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Whole genome functional characterization of RE1 silencers using a modified massively parallel reporter assay

ABSTRACT: Whole genome functional characterization of RE1 silencers using a modified massively parallel reporter assay

PROVIDER: PRJNA803523 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR17887700.fastq.gz	Fastqsanger.gz
	SRR17887701.fastq.gz	Fastqsanger.gz
	SRR17887702.fastq.gz	Fastqsanger.gz
	SRR17887703.fastq.gz	Fastqsanger.gz
	SRR17887704.fastq.gz	Fastqsanger.gz

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Whole genome functional characterization of RE1 silencers using a modified massively parallel reporter assay

Project description:Both upregulation and downregulation by cis-regulatory elements help establish precise gene expression. Our understanding of how elements repress transcriptional activity is far more limited than activating elements. To address this gap, we characterized RE1, a group of transcriptional silencers bound by REST, on a genome-wide scale using an optimized massively parallel reporter assay (MPRAduo). MPRAduo empirically defined a minimal binding strength of REST required by silencer (REST m-value), above which multiple cofactors colocalize and act to directly silence transcription. We identified 1,500 human variants that alter RE1 silencing and found their effect sizes are predictable when they overlap with REST binding sites above the m-value. In addition, we demonstrate that non-canonical REST binding motifs exhibit silencer function only if they precisely align two half sites with specific spacer length. Our results show mechanistic insights into RE1 silencer which allows us to predict its activity and effect of variants on RE1, providing a paradigm for performing genome-wide functional characterization transcription factors binding sites.

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