RRC ID 1508
著者 Takahagi Y, Fujimura T, Miyagawa S, Nagashima H, Shigehisa T, Shirakura R, Murakami H.
タイトル Production of alpha 1,3-galactosyltransferase gene knockout pigs expressing both human decay-accelerating factor and N-acetylglucosaminyltransferase III.
ジャーナル Mol Reprod Dev
Abstract Heterozygous alpha 1,3-galactosyltransferase (GT) gene knockout pigs were produced with transgenic pig fetal cells expressing both human decay-accelerating factor (hDAF) and N-acetylglucosaminyltransferase III (GnT-III). In this study, we assessed the gene targeting efficiency in the transgenic pig fetal cells derived from different fetal tissues such as brain, skin, heart, and liver, or fetal carcass. Targeted cell colonies were selected by hygromycin B. The GT-knockout colonies (KO colonies) were obtained equally from the cells derived from all tissues except liver. Staining with five antibodies against intermediate filaments, all examined KO cell lines stained positive for vimentin with the exception of a colony that stained positive for both vimentin and glial fibrillary acidic protein simultaneously. This is the first study to produce KO cells from the astrocytes. Some of these KO cell lines were used for nuclear transfer (NT) to obtain KO pig fetuses. Fourteen fetuses were obtained from two recipients of the embryo transfer and eight of them had normal ploidy. The cells from the KO pig fetuses were also used for NT to produce cloned KO pigs. Two healthy clone pigs were born. These pigs were determined to have a heterozygous knockout GT gene and the two transgenes. The cells collected from the KO pigs were shown to have similar expression levels of hDAF and GnT-III compared to their original transgenic pigs and less than a half levels of the alphaGal epitopes existed in wild-type pig cells.
巻・号 71(3)
ページ 331-8
公開日 2005-7-1
DOI 10.1002/mrd.20305
PMID 15806557
MeSH Animals Animals, Genetically Modified / genetics* CD55 Antigens / biosynthesis CD55 Antigens / genetics* Cloning, Organism* Galactosyltransferases / deficiency* Gene Expression Humans N-Acetylglucosaminyltransferases / biosynthesis N-Acetylglucosaminyltransferases / genetics* Swine / genetics*
IF 2.823
引用数 80
WOS 分野 REPRODUCTIVE BIOLOGY BIOCHEMISTRY & MOLECULAR BIOLOGY DEVELOPMENTAL BIOLOGY CELL BIOLOGY
リソース情報
ヒト・動物細胞 NB1RGB(RCB0222)