Functional genetic screens for enhancer elements in the human genome using CRISPR-Cas9

被引：313

作者：

Korkmaz, Gozde ^{[1
]}

Lopes, Rui ^{[1
]}

Ugalde, Alejandro P. ^{[1
]}

Nevedomskaya, Ekaterina ^{[2
]}

Han, Ruiqi ^{[1
]}

Myacheva, Ksenia ^{[1
]}

Zwart, Wilbert ^{[2
]}

Elkon, Ran ^{[1
]}

Agami, Reuven ^{[1
,3
]}

机构：

[1] Netherlands Canc Inst, Div Biol Stress Response, Amsterdam, Netherlands

[2] Netherlands Canc Inst, Div Mol Pathol, Amsterdam, Netherlands

[3] Rotterdam Univ, Erasmus MC, Rotterdam, Netherlands

来源：

NATURE BIOTECHNOLOGY | 2016年 / 34卷 / 02期

关键词：

HUMAN-CELLS; BREAST-CANCER; P53; TRANSCRIPTION; MECHANISMS; ALIGNMENT; THERAPY; BINDING; CAS9; RNAS;

D O I：

10.1038/nbt.3450

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：

Systematic identification of noncoding regulatory elements has, to date, mainly relied on large-scale reporter assays that do not reproduce endogenous conditions. We present two distinct CRISPR-Cas9 genetic screens to identify and characterize functional enhancers in their native context. Our strategy is to target Cas9 to transcription factor binding sites in enhancer regions. We identified several functional enhancer elements and characterized the role of two of them in mediating p53 (TP53) and ERa (ESR1) gene regulation. Moreover, we show that a genomic CRISPR-Cas9 tiling screen can precisely map functional domains within enhancer elements. Our approach expands the utility of CRISPR-Cas9 to elucidate the functions of the noncoding genome

引用

页码：192 / +

页数：10

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