Highly specific epigenome editing by CRISPR-Cas9 repressors for silencing of distal regulatory elements

被引：710

作者：

Thakore, Pratiksha I. ^{[1
,2
]}

D'Ippolito, Anthony M. ^{[2
,3
]}

Song, Lingyun ^{[2
,4
]}

Safi, Alexias ^{[2
,4
]}

Shivakumar, Nishkala K. ^{[1
]}

Kabadi, Ami M. ^{[1
,2
]}

Reddy, Timothy E. ^{[2
,5
]}

Crawford, Gregory E. ^{[2
,4
]}

Gersbach, Charles A. ^{[1
,2
,6
]}

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, Univ Program Genet & Genom, Durham, NC USA

[4] Duke Univ, Med Ctr, Dept Pediat, Div Med Genet, Durham, NC 27710 USA

[5] Duke Univ, Med Ctr, Dept Biostat & Bioinformat, Durham, NC 27710 USA

[6] Duke Univ, Med Ctr, Dept Orthopaed Surg, Durham, NC 27710 USA

来源：

NATURE METHODS | 2015年 / 12卷 / 12期

基金：

美国国家卫生研究院; 美国国家科学基金会;

关键词：

GENOME-WIDE ANALYSIS; TRANSCRIPTION FACTORS; ENDOGENOUS GENES; GLOBIN GENES; DNA-BINDING; RNA; ACTIVATION; CHROMATIN; EXPRESSION; CAS9;

D O I：

10.1038/nmeth.3630

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

Epigenome editing with the CRISPR (clustered, regularly interspaced, short palindromic repeats)-Cas9 platform is a promising technology for modulating gene expression to direct cell phenotype and to dissect the causal epigenetic mechanisms of gene regulation. Fusions of nuclease-inactive dCas9 to the Kruppel-associated box (KRAB) repressor (dCas9-KRAB) can silence target gene expression, but the genome-wide specificity and the extent of heterochromatin formation catalyzed by dCas9-KRAB are not known. We targeted dCas9-KRAB to the HS2 enhancer, a distal regulatory element that orchestrates the expression of multiple globin genes, and observed highly specific induction of H3K9 trimethylation (H3K9me3) at the enhancer and decreased chromatin accessibility of both the enhancer and its promoter targets. Targeted epigenetic modification of HS2 silenced the expression of multiple globin genes, with minimal off-target changes in global gene expression. These results demonstrate that repression mediated by dCas9-KRAB is sufficiently specific to disrupt the activity of individual enhancers via local modification of the epigenome.

引用

页码：1143 / +

页数：9

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