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FOXO Transcription Factors pp 115–130Cite as

Using ChIP-Based Approaches to Characterize FOXO Recruitment to its Target Promoters

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Part of the book series: Methods in Molecular Biology ((MIMB,volume 1890))

Abstract

Chromatin immunoprecipitation (ChIP) coupled to quantitative real-time PCR (ChIP-qPCR) or Next-Generation Sequencing (ChIP-seq) enables us to study the dynamics of chromatin recruitment of transcription factors (TFs). The popular model system Caenorhabditis elegans has provided us with fundamental understanding of the role of Insulin/IGF-1-like signaling (IIS) in metabolism and aging. The FOXO TF DAF-16 is the major output of the pathway that regulates most of the phenotypes associated with the IIS pathway. Here, we describe a ChIP protocol to study FOXO recruitment dynamics in whole C. elegans extracts. We discuss detailed practical procedures, including optimization, growth, harvesting, formaldehyde fixation, sonication of worms, TF immunoprecipitation for further downstream processing using qPCR as well as NGS for the analysis of FOXO-bound DNA.

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Author information

Authors and Affiliations

  1. Department of Reproductive Biology, All India Institute of Medical Sciences (AIIMS), New Delhi, India

    Neeraj Kumar

  2. Molecular Aging Laboratory, National Institute of Immunology, New Delhi, India

    Arnab Mukhopadhyay

Authors
  1. Neeraj Kumar
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  2. Arnab Mukhopadhyay
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Corresponding author

Correspondence to Arnab Mukhopadhyay .

Editor information

Editors and Affiliations

  1. Instituto de Investigaciones Biomédicas “Alberto Sols” (CSIC-UAM), Madrid, Spain

    Wolfgang Link

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Kumar, N., Mukhopadhyay, A. (2019). Using ChIP-Based Approaches to Characterize FOXO Recruitment to its Target Promoters. In: Link, W. (eds) FOXO Transcription Factors. Methods in Molecular Biology, vol 1890. Humana, New York, NY. https://doi.org/10.1007/978-1-4939-8900-3_10

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  • DOI: https://doi.org/10.1007/978-1-4939-8900-3_10

  • Published:

  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-4939-8899-0

  • Online ISBN: 978-1-4939-8900-3

  • eBook Packages: Springer Protocols

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