引导编辑：突破碱基编辑类型的新技术

doi:10.16288/j.yczz.22-156

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Hereditas(Beijing) ›› 2022, Vol. 44 ›› Issue (11): 993-1008.doi: 10.16288/j.yczz.22-156

• Review • Previous Articles Next Articles

Prime editing: a search and replace tool with versatile base changes

Yao Liu(), Xianhui Zhou, Shuhong Huang, Xiaolong Wang()

College of Animal Science and Technology/International Joint Research Center of Animal Bio-breeding, Ministry of Agriculture and Rural Affairs/Shaanxi Key Laboratory of Animal Genetic Breeding and Reproduction, Northwest A & F University, Yangling 712100, China

Received:2022-07-23 Revised:2022-09-01 Online:2022-11-20 Published:2022-09-16
Contact: Wang Xiaolong E-mail:yaoliu@nwafu.edu.cn;xiaolongwang@nwafu.edu.cn
Supported by:
the National Natural Science Foundation of China(31972526);the National Natural Science Foundation of China(32161143010)

Abstract

Abstract:

Prime editing is a newly developed CRISPR/Cas system-based genome editing technique. The effector of prime editor (PE) is termed PE2, which is generated by fusing a reverse transcriptase (RT) with a Cas9 H840A nickase. The guide RNA of PE is termed prime editing guide RNA (pegRNA), which consists of a single guide RNA (sgRNA) with a 3′ extension containing the RT template (RTT) and primer binding site (PBS). PE can install all 12 types of point mutations, small insertions and deletions and combinations thereof. Since its emergence in 2019, with the high versatility and specificity, PE has been applied to many living organisms, including animals, plants and bacteria. This led to many explorations of PE on gene therapy and genetic improvement in agriculture. In this review, we systematically describe the development, characteristics, optimizations, applications and security of PE. In addition, we discuss the future applications of PE. We expect that this review will help researchers to grasp and better use PE.

Key words: prime editor, CRISPR/Cas9, optimization, application

Yao Liu, Xianhui Zhou, Shuhong Huang, Xiaolong Wang. Prime editing: a search and replace tool with versatile base changes[J]. Hereditas(Beijing), 2022, 44(11): 993-1008.

Figures/Tables 6

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版本名称	优化策略	编辑效率及相应细胞	参考文献
ePE	抑制pegRNA环化	>80% (HEK293T)	[12]
epegRNA	抑制pegRNA降解	>80% (HEK293T)	[15]
xrPE	抑制pegRNA降解	>80% (HEK293T)	[13]
G-PE	抑制pegRNA降解	>70% (HEK293T)	[14]
spegRNA/apegRNA	RTT上引入同义突变/改变pegRNA二级结构	>80% (HEK293T)	[30]
—	增强pegRNA的表达	>50% (玉米细胞)	[16]
Dual-pegRNA	使用两条pegRNA来编码反向互补的逆转录产物	>40% (水稻细胞)	[32]
HOPE	使用成对的pegRNA	>30% (HCT116)	[17]
CMP-PE3	调节染色质结构	>47% (小鼠胚胎)	[19]
hyPE2	融合表达功能元件	>43% (HEK293T)	[18]
PE-P2/PE-P3	nCas9 N端融合RT，RTT上设计多碱基替换	>59% (水稻细胞)	[21]
PEmax	密码子优化，融合蛋白结构优化等	>30% (HEK293T)	[20]
PE2 *	增加核定位信号	>6% (小鼠肝脏)	[33]
ePPE	删除RT的RNase H结构域，添加病毒核衣壳蛋白	>25% (水稻细胞)	[22]
spCas9-NG等	利用识别不同PAM的Cas9变体构建PE系统	>50% (HEK293T)	[23]
Fn_PE3	利用FnCas9构建PE系统	>80% (HEK293FT)	[36]
PE4/PE5	调节DNA修复途径	>60% (HEK293T)	[20]
—	调节DNA修复途径	>40% (HEK293)	[25]

技术名称	改造策略	用途	Cas蛋白	PAM类型	参考文献
TwinPE	两条相向的pegRNA来逆转录出互补的DNA链	大片段的替换、插入或删除	nCas9	NGG	[37]
PRIME-Del	使用一对相向的pegRNA	精准删除大片段	nCas9	NGG	[38]
GRAND	使用一对相向的pegRNA	精准插入长片段	nCas9	NGG	[28]
PEn	Cas9和逆转录酶融合表达	小片段插入	Cas9	NGG	[24]
PEDAR	Cas9和逆转录酶融合表达	大片段删除与替换	Cas9	NGG	[29]
PEA1	Cas9和逆转录酶融合，多组分同一质粒表达	点突变、插入、删除	Cas9	NGG	[26]
WT-PE	Cas9和逆转录酶融合并结合成对pegRNA	大片段删除，染色体易位	Cas9	NGG	[27]

软件名称	特点	网址	参考文献
PrimeDesign	可进行高通量设计	http://primedesign.pinellolab.org/	[83]
PE-Designer	设计的同时展示潜在脱靶位点	http://www.rgenome.net/pe-designer/	[84]
Easy-Prime	基于机器学习优化pegRNA	http://easy-prime.cc/	[85]
PlantPegDesigner	优选双pegRNA策略	http://www.plantgenomeediting.net	[32]
pegIT	设计的同时展示潜在脱靶位点	https://pegit.giehmlab.dk	[86]
PINE-CONE	可进行高通量设计	https://github.com/xiaowanglab/PINE-CONE	[87]
PnB Designer	支持PE、CBE和ABE的gRNA设计	http://fgcz-shiny.uzh.ch/PnBDesigner/	[88]
pegFinder	可对pegRNA进行打分排序	http://pegfinder.sidichenlab.org	[89]

Prime editing: a search and replace tool with versatile base changes

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