Cloning of the transgenic pigs expressing human decay accelerating factor and N-acetylglucosaminyltransferase III.
The present paper describes production of cloned pigs from fibroblast cells of transgenic pigs expressing human decay accelerating factor (DAF, CD55) and N-acetylglucosaminyltransferase III (GnT-III) that remodels sugar-chain biosynthesis. Two nuclear transfer protocols were used: a two-step activation (TA) method and a delayed activation (DA) method. Enucleated in vitro-matured oocytes and donor cells were electrically fused in a calcium-containing medium by TA method or in a calcium-free medium by DA method, followed by electrical activation 1-1.5 h later, respectively. In vitro blastocyst formation rates of nuclear transferred embryos reconstructed by TA and DA method were 8% and 14%, respectively. As a result of embryo transfer of the reconstructed embryos made by each method into recipient pigs, both gave rise to cloned piglets. These cloned pigs expressed transgene as much as their nuclear donor cells. In conclusions, (1) pig cloning can be carried out by TA or DA nuclear transfer methods, (2) expression of transgenes can be maintained to cloned pigs from the nuclear donor cells derived from transgenic animals.
Fujimura
Tatsuya
T
Animal Engineering Research Institute, Midorigahara, Tsukuba, Ibaraki 300-2646, Japan. t.fujimura@nipponham.co.jp
Kurome
Mayuko
M
Murakami
Hiroshi
H
Takahagi
Yoichi
Y
Matsunami
Katsuyoshi
K
Shimanuki
Shinichi
S
Suzuki
Kohei
K
Miyagawa
Shuji
S
Shirakura
Ryota
R
Shigehisa
Tamotsu
T
Nagashima
Hiroshi
H
D000818:Animals; D030801:Animals, Genetically Modified; D001755:Blastocyst / Q000166:cytology*; D018958:CD55 Antigens / Q000235:genetics*; D019976:Cloning, Organism / Q000379:methods*; D004624:Embryo Transfer; D005260:Female; D017351:N-Acetylglucosaminyltransferases / Q000235:genetics*; D009865:Oocytes / Q000166:cytology*; D013552:Swine
D018958:CD55 Antigens; D017351:N-Acetylglucosaminyltransferases; C054599:beta-1,4-mannosyl-glycoprotein beta-1,4-N-acetylglucosaminyltransferase