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Systematic mapping of functional enhancer-promoter connections with CRISPR interference
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Systematic mapping of functional enhancer-promoter connections with CRISPR interference
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Systematic mapping of functional enhancer-promoter connections with CRISPR interference
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Journal Article
Systematic mapping of functional enhancer-promoter connections with CRISPR interference
2016
Overview
Gene expression in mammals is regulated by noncoding elements that can affect physiology and disease, yet the functions and target genes of most noncoding elements remain unknown. We present a high-throughput approach that uses clustered regularly interspaced short palindromic repeats (CRISPR) interference (CRISPRi) to discover regulatory elements and identify their target genes. We assess >1 megabase of sequence in the vicinity of two essential transcription factors, MYC and GATA1, and identify nine distal enhancers that control gene expression and cellular proliferation. Quantitative features of chromatin state and chromosome conformation distinguish the seven enhancers that regulate MYC from other elements that do not, suggesting a strategy for predicting enhancer-promoter connectivity. This CRISPRi-based approach can be applied to dissect transcriptional networks and interpret the contributions of noncoding genetic variation to human disease.
Publisher
American Association for the Advancement of Science,The American Association for the Advancement of Science
Subject
/ Chromosome Mapping - methods
/ Clustered Regularly Interspaced Short Palindromic Repeats
/ Coding
/ Enhancer Elements, Genetic - genetics
/ Enhancer Elements, Genetic - physiology
/ GATA1 Transcription Factor - genetics
/ Genes
/ Genetics
/ High-Throughput Nucleotide Sequencing - methods
/ Humans
/ Joints
/ Mammals
/ Promoter Regions, Genetic - genetics
/ Promoter Regions, Genetic - physiology
/ Proteins
/ Proto-Oncogene Proteins c-myc - genetics
/ Real-Time Polymerase Chain Reaction
/ Screens
/ Strategy
