Jan 12, 2016

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

Nature Biotechnology
Gozde KorkmazReuven Agami

Abstract

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 ERα (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.

  • References38
  • Citations79

References

  • References38
  • Citations79

Citations

Mentioned in this Paper

TP53 gene
Genome
Estrogen receptor alpha, human
CRISPR-Cas Systems
ESR1
Genetic Screening (Procedure)
Genomic Screening
TP53
MCF-7 Cells
ESR1 gene

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