Cre-dependent Cas9-expressing pigs enable efficient in vivo genome editing

• Published in: Genome research 2017-12, Vol.27 (12), p.2061-2071
• Main Authors: Wang, Kepin, Jin, Qin, Ruan, Degong, Yang, Yi, Liu, Qishuai, Wu, Han, Zhou, Zhiwei, Ouyang, Zhen, Liu, Zhaoming, Zhao, Yu, Zhao, Bentian, Zhang, Quanjun, Peng, Jiangyun, Lai, Chengdan, Fan, Nana, Liang, Yanhui, Lan, Ting, Li, Nan, Wang, Xiaoshan, Wang, Xinlu, Fan, Yong, Doevendans, Pieter A, Sluijter, Joost P G, Liu, Pentao, Li, Xiaoping, Lai, Liangxue
• Format: Article
• Language: English
• Subjects: Animals; Animals, Genetically Modified; Bacterial Proteins - genetics; CRISPR-Cas Systems - genetics; Endonucleases - genetics; Female; Fibroblasts - metabolism; Gene Editing - methods; Gene Rearrangement; Genes, Tumor Suppressor; Genome; Humans; Integrases - metabolism; Lung Neoplasms - genetics; Male; Method; Oncogenes; Swine; Swine, Miniature - genetics; Transcription Activator-Like Effector Nucleases; Transcriptional Activation
• ISSN: 1088-9051; 1549-5469
• DOI: 10.1101/gr.222521.117

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 ( , , , , and ) and activate one oncogene ( ), 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.