Cre-dependent Cas9-expressing pigs enable efficient in vivo genome editing
- Kepin Wang1,2,3,10,
- Qin Jin1,2,3,10,
- Degong Ruan1,2,3,10,
- Yi Yang4,
- Qishuai Liu1,2,3,
- Han Wu1,2,3,
- Zhiwei Zhou1,2,3,
- Zhen Ouyang1,3,
- Zhaoming Liu1,3,
- Yu Zhao1,3,
- Bentian Zhao1,3,
- Quanjun Zhang1,3,
- Jiangyun Peng1,2,3,
- Chengdan Lai1,3,
- Nana Fan1,3,
- Yanhui Liang1,2,3,
- Ting Lan1,2,3,
- Nan Li1,2,3,
- Xiaoshan Wang1,2,3,
- Xinlu Wang5,
- Yong Fan4,
- Pieter A. Doevendans6,7,
- Joost P.G. Sluijter6,7,
- Pentao Liu8,
- Xiaoping Li1,3 and
- Liangxue Lai1,3,9
- 1CAS Key Laboratory of Regenerative Biology, Joint School of Life Sciences, Guangzhou Institutes of Biomedicine and Health, Chinese Academy of Sciences, Guangzhou Medical University, Guangzhou, 510530, China;
- 2University of Chinese Academy of Sciences, Beijing 100049, China;
- 3Guangdong Provincial Key Laboratory of Stem Cell and Regenerative Medicine, South China Institute for Stem Cell Biology and Regenerative Medicine, Guangzhou Institutes of Biomedicine and Health, Chinese Academy of Sciences, Guangzhou, 510530, China;
- 4Key Laboratory for Major Obstetric Diseases of Guangdong Province, Key Laboratory of Reproduction and Genetics of Guangdong Higher Education Institutes, The Third Affiliated Hospital of Guangzhou Medical University, Guangzhou 510150, China;
- 5Department of Nuclear Medicine, Guangzhou General Hospital of Guangzhou Military Command, Guangzhou 510010, China;
- 6Department of Cardiology, Experimental Cardiology Laboratory, University Medical Center Utrecht, Utrecht 3584CX, the Netherlands;
- 7Netherlands Heart Institute, Utrecht 3584CX, the Netherlands;
- 8Wellcome Trust Sanger Institute, Wellcome Trust Genome Campus Hinxton, Cambridge CB10 1HH, United Kingdom;
- 9Jilin Provincial Key Laboratory of Animal Embryo Engineering, Institute of Zoonosis, College of Veterinary Medicine, Jilin University, Changchun 130062, China
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↵10 These authors contributed equally to this work.
Abstract
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.
Footnotes
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[Supplemental material is available for this article.]
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Article published online before print. Article, supplemental material, and publication date are at http://www.genome.org/cgi/doi/10.1101/gr.222521.117.
- Received March 9, 2017.
- Accepted October 26, 2017.
This article is distributed exclusively by Cold Spring Harbor Laboratory Press for the first six months after the full-issue publication date (see http://genome.cshlp.org/site/misc/terms.xhtml). After six months, it is available under a Creative Commons License (Attribution-NonCommercial 4.0 International), as described at http://creativecommons.org/licenses/by-nc/4.0/.
