文献类型： 外文期刊
作者： Xu, Wen 1 ; Yang, Yongxing 1 ; Yang, Biying 1 ; Krueger, Christopher J. 3 ; Xiao, Qianlin 6 ; Zhao, Si 1 ; Zhang, Lu 1 ; Kang, Guiting 1 ; Wang, Feipeng 1 ; Yi, Hongmei 1 ; Ren, Wen 1 ; Li, Lu 1 ; He, Xiaoqing 1 ; Zhang, Chuanmao 2 ; Zhang, Bo 2 ; Zhao, Jiuran 1 ; Yang, Jinxiao 1 ;
作者机构： 1.Beijing Acad Agr & Forestry Sci, Beijing Key Lab Maize DNA Fingerprinting & Mol Br, Beijing, Peoples R China
2.Peking Univ, Coll Life Sci, Genome Editing Res Ctr, Key Lab Cell Proliferat & Differentiat,Minist Ed, Beijing, Peoples R China
3.Peking Univ, Coll Engn, Dept Biomed Engn, Beijing, Peoples R China
4.Georgia Inst Technol, Wallace H Coulter Dept Biomed Engn, Atlanta, GA 30332 USA
5.Emory Univ, Atlanta, GA 30322 USA
6.Southwest Univ, Coll Agron & Biotechnol, Chongqing, Peoples R China
期刊名称：NATURE PLANTS （ 影响因子：17.352； 五年影响因子：19.328 ）
ISSN： 2055-026X
年卷期： 2022 年 8 卷 1 期
页码：
收录情况： SCI
摘要： The ability to manipulate the genome in a programmable manner has illuminated biology and shown promise in plant breeding. Prime editing, a versatile gene-editing approach that directly writes new genetic information into a specified DNA site without requiring double-strand DNA breaks, suffers from low efficiency in plants(1-5). In this study, N-terminal reverse transcriptase-Cas9 nickase fusion performed better in rice than the commonly applied C-terminal fusion. In addition, introduction of multiple-nucleotide substitutions in the reverse transcriptase template stimulated prime editing with enhanced efficiency. By using these two methods synergistically, prime editing with an average editing frequency as high as 24.3% at 13 endogenous targets in rice transgenic plants, 6.2% at four targets in maize protoplasts and 12.5% in human cells was achieved, which is two- to threefold higher than the original editor, Prime Editor 3. Therefore, our optimized approach has potential to make more formerly non-editable target sites editable, and expands the scope and capabilities of prime editing in the future.
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