Effective gene editing by high-fidelity base editor 2 in mouse zygotes

doi:10.1007/s13238-017-0418-2

Protein Cell

2017, Vol. 8

Issue (8) : 601-611 https://doi.org/10.1007/s13238-017-0418-2

RESEARCH ARTICLE

Effective gene editing by high-fidelity base editor 2 in mouse zygotes

Puping Liang^1,^2,³(

), Hongwei Sun¹, Ying Sun¹, Xiya Zhang¹, Xiaowei Xie¹, Jinran Zhang¹, Zhen Zhang^1,⁴, Yuxi Chen¹, Chenhui Ding³, Yuanyan Xiong¹, Wenbin Ma¹, Dan Liu⁵, Junjiu Huang^1,²(

), Zhou Songyang^1,^2,^3,⁵(

)

¹. Key Laboratory of Gene Engineering of the Ministry of Education, Guangzhou Key Laboratory of Healthy Aging Research and State Key Laboratory of Biocontrol, SYSU-BCM Joint Research Center, School of Life Sciences, Sun Yat-sen University, Guangzhou 510275, China
². State Key Laboratory of Ophthalmology, Zhongshan Ophthalmic Center, Sun Yat-sen University, Guangzhou 510275, China
³. Key Laboratory of Reproductive Medicine of Guangdong Province, School of Life Sciences and the First Affiliated Hospital, Sun Yat-sen University, Guangzhou 510275, China
⁴. Guangzhou Magigen Biotechnology Co.Ltd, Guangzhou 510320, China
⁵. Verna and Marrs Mclean Department of Biochemistry and Molecular Biology, Baylor College of Medicine, One Baylor Plaza, Houston, TX 77030, USA

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Abstract

Targeted point mutagenesis through homologous recombination has been widely used in genetic studies and holds considerable promise for repairing diseasecausing mutations in patients. However, problems such as mosaicism and low mutagenesis efficiency continue to pose challenges to clinical application of such approaches. Recently, a base editor (BE) system built on cytidine (C) deaminase and CRISPR/Cas9 technology was developed as an alternative method for targeted point mutagenesis in plant, yeast, and human cells. Base editors convert C in the deamination window to thymidine (T) efficiently, however, it remains unclear whether targeted base editing in mouse embryos is feasible. In this report, we generated a modified highfidelity version of base editor 2 (HF2-BE2), and investigated its base editing efficacy in mouse embryos. We found that HF2-BE2 could convert C to T efficiently, with up to 100% biallelic mutation efficiency in mouse embryos. Unlike BE3, HF2-BE2 could convert C to T on both the target and non-target strand, expanding the editing scope of base editors. Surprisingly, we found HF2-BE2 could also deaminate C that was proximal to the gRNA-binding region. Taken together, our work demonstrates the feasibility of generating point mutations in mouse by base editing, and underscores the need to carefully optimize base editing systems in order to eliminate proximal-site deamination.

Keywords base editor high-fidelity mouse embryos proximal-site deamination whole-genome sequencing

Corresponding Author(s): Puping Liang,Junjiu Huang,Zhou Songyang

Issue Date: 23 August 2017

Cite this article:

Puping Liang,Hongwei Sun,Ying Sun, et al. Effective gene editing by high-fidelity base editor 2 in mouse zygotes[J]. Protein Cell, 2017, 8(8): 601-611.

URL:

https://academic.hep.com.cn/pac/EN/10.1007/s13238-017-0418-2
https://academic.hep.com.cn/pac/EN/Y2017/V8/I8/601

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