Epigenome editing by a CRISPR-Cas9-based acetyltransferase activates genes from promoters and enhancers

被引:1253
作者
Hilton, Isaac B. [1 ,2 ]
D'Ippolito, Anthony M. [2 ,3 ]
Vockley, Christopher M. [2 ,4 ]
Thakore, Pratiksha I. [1 ,2 ]
Crawford, Gregory E. [2 ,5 ]
Reddy, Timothy E. [2 ,6 ]
Gersbach, Charles A. [1 ,2 ,7 ]
机构
[1] Duke Univ, Dept Biomed Engn, Durham, NC 27706 USA
[2] Duke Univ, Ctr Genom & Computat Biol, Durham, NC 27708 USA
[3] Duke Univ, Med Ctr, Univ Program Genet & Genom, Durham, NC USA
[4] Duke Univ, Med Ctr, Dept Cell Biol, Durham, NC 27710 USA
[5] Duke Univ, Med Ctr, Dept Pediat, Div Med Genet, Durham, NC 27710 USA
[6] Duke Univ, Med Ctr, Dept Biostat & Bioinformat, Durham, NC USA
[7] Duke Univ, Med Ctr, Dept Orthopaed Surg, Durham, NC USA
基金
美国国家科学基金会; 美国国家卫生研究院;
关键词
FINGER TRANSCRIPTION FACTORS; ENDOGENOUS GENES; IN-VIVO; TARGETING ENHANCERS; CHROMATIN; CRISPR; CAS9; EXPRESSION; SYSTEM; CELLS;
D O I
10.1038/nbt.3199
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
Technologies that enable targeted manipulation of epigenetic marks could be used to precisely control cell phenotype or interrogate the relationship between the epigenome and transcriptional control. Here we describe a programmable, CRISPR-Cas9-based acetyltransferase consisting of the nuclease-null dCas9 protein fused to the catalytic core of the human acetyltransferase p300. The fusion protein catalyzes acetylation of histone H3 lysine 27 at its target sites, leading to robust transcriptional activation of target genes from promoters and both proximal and distal enhancers. Gene activation by the targeted acetyltransferase was highly specific across the genome. In contrast to previous dCas9-based activators, the acetyltransferase activates genes from enhancer regions and with an individual guide RNA. We also show that the core p300 domain can be fused to other programmable DNA-binding proteins. These results support targeted acetylation as a causal mechanism of transactivation and provide a robust tool for manipulating gene regulation.
引用
收藏
页码:510 / U225
页数:10
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