Pigs provide a valuable large animal model for several diseases due to their similarity with humans in anatomy, physiology, genetics and drug metabolism. We recently generated a porcine model for TP53^R167H and KRAS^G12D driven hepatocellular carcinoma (HCC) by autologous liver implantation. Here we describe a streamlined approach for developing genetically tailored porcine HCC cells by CRISPR/Cas9 gene editing and isolation of homogenous genetically validated cell clones. The combination of CRISPR/Cas9 editing of HCC cells described herein with the orthotopic HCC model enables development of various porcine HCC models, each with a specific mutational profile. This allows modeling the effect of different driver mutation combinations on tumor progression and in vivo testing of novel targeted therapeutic approaches in a clinically relevant large animal model.

中文翻译：

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通过 CRISPR/Cas9 编辑生成基因定制的猪肝癌细胞。


由于猪在解剖学、生理学、遗传学和药物代谢方面与人类相似，因此为多种疾病提供了有价值的大型动物模型。我们最近通过自体肝脏移植生成了TP53 ^R167H和KRAS ^G12D驱动的肝细胞癌 (HCC) 的猪模型。在这里，我们描述了一种通过 CRISPR/Cas9 基因编辑和分离同质基因验证细胞克隆来开发基因定制猪 HCC 细胞的简化方法。本文所述的 HCC 细胞的 CRISPR/Cas9 编辑与原位 HCC 模型的组合能够开发各种猪 HCC 模型，每种模型都具有特定的突变谱。这使得可以模拟不同驱动突变组合对肿瘤进展的影响，并在临床相关的大型动物模型中对新型靶向治疗方法进行体内测试。