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Chromatin Immunoprecipitation: Application to the Study of Asthma

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Molecular Genetics of Asthma

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1434))

Abstract

Chromatin immunoprecipitation (ChIP) is a technique for studying interactions between proteins and DNA in living cells. A protein of interest is selectively immunoprecipitated from a chromatin preparation, to analyze the DNA sequences involved. ChIP can be used to determine whether a transcription factor interacts with a candidate target gene and to map the localization of histones with posttranslational modifications on the genome.

The protein-DNA interactions are captured in vivo by chemical cross-linking. Cell lysis, DNA fragmentation, and immunoaffinity purification of the protein of interest allow to co-purify DNA fragments that are associated with that protein. The enriched protein-DNA population is ready to be quantified by PCR to detect precipitated DNA fragments. The combination of ChIP with DNA microarray analysis (ChIP-on-chip) and high-throughput sequencing (ChIP-seq) has enabled to obtain profiles of transcription factor occupancy sites and histone modifications throughout the genome.

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Acknowledgments

This work was supported by the project “Efecto del Ácido Retinóico en la enfermedad alérgica. Estudio transcripcional y su traslación a la clínica,” PI13/00564, integrated into the “Plan Estatal de I + D + I 2013–2016” and cofunded by the “ISCIII-Subdirección General de Evaluación y Fomento de la investigación” and the European Regional Development Fund (FEDER).

We are grateful to members of our laboratory for their stimulating discussion of this review.

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Authors and Affiliations

  1. Department of Biomedical and Diagnostic Sciences, University of Salamanca, Salamanca, Spain

    Asunción García-Sánchez

  2. Salamanca Institute for Biomedical Research (IBSAL), University Hospital of Salamanca, Salamanca, Spain

    Asunción García-Sánchez

  3. Salamanca Institute for Biomedical Research (IBSAL), Salamanca, Spain

    Fernándo Marqués-García

  4. Department of Clinical Biochemistry, University Hospital of Salamanca, Salamanca, Spain

    Fernándo Marqués-García

Authors
  1. Asunción García-Sánchez
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  2. Fernándo Marqués-García
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Corresponding author

Correspondence to Asunción García-Sánchez .

Editor information

Editors and Affiliations

  1. Department of Clinical Biochemistry, University Hospital of Salamanca, Salamanca, Spain

    María Isidoro García

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García-Sánchez, A., Marqués-García, F. (2016). Chromatin Immunoprecipitation: Application to the Study of Asthma. In: Isidoro García, M. (eds) Molecular Genetics of Asthma. Methods in Molecular Biology, vol 1434. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-3652-6_9

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  • DOI: https://doi.org/10.1007/978-1-4939-3652-6_9

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-3650-2

  • Online ISBN: 978-1-4939-3652-6

  • eBook Packages: Springer Protocols

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