Enhancement of prime editing via xrRNA motif-joined pegRNA.

Zhang, Guiquan; Liu, Yao; Huang, Shisheng; Qu, Shiyuan; Cheng, Daolin; Yao, Yuan; Ji, Quanjiang; Wang, Xiaolong; Huang, Xingxu; Liu, Jianghuai

Zhang, Guiquan; Liu, Yao; Huang, Shisheng; Qu, Shiyuan; Cheng, Daolin; Yao, Yuan; Ji, Quanjiang; Wang, Xiaolong; Huang, Xingxu; Liu, Jianghuai.

Affiliation

Zhang G; State Key Laboratory of Pharmaceutical Biotechnology and MOE Key Laboratory of Model Animals for Disease Study, Model Animal Research Center at Medical School of Nanjing University, 210061, Nanjing, China.
Liu Y; Key Laboratory of Animal Genetics, Breeding and Reproduction of Shaanxi Province, College of Animal Science and Technology, Northwest A&F University, 712100, Yangling, Shaanxi, China.
Huang S; Gene Editing Center, School of Life Science and Technology, ShanghaiTech University, 100 Haike Rd., Pudong New Area, 201210, Shanghai, China.
Qu S; Gene Editing Center, School of Life Science and Technology, ShanghaiTech University, 100 Haike Rd., Pudong New Area, 201210, Shanghai, China.
Cheng D; State Key Laboratory of Pharmaceutical Biotechnology and MOE Key Laboratory of Model Animals for Disease Study, Model Animal Research Center at Medical School of Nanjing University, 210061, Nanjing, China.
Yao Y; Hangzhou Global Scientific and Technological Innovation Center, Zhejiang University, 311215, Hangzhou, China.
Ji Q; School of Physical Science and Technology, ShanghaiTech University, 100 Haike Rd., Pudong New Area, 201210, Shanghai, China.
Wang X; Key Laboratory of Animal Genetics, Breeding and Reproduction of Shaanxi Province, College of Animal Science and Technology, Northwest A&F University, 712100, Yangling, Shaanxi, China. xiaolongwang@nwafu.edu.cn.
Huang X; Gene Editing Center, School of Life Science and Technology, ShanghaiTech University, 100 Haike Rd., Pudong New Area, 201210, Shanghai, China. huangxx@shanghaitech.edu.cn.
Liu J; Zhejiang Laboratory, 311100, Hangzhou, China. huangxx@shanghaitech.edu.cn.

Nat Commun ; 13(1): 1856, 2022 04 06.

Article in En | MEDLINE | ID: mdl-35387980

ABSTRACT

ABSTRACT

The prime editors (PEs) have shown great promise for precise genome modification. However, their suboptimal efficiencies present a significant technical challenge. Here, by appending a viral exoribonuclease-resistant RNA motif (xrRNA) to the 3'-extended portion of pegRNAs for their increased resistance against degradation, we develop an upgraded PE platform (xrPE) with substantially enhanced editing efficiencies in multiple cell lines. A pan-target average enhancement of up to 3.1-, 4.5- and 2.5-fold in given cell types is observed for base conversions, small deletions, and small insertions, respectively. Additionally, xrPE exhibits comparable editindel ratios and similarly minimal off-target editing as the canonical PE3. Of note, parallel comparison of xrPE to the most recently developed epegRNA-based PE system shows their largely equivalent editing performances. Our study establishes a highly adaptable platform of improved PE that shall have broad implications.

Subject(s)

CRISPR-Cas Systems; Gene Editing; CRISPR-Cas Systems/genetics; Cell Line; Genome

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