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Investigating 5-Hydroxymethylcytosine (5hmC): The State of the Art

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Functional Analysis of DNA and Chromatin

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1094))

Abstract

The discovery of 5-hydroxymethylcytosine (5hmC) as an abundant base in mammalian genomes has excited the field of epigenetics, and stimulated an intense period of research activity aimed at decoding its biological significance. However, initial research efforts were hampered by a lack of assays capable of specifically detecting 5hmC. Consequently, the last 3 years have seen the development of a plethora of new techniques designed to detect both global levels and locus-specific profiles of 5hmC in mammalian genomes. This research effort has culminated in the recent publication of two complementary techniques for quantitative, base-resolution mapping of 5hmC in mammalian genomes, the first true mammalian hydroxymethylomes. Here, we review the techniques currently available to researchers studying 5hmC, discuss their advantages and disadvantages, and explore the technical hurdles which remain to be overcome.

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Acknowledgements

We thank members of the Meehan lab for advice on the development and application of 5hmC protocols. Work in RM’s lab is supported by the Medical Research Council, by the BBSRC, and by the Innovative Medicines Initiative Joint Undertaking (IMI JU) under grant agreement number 115001 (MARCAR project). URL: http://www.imi-marcar.eu/.

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Authors and Affiliations

  1. MRC Human Genetics Unit, IGMM, University of Edinburgh, Western General Hospital, Edinburgh, UK

    Colm E. Nestor, James P. Reddington & Richard R. Meehan

  2. The Centre for Individualised Medication, Linkoping University Hospital, Linkoping University, Linkoping, Sweden

    Colm E. Nestor & Mikael Benson

Authors
  1. Colm E. Nestor
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  2. James P. Reddington
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  3. Mikael Benson
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  4. Richard R. Meehan
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Editors and Affiliations

  1. Department of Biology, Autonomous University of Madrid Faculty of Sciences, Madrid, Spain

    Juan C. Stockert

  2. Departamento de Biología, Facultad de Ciencias, Universidad Autónoma de Madrid, Madrid, Spain

    Jesús Espada

  3. Department of Biology, Autonomous University of Madrid Faculty of Sciences, Madrid, Spain

    Alfonso Blázquez-Castro

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Nestor, C.E., Reddington, J.P., Benson, M., Meehan, R.R. (2014). Investigating 5-Hydroxymethylcytosine (5hmC): The State of the Art. In: Stockert, J., Espada, J., Blázquez-Castro, A. (eds) Functional Analysis of DNA and Chromatin. Methods in Molecular Biology, vol 1094. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-706-8_19

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  • DOI: https://doi.org/10.1007/978-1-62703-706-8_19

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  • Publisher Name: Humana Press, Totowa, NJ

  • Print ISBN: 978-1-62703-705-1

  • Online ISBN: 978-1-62703-706-8

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