Building method of donor pig suitable for kidney xenotransplantation
A technology for xenotransplantation, construction method, applied in the field of construction suitable for kidney xenotransplantation donor pigs
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Embodiment 1
[0022] A construction method suitable for kidney xenotransplantation donor pigs, the specific operation process is carried out according to the following steps:
[0023] 1) Knockout of the three major antigen genes that cause immune rejection
[0024] On the basis of porcine fibroblast cell lines, three antigen genes that cause immune rejection, GGTA1, CMAH, and B4GalNT2, were knocked out using CRISPR / Cas9 gene editing technology;
[0025] 2) Site-specific insertion of three humanized modification genes of hCD55, hCD59, and hCD46
[0026] On the basis of the above-mentioned GTKO / CMAHKO / B4GalNT2KO three-gene modification, three humanized modification genes of hCD55, hCD59, and hCD46 were inserted at specific sites to construct porcine fibroblasts with GTKO / CMAHKO / B4GalNT2KO / hCD55 / hCD59 / hCD46 six-gene modification Tie;
[0027] 3) Somatic cell nuclear transfer and embryo transfer
[0028] Using somatic cell nuclear transfer technology, the screened GTKO / CMAHKO / B4GalNT2KO / hCD5...
Embodiment 2
[0031] A construction method suitable for kidney xenotransplantation donor pigs, the specific operation process is carried out according to the following steps:
[0032] 1) Knockout of the three major antigen genes that cause immune rejection
[0033] On the basis of porcine fibroblast cell lines, three antigen genes that cause immune rejection, GGTA1, CMAH, and B4GalNT2, were knocked out using CRISPR / Cas9 gene editing technology;
[0034] 2) Site-specific insertion of three humanized modification genes of hCD55, hCD59, and hCD46
[0035] On the basis of the above-mentioned GTKO / CMAHKO / B4GalNT2KO three-gene modification, three humanized modification genes of hCD55, hCD59, and hCD46 were inserted at specific sites to construct porcine fibroblasts with GTKO / CMAHKO / B4GalNT2KO / hCD55 / hCD59 / hCD46 six-gene modification Tie;
[0036] 3) Somatic cell nuclear transfer and embryo transfer
[0037] Using somatic cell nuclear transfer technology, the screened GTKO / CMAHKO / B4GalNT2KO / hCD5...
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