![]() ![]() ![]() Moreover, because of its high litter size, the cloning efficiency of Diannan miniature pigs was higher than those of 19 different donor cell types from other pigs. The Diannan miniature pig, a famous local variety, has unique advantages, including early sexual maturity, high birth rate and low full-grown body weight (compared with the Large White pig). Compared with non-human primates, miniature pigs present a decreased risk of cross-species disease transmission due to their greater phylogenetic distance from humans. Miniature pigs and humans have similar organ physiology and anatomy. Xenotransplantation (e.g., from pigs to humans) may resolve this problem. The number of organ donated cannot meet the demands of organ transplantation. The increasing life expectancy of humans has led to an increase in the number of patients suffering from chronic diseases and end-stage organ failure. Furthermore, FACS, confocal microscopy, IHC and western blotting analyses demonstrated that Gal epitopes were completely absent from the fibroblasts, kidneys and pancreas of one GTKO piglet. Sequencing analyses of the target site confirmed the homozygous GGTA1-null mutation in all fetuses and piglets, consistent with the genotype of the donor cells. In total, 12 live and two stillborn piglets were collected from six surrogates by either caesarean or natural birth. One live fetus was obtained from one surrogate by caesarean after 33 d of gestation for genotyping. Eight aborted fetuses were collected from the gilts that miscarried. The reconstructed GTKO embryos were subsequently transferred into 18 recipient gilts, of which 12 became pregnant, and six miscarried. The GGTA1 biallelic modified C99# cell line enabled high-quality SCNT, as evidenced by the 22.3 % (458/2068) blastocyst developmental rate of the reconstructed embryos. Three cell lines (3/126) showed GGTA1 biallelic knockout after modification by the TALENs. The luciferase SSA recombination assay revealed that the targeting activities of the designed TALENs were 17.1-fold higher than those of the control. ![]()
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