GENE MODIFIED CLONED PIGS

基因修饰克隆猪

• 批准号: 6754508
• 负责人: Yifan Dai
• 金额: $ 25.68万
• 依托单位: UNIVERSITY OF PITTSBURGH AT PITTSBURGH
• 依托单位国家: 美国
• 财政年份: 2003
• 资助国家: 美国
• 起止时间: 2003-06-01 至 2006-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6754508
• 关键词: clone cells; diabetes mellitus; disease /disorder model; galactose; gene expression; genetic manipulation; genetically modified animals; immunocytochemistry; laboratory mouse; laboratory rabbit; nonhuman therapy evaluation; pancreatic islet transplantation; surface antigens; swine; xenotransplantation

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.

该提案的最终目的是通过基因技术缓解器官短缺危机 对猪进行改造，以便将猪的组织和器官移植到人类身上（即 异种移植）的结果相当于同种异体移植所达到的结果 常规免疫抑制。实现这一目标的进展已被阻碍 高等灵长类动物受体的先天免疫反应，其目标是αGal表位 在猪表面表达，但在人体细胞中不表达。已经表征了完整的基因组 几种低等哺乳动物中αGT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 与 PPL Therapeutics 合作，首次产生重组猪细胞， 具有双 alpha1,3GT 等位基因敲除 (KO)。这些双 KO 细胞可用于体细胞 细胞核移植（克隆）以产生克隆双 KO 胎儿和仔猪。我们打算 充分表征重组胎儿的分子改变和表型品质 细胞，以及预期足月克隆仔猪的细胞、组织或器官。在 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了测试，该模型将允许预测先天性和适应性 αGal 阴性高等灵长类动物接受者（包括人类）产生的免疫反应 转基因猪异种移植物。