Developing ABEmax-NG with Precise Targeting and Expanded Editing Scope to Model Pathogenic Splice Site Mutations In Vivo

被引：27

作者：

Huang, Shisheng ^{[1
,2
]}

Liao, Zhaodi ^{[3
,4
]}

Li, Xiangyang ^{[1
,2
]}

Liu, Zhen ^{[3
]}

Li, Guanglei ^{[1
,5
]}

Li, Jianan ^{[1
,2
]}

Lu, Zongyang ^{[1
,2
]}

Zhang, Yu ^{[1
]}

Li, Xiajun ^{[1
]}

Ma, Xu ^{[5
]}

Sun, Qiang ^{[3
]}

Huang, Xingxu ^{[1
,6
,7
]}

机构：

[1] ShanghaiTech Univ, Sch Life Sci & Technol, Shanghai 201210, Peoples R China

[2] Univ Chinese Acad Sci, Beijing 100049, Peoples R China

[3] Chinese Acad Sci, Inst Neurosci, Key Lab Primate Neurobiol, Ctr Excellence Brain Sci & Intelligence Technol, Shanghai 200031, Peoples R China

[4] Univ Chinese Acad Sci, Coll Life Sci, Beijing 100049, Peoples R China

[5] Natl Res Inst Family Planning, 12 Dahuishi Rd, Beijing 100081, Peoples R China

[6] Chinese Acad Sci, Ctr Excellence Mol Cell Sci, Shanghai Inst Biochem & Cell Biol, Shanghai 200031, Peoples R China

[7] Univ Chinese Acad Sci, Shanghai 200031, Peoples R China

来源：

ISCIENCE | 2019年 / 15卷

基金：

中国国家自然科学基金;

关键词：

Biological Sciences; Biological Sciences Tools; Molecular Biology;

D O I：

10.1016/j.isci.2019.05.008

中图分类号：

O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];

学科分类号：

07 ; 0710 ; 09 ;

摘要：

RNA splicing is related to many human diseases; however, lack of efficient genetic approaches to modulate splicing has prevented us from dissecting their functions in human diseases. Recently developed base editors (BEs) offer a new strategy to modulate RNA splicing by converting conservative splice sites, but it is limited by the editing precision and scope. To overcome the limitations of currently available BE-based tools, we combined SpCas9-NG with ABE-max to generate a new BE, ABEmax-NG. We demonstrated that ABEmax-NG performed precise A.T to G.C conversion with an expanded scope, thus covering many more splicing sites. Taking advantage of this tool, we precisely achieved A.T to G.C conversion exactly at the splice sites. We further modeled pathogenic RNA splicing in vitro and in vivo. Taken together, we successfully generated a versatile tool suitable for precise and broad editing at the splice sites.

引用

页码：640 / +

页数：30

共 15 条

[1] Analysis of off-target effects of CRISPR/Cas-derived RNA-guided endonucleases and nickases [J].