GENE MODIFIED CLONED PIGS

这项建议的最终目标是通过基因治疗缓解器官短缺危机。 改变猪的组织和器官，以便将其移植到人类身上(即 异种移植)，其效果与同种异体移植相同 常规免疫抑制。实现这一目标的进展受阻于 较高灵长类受体的先天免疫反应，靶向于 在猪细胞表面表达，但不在人类细胞表面表达。已经确定了全基因组的特征 几种低等哺乳动物α-GT基因的组织和转录调控 (包括猪)和所有高等哺乳动物(包括人类)，我们正在合作 PPL Treateutics正在努力首次产生重组猪细胞， 有双α1，3GT等位基因敲除(KO)。这些双KO细胞可用于体细胞培养 细胞核移植(克隆)产生克隆的双KO胎儿和仔猪。我们打算 重组胎儿的分子改变和表型特征的充分表征 细胞，以及预期的足月克隆仔猪的细胞、组织或器官。在……里面 此外，双KO胎儿细胞，以及这些仔猪的细胞，组织和器官将被 在转基因小鼠模型中进行测试，这将允许预测先天和适应性 AlphaGal阴性的高级灵长类受体(包括人类)产生的免疫反应 转基因猪的异种移植物。

The ultimate objective of this proposal is to relieve the organ shortage crisis by genetic alteration of pigs so porcine tissues and organs can be transplanted to humans (i.e. xenotransplantation) with results equivalent to those attainable with allografts under conventional immunosuppression. Progress toward this objective has been precluded by the innate immune reaction of higher primate recipients that targets the alphaGal epitopes that are expressed on the surface of pig but not human cells. Having characterized the full genomic organization and transcriptional regulation of the alphaGT gene in several lower mammals (including pigs) and in all of the higher mammals (including humans), we are collaborating with PPL Therapeutics in their efforts to generate for the first time recombinant pig cells that have double alpha1,3GT allele knockout (KO). These double KO cells can be used for somatic cell nuclear transfer (cloning) to produce cloned double KO fetuses and piglets. We intend to fully characterize the molecular alterations and phenotypic qualities of the recombinant fetal cells, as well as the cells, tissues, or organs of the anticipated full-term cloned piglets. In addition, the double KO fetal cells, and the cells, tissues, and organs of these piglets will be tested in transgenic mouse models that will allow prediction of the innate and adaptive immune responses generated by alphaGal-negative higher primate recipients (including humans) to the genetically altered porcine xenografts.