Despite being time-consuming and costly, generating genome-edited pigs holds great promise for agricultural, biomedical, and pharmaceutical applications. To further facilitate genome editing in pigs, we report here establishment of a pig line with Cre-inducible Cas9 expression that allows a variety of ex vivo genome editing in fibroblast cells including single- and multigene modifications, chromosome rearrangements, and efficient in vivo genetic modifications. As a proof of principle, we were able to simultaneously inactivate five tumor suppressor genes (TP53, PTEN, APC, BRCA1, and BRCA2) and activate one oncogene (KRAS), achieved by delivering Cre recombinase and sgRNAs, which caused rapid lung tumor development. The efficient genome editing shown here demonstrates that these pigs can serve as a powerful tool for dissecting in vivo gene functions and biological processes in a temporal manner and for streamlining the production of genome-edited pigs for disease modeling.

尽管费时且昂贵，但生成基因组编辑的猪在农业，生物医学和制药应用方面仍具有广阔的前景。为了进一步促进猪的基因组编辑，我们在这里报告建立具有Cre可诱导Cas9表达的猪系，该系允许在成纤维细胞中进行多种离体基因组编辑，包括单基因和多基因修饰，染色体重排以及有效的体内遗传修饰。作为原理的证明，我们能够同时使五个抑癌基因（TP53，PTEN，APC，BRCA1和BRCA2）失活，并激活一个癌基因（KRAS）。），这是通过递送Cre重组酶和sgRNA引起的，从而引起了肺肿瘤的快速发展。此处显示的有效的基因组编辑表明，这些猪可以作为一种有力的工具，以暂时的方式剖析体内的基因功能和生物学过程，并简化用于疾病建模的基因组编辑的猪的生产。