Sun Xingshen, Yan Ziying, Yi Yaling, Li Ziyi, Lei Diana, Rogers Christopher S, Chen Juan, Zhang Yulong, Welsh Michael J, Leno Gregory H, Engelhardt John F
Department of Anatomy and Cell Biology, University of Iowa Carver College of Medicine, Iowa City, Iowa 52242, USA.
J Clin Invest. 2008 Apr;118(4):1578-83. doi: 10.1172/JCI34599.
Somatic cell gene targeting combined with nuclear transfer cloning presents tremendous potential for the creation of new, large-animal models of human diseases. Mouse disease models often fail to reproduce human phenotypes, underscoring the need for the generation and study of alternative disease models. Mice deficient for CFTR have been poor models for cystic fibrosis (CF), lacking many aspects of human CF lung disease. In this study, we describe the production of a CFTR gene-deficient model in the domestic ferret using recombinant adeno-associated virus-mediated gene targeting in fibroblasts, followed by nuclear transfer cloning. As part of this approach, we developed a somatic cell rejuvenation protocol using serial nuclear transfer to produce live CFTR-deficient clones from senescent gene-targeted fibroblasts. We transferred 472 reconstructed embryos into 11 recipient jills and obtained 8 healthy male ferret clones heterozygous for a disruption in exon 10 of the CFTR gene. To our knowledge, this study represents the first description of genetically engineered ferrets and describes an approach that may be of substantial utility in modeling not only CF, but also other genetic diseases.
体细胞基因靶向与核移植克隆相结合，为创建人类疾病的新型大型动物模型带来了巨大潜力。小鼠疾病模型往往无法重现人类表型，这凸显了生成和研究替代疾病模型的必要性。缺乏CFTR的小鼠一直是囊性纤维化（CF）的不良模型，缺乏人类CF肺部疾病的许多特征。在本研究中，我们描述了在家猫中使用重组腺相关病毒介导的成纤维细胞基因靶向，随后进行核移植克隆，产生CFTR基因缺陷模型的过程。作为该方法的一部分，我们开发了一种体细胞年轻化方案，通过连续核移植从衰老的基因靶向成纤维细胞中产生活的CFTR缺陷克隆。我们将472个重构胚胎移植到11只受体母貂体内，获得了8只健康的雄性雪貂克隆，它们对于CFTR基因第10外显子中的破坏是杂合的。据我们所知，本研究首次描述了基因工程雪貂，并描述了一种不仅对CF建模，而且对其他遗传疾病建模可能具有重大实用价值的方法。
