Efficient base editing for multiple genes and loci in pigs using base editors

Jingke Xie, Weikai Ge, Nan Li, Qishuai Liu, Fangbing Chen, Xiaoyu Yang, Xingyun Huang, Zhen Ouyang, Quanjun Zhang, Yu Zhao, Zhaoming Liu, Shixue Gou, Han Wu, Chengdan Lai, Nana Fan, Qin Jin, Hui Shi, Yanhui Liang, Ting Lan, Longquan Quan, Xiaoping Li, Kepin Wang () and Liangxue Lai ()
Additional contact information
Jingke Xie: Guangzhou Medical University
Weikai Ge: Guangzhou Medical University
Nan Li: Guangzhou Medical University
Qishuai Liu: Guangzhou Medical University
Fangbing Chen: Guangzhou Medical University
Xiaoyu Yang: Guangzhou Medical University
Xingyun Huang: Guangzhou Medical University
Zhen Ouyang: Guangzhou Medical University
Quanjun Zhang: Guangzhou Medical University
Yu Zhao: Guangzhou Medical University
Zhaoming Liu: Guangzhou Medical University
Shixue Gou: Guangzhou Medical University
Han Wu: Guangzhou Medical University
Chengdan Lai: Guangzhou Medical University
Nana Fan: Guangzhou Medical University
Qin Jin: Guangzhou Medical University
Hui Shi: Guangzhou Medical University
Yanhui Liang: Guangzhou Medical University
Ting Lan: Guangzhou Medical University
Longquan Quan: Guangzhou Medical University
Xiaoping Li: Guangzhou Medical University
Kepin Wang: Guangzhou Medical University
Liangxue Lai: Guangzhou Medical University

Nature Communications, 2019, vol. 10, issue 1, 1-13

Abstract: Abstract Cytosine base editors (CBEs) enable programmable C-to-T conversion without DNA double-stranded breaks and homology-directed repair in a variety of organisms, which exhibit great potential for agricultural and biomedical applications. However, all reported cases only involved C-to-T substitution at a single targeted genomic site. Whether C-to-T substitution is effective in multiple sites/loci has not been verified in large animals. Here, by using pigs, an important animal for agriculture and biomedicine, as the subjective animal, we showed that CBEs could efficiently induce C-to-T conversions at multiple sites/loci with the combination of three genes, including DMD, TYR, and LMNA, or RAG1, RAG2, and IL2RG, simultaneously, at the embryonic and cellular levels. CBEs also could disrupt genes (pol gene of porcine endogenous retrovirus) with dozens of copies by introducing multiple premature stop codons. With the CBEs, pigs carrying single gene or multiple gene point mutations were generated through embryo injection or nuclear transfer approach.

Date: 2019
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DOI: 10.1038/s41467-019-10421-8

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