A Generic Tool for Transcription Factor Target Gene Discovery in Arabidopsis Cell Suspension Cultures Based on Tandem Chromatin Affinity Purification | Zendy

Aurine Verkest | Zendy; Thomas Abeel | Zendy; Ken S. Heyndrickx | Zendy; Jelle Van Leene | Zendy; Christa Lanz | Zendy; Eveline Van De Slijke | Zendy; Nancy De Winne | Zendy; Dominique Eeckhout | Zendy; Geert Persiau | Zendy; Frank Van Breusegem | Zendy; Dirk Inzé | Zendy; Klaas Vandepoele | Zendy; Geert De Jaeger | Zendy

Open Access

A Generic Tool for Transcription Factor Target Gene Discovery in Arabidopsis Cell Suspension Cultures Based on Tandem Chromatin Affinity Purification

Author(s) -

Aurine Verkest,

Thomas Abeel,

Ken S. Heyndrickx,

Jelle Van Leene,

Christa Lanz,

Eveline Van De Slijke,

Nancy De Winne,

Dominique Eeckhout,

Geert Persiau,

Frank Van Breusegem,

Dirk Inzé,

Klaas Vandepoele,

Geert De Jaeger

Publication year - 2014

Publication title -

plant physiology

Language(s) - English

Resource type - Journals

SCImago Journal Rank - 3.554

H-Index - 312

eISSN - 1532-2548

pISSN - 0032-0889

DOI - 10.1104/pp.113.229617

Subject(s) - chromatin immunoprecipitation , chromatin , tandem affinity purification , arabidopsis , biology , transcription factor , computational biology , chip sequencing , dna , gene , chia pet , immunoprecipitation , arabidopsis thaliana , promoter , gene expression , chromatin remodeling , genetics , affinity chromatography , biochemistry , mutant , enzyme

Genome-wide identification of transcription factor (TF) binding sites is pivotal to our understanding of gene expression regulation. Although much progress has been made in the determination of potential binding regions of proteins by chromatin immunoprecipitation, this method has some inherent limitations regarding DNA enrichment efficiency and antibody necessity. Here, we report an alternative strategy for assaying in vivo TF-DNA binding in Arabidopsis (Arabidopsis thaliana) cells by tandem chromatin affinity purification (TChAP). Evaluation of TChAP using the E2Fa TF and comparison with traditional chromatin immunoprecipitation and single chromatin affinity purification illustrates the suitability of TChAP and provides a resource for exploring the E2Fa transcriptional network. Integration with transcriptome, cis-regulatory element, functional enrichment, and coexpression network analyses demonstrates the quality of the E2Fa TChAP sequencing data and validates the identification of new direct E2Fa targets. TChAP enhances both TF target mapping throughput, by circumventing issues related to antibody availability, and output, by improving DNA enrichment efficiency.

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