PMID- 23947662
OWN - NLM
STAT- MEDLINE
DCOM- 20140304
LR  - 20171116
IS  - 1532-2378 (Electronic)
IS  - 1049-5398 (Linking)
VI  - 24
IP  - 4
DP  - 2013
TI  - Nucleofection-mediated alpha1,3-galactosyltransferase gene inactivation and membrane 
      cofactor protein expression for pig-to-primate xenotransplantation.
PG  - 253-67
LID - 10.1080/10495398.2012.752741 [doi]
AB  - Xenotransplantation of pig organs into primates leads to hyperacute rejection 
      (HAR). Functional ablation of the pig alpha 1,3-galactosyltransferase (GalT) gene, 
      which abrogates expression of the Gal alpha 1-3Gal beta 1-4GlcNAc-R (Gal) antigen, which 
      inhibits HAR. However, antigens other than Gal may induce immunological rejection 
      by their cognate antibody responses. Ultimately, overexpression of complement 
      regulatory proteins reduces acute humoral rejection by non-Gal antibodies when 
      GalT is ablated. In this study, we developed a vector-based strategy for ablation 
      of GalT function and concurrent expression of membrane cofactor protein (MCP, 
      CD46). We constructed an MCP expression cassette (designated as MCP-IRESneo) and 
      inserted between the left and the right homologous arms to target exon 9 of the 
      GalT gene. Nucleofection of porcine ear skin fibroblasts using the U-023 and 
      V-013 programs resulted in high transfection efficiency and cell survival. We 
      identified 28 clones in which the MCP-IRESneo vector had been successfully 
      targeted to exon 9 of the GalT gene. Two of those clones, with apparent 
      morphologically mitotic fibroblast features were selected through long-term 
      culture. GalT gene expression was downregulated in these 2 clones. Importantly, 
      MCP was shown to be efficiently expressed at the cell surface and to efficiently 
      protect cell lysis against normal human complement serum attack in vitro.
FAU - Ko, Nayoung
AU  - Ko N
AD  - a Animal Biotechnology Division , National Institute of Animal Science , RDA , 
      Suwon , South Korea.
FAU - Lee, Jeong-Woong
AU  - Lee JW
FAU - Hwang, Seong Soo
AU  - Hwang SS
FAU - Kim, Bella
AU  - Kim B
FAU - Ock, Sun A
AU  - Ock SA
FAU - Lee, Sung-Soo
AU  - Lee SS
FAU - Im, Gi-Sun
AU  - Im GS
FAU - Kang, Man-Jong
AU  - Kang MJ
FAU - Park, Jin-Ki
AU  - Park JK
FAU - Oh, Sung Jong
AU  - Oh SJ
FAU - Oh, Keon Bong
AU  - Oh KB
LA  - eng
PT  - Journal Article
PL  - England
TA  - Anim Biotechnol
JT  - Animal biotechnology
JID - 9011409
RN  - 0 (Membrane Cofactor Protein)
RN  - EC 2.4.1.- (Galactosyltransferases)
RN  - EC 2.4.1.87 (N-acetyllactosaminide alpha-1,3-galactosyltransferase)
SB  - IM
MH  - Analysis of Variance
MH  - Animals
MH  - Cell Survival
MH  - Cells, Cultured
MH  - Down-Regulation
MH  - Fibroblasts
MH  - Galactosyltransferases/*genetics/metabolism
MH  - Gene Silencing
MH  - Genetic Vectors/genetics
MH  - Membrane Cofactor Protein/*genetics/metabolism
MH  - Polymerase Chain Reaction
MH  - Swine
MH  - Swine, Miniature
MH  - Transfection/*methods
MH  - Transplantation, Heterologous
EDAT- 2013/08/21 06:00
MHDA- 2014/03/05 06:00
CRDT- 2013/08/17 06:00
PHST- 2013/08/17 06:00 [entrez]
PHST- 2013/08/21 06:00 [pubmed]
PHST- 2014/03/05 06:00 [medline]
AID - 10.1080/10495398.2012.752741 [doi]
PST - ppublish
SO  - Anim Biotechnol. 2013;24(4):253-67. doi: 10.1080/10495398.2012.752741.