocytes / Q000166:cytology / Q000187:drug effects / Q000378:metabolism
D006877:Hydroxamic Acids; C012589:trichostatin A