원문정보
Knock-in Somatic Cells of Human Decay Accelerating Factor and α1,2-Fucosyltransferase Gene on the α1,3-Galactosyltransferase Gene Locus of Miniature Pig
초록
영어
Galactose-α1,3-galactose (α1,3-Gal) epitope is synthesized at a high concentration on the surface of pig cells by α1,3-galactosyltransferase gene (GGTA1). The α1,3-Gal is responsible for hyperacute rejection in pig-to-human xenotransplantation. The generation of transgenic pigs as organ donors for humans is necessary to eliminate the GGTA1 gene that synthesize Galα(1,3)Gal. To prevent hyperacute graft rejection in pig-to-human xenotransplantation, previously, we developed α1,3-galactosyltransferase gene-knock-out somatic cell by homologous recombination. In this study, we established cell lines of α1,3-GT knock-out expressing hDAF and hHT gene from minipig fibroblasts to apply somatic cell nuclear transfer. The hDAF and hHT mRNA were expressed in the knock-in somatic cells and α1,3-GT mRNA was suppressed. However, the knock-in somatic cells were increased resistance to human serummediated cytolysis.
목차
재료 및 방법
α1,3-Galactosyltransferase 유전자 위치에서 hDAF와 HT유전자가 발현하는 knock-in vector 제작
미니돼지 체세포 배양
미니돼지 체세포에 knock-in vector 도입과 세포 선별
Knock-in된 체세포의 PCR 분석
RT-PCR과 Real-time PCR
Cytotoxicity assay
결과 및 고찰
α1,3-GT 유전자 위치에서 hDAF와 hHT 유전자가 발현하는 knock-in vector 구축
α1,3GT-hDAF-hHT knock-in vector 도입 및 선별
α1,3GT-hDAF-hHT knock-in 체세포에서 α1,3-GT, hDAF,hHT 유전자 발현 분석
α1,3GT-hDAF-hHT knock-in 체세포에서 human complementserum 처리에 의한 cytotoxicity 분석
요 약
REFERENCES