Chaoran Zhao
Jun Xu
Yaxing Xu
Chunmei Cheng
Yinan Liu
Ting Wang
Yaqin Du
Liangfu Xie
Jingru Zhao
Yanchuang Han
Xiaobao Wang
Yun Bai
Hongkui Deng
1 Department of Cell Biology, School of Basic Medical Sciences, Peking University Stem Cell Research Center, State Key Laboratory of Natural and Biomimetic Drugs, Peking University Health Science Center and the MOE Key Laboratory of Cell Proliferation and Differentiation, College of Life Sciences, Peking-Tsinghua Center for Life Sciences, Peking University, Beijing 100191, China;
2 Shenzhen Stem Cell Engineering Laboratory, Key Laboratory of Chemical Genomics, Peking University Shenzhen Graduate School, Shenzhen 518055, China;
3 Peking University-Tsinghua University-National Institute of Biological Sciences Joint Graduate Program, College of Life Sciences, Peking University, Beijing 100871, China;
4 BeiHao Stem Cell and Regenerative Medicine Translational Research Institute, Beijing, China
Funds: This work was supported by the National Key Research and Development Program of China (2016YFA0100100 and 2017YFA0103000), the National Natural Science Foundation of China (Grant Nos. 31730059 and 31521004), the Guangdong Innovative and Entrepreneurial Research Team Program (2014ZT05S216), the Science and Technology Planning Project of Guangdong Province, China (2014B020226001 and 2016B030232001), the Science and Technology Program of Guangzhou, China (201508020001) and National Natural Science Foundation of China (Grant No. 31571052).
Received Date: 2018-03-14
Rev Recd Date:2018-05-16
Abstract
Abstract
One major strategy to generate genetically modified mouse models is gene targeting in mouse embryonic stem (ES) cells, which is used to produce gene-targeted mice for wide applications in biomedicine. However, a major bottleneck in this approach is that the robustness of germline transmission of gene-targeted ES cells can be significantly reduced by their genetic and epigenetic instability after long-term culturing, which impairs the efficiency and robustness of mouse model generation. Recently, we have established a new type of pluripotent cells termed extended pluripotent stem (EPS) cells, which have superior developmental potency and robust germline competence compared to conventional mouse ES cells. In this study, we demonstrate that mouse EPS cells well maintain developmental potency and genetic stability after long-term passage. Based on gene targeting in mouse EPS cells, we established a new approach to directly and rapidly generate gene-targeted mouse models through tetraploid complementation, which could be accomplished in approximately 2 months. Importantly, using this approach, we successfully constructed mouse models in which the human interleukin 3 (IL3) or interleukin 6 (IL6) gene was knocked into its corresponding locus in the mouse genome. Our study demonstrates the feasibility of using mouse EPS cells to rapidly generate mouse models by gene targeting, which have great application potential in biomedical research.Keywords: tetraploid complementation,
EPS,
mouse model,
CRISPR/Cas9
PDF全文下载地址:
http://www.protein-cell.org/article/exportPdf?id=a0a22ef7-8f68-4ef0-b1fd-bb046775467c&language=en
