ChIP-nexus enables improved detection of in vivo transcription factor binding footprints

Nature Biotechnology
Qiye HeJulia Zeitlinger

Abstract

Understanding how eukaryotic enhancers are bound and regulated by specific combinations of transcription factors is still a major challenge. To better map transcription factor binding genome-wide at nucleotide resolution in vivo, we have developed a robust ChIP-exo protocol called ChIP-nexus (chromatin immunoprecipitation experiments with nucleotide resolution through exonuclease, unique barcode and single ligation), which utilizes an efficient DNA self-circularization step during library preparation. Application of ChIP-nexus to four proteins--human TBP and Drosophila NFkB, Twist and Max--shows that it outperforms existing ChIP protocols in resolution and specificity, pinpoints relevant binding sites within enhancers containing multiple binding motifs, and allows for the analysis of in vivo binding specificities. Notably, we show that Max frequently interacts with DNA sequences next to its motif, and that this binding pattern correlates with local DNA-sequence features such as DNA shape. ChIP-nexus will be broadly applicable to the study of in vivo transcription factor binding specificity and its relationship to cis-regulatory changes in humans and model organisms.

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Datasets Mentioned

BETA
GSE55306
SRR770743
SRR770744

Methods Mentioned

BETA
immunoprecipitation
ChIP-seq
ChIP-exo
ChIP
iCLIP
Illumina sequencing
footprinting
PCR

Software Mentioned

DNA Shape
nexus
Amazon Web Services
Linux
R
ChIP
Peakzilla
bowtie
Twist
CASAVA

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