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- [1] Perfectly matched 20-nucleotide guide RNA sequences enable robust genome editing using high-fidelity SpCas9 nucleasesGenome Biology, 18Dingbo ZhangHuawei ZhangTingdong LiKunling ChenJin-Long QiuCaixia Gao
Perfectly matched 20-nucleotide guide RNA sequences enable robust genome editing using high-fidelity SpCas9 nucleases
被引：97
|作者：
Zhang, Dingbo
[1
,2
,3
]
Zhang, Huawei
[1
,2
]
Li, Tingdong
[1
,2
,3
]
Chen, Kunling
[1
,2
]
Qiu, Jin-Long
[4
]
Gao, Caixia
[1
,2
]
机构：
[1] Chinese Acad Sci, Inst Genet & Dev Biol, State Key Lab Plant Cell & Chromosome Engn, Beijing, Peoples R China
[2] Chinese Acad Sci, Inst Genet & Dev Biol, Ctr Genome Editing, Beijing, Peoples R China
[3] Univ Chinese Acad Sci, Beijing, Peoples R China
[4] Chinese Acad Sci, Inst Microbiol, State Key Lab Plant Genom, Beijing, Peoples R China
来源：
基金：
中国国家自然科学基金;
关键词：
CRISPR-CAS NUCLEASES;
HUMAN-CELLS;
SPECIFICITY;
SYSTEM;
WHEAT;
ENDONUCLEASES;
VERSATILE;
CLEAVAGE;
PLANTS;
RICE;
D O I：
10.1186/s13059-017-1325-9
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：
High-fidelity SpCas9 variants (eSpCas9 and SpCas9-HF1) have been engineered to reduce off-target effects. We found that changes in guide RNA length induced significant reductions in the editing activities of SpCas9 variants in plant cells. Single guide RNAs harboring precise, perfectly matched 20-nucleotide guide sequences are necessary for high on-target editing activities of eSpCas9 and SpCas9-HF1. Precise 20-nucleotide guide sequences derived from tRNA sgRNA precursors enable robust on-target editing by these variants with enhanced specificity. Our work reveals an effective way of enhancing the use of the high-fidelity SpCas9 nucleases for efficient and precise genome engineering.
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页数：7
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