High-efficiency prime editing with optimized, paired pegRNAs in plants

被引：256

作者：

Lin, Qiupeng ^{[1
,2
]}

Jin, Shuai ^{[1
,2
]}

Zong, Yuan ^{[1
]}

Yu, Hong ^{[3
,4
]}

Zhu, Zixu ^{[1
,2
]}

Liu, Guanwen ^{[2
,3
,5
]}

Kou, Liquan ^{[4
]}

Wang, Yanpeng ^{[1
,2
]}

Qiu, Jin-Long ^{[6
]}

Li, Jiayang ^{[2
,3
,4
]}

Gao, Caixia ^{[1
,2
]}

机构：

[1] Chinese Acad Sci, Innovat Acad Seed Design, Inst Genet & Dev Biol, State Key Lab Plant Cell & Chromosome Engn,Ctr Ge, Beijing, Peoples R China

[2] Univ Chinese Acad Sci, Coll Adv Agr Sci, Beijing, Peoples R China

[3] Chinese Acad Sci, Innovat Acad Seed Design, Inst Genet & Dev Biol, State Key Lab Plant Gen, Beijing, Peoples R China

[4] Chinese Acad Sci, Innovat Acad Seed Design, Inst Genet & Dev Biol, Natl Ctr Plant Gene Res, Beijing, Peoples R China

[5] Chinese Acad Sci, Inst Microbiol, State Key Lab Plant Genom, Beijing, Peoples R China

[6] Univ Chinese Acad Sci, CAS Ctr Excellence Biot Interact, Beijing, Peoples R China

来源：

NATURE BIOTECHNOLOGY | 2021年 / 39卷 / 08期

基金：

中国国家自然科学基金;

关键词：

GENES; RICE;

D O I：

10.1038/s41587-021-00868-w

中图分类号：

Q81 [生物工程学（生物技术）]; Q93 [微生物学];

学科分类号：

071005 ; 0836 ; 090102 ; 100705 ;

摘要：

Prime editing (PE) applications are limited by low editing efficiency. Here we show that designing prime binding sites with a melting temperature of 30 degrees C leads to optimal performance in rice and that using two prime editing guide (peg) RNAs in trans encoding the same edits substantially enhances PE efficiency. Together, these approaches boost PE efficiency from 2.9-fold to 17.4-fold. Optimal pegRNAs or pegRNA pairs can be designed with our web application, PlantPegDesigner. Improved guide RNAs enhance the efficiency of prime editing.

引用

页码：923 / +

页数：12

共 22 条

[1] Search-and-replace genome editing without double-strand breaks or donor DNA [J].