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Updated: May 6, 2026
Methylated DNA Immunoprecipitation
Published on: January 2, 2009
Hydroxymethylated DNA immunoprecipitation (hmeDIP)
Colm E Nestor1, Richard R Meehan
1MRC Human Genetics Unit, IGMM, University of Edinburgh, Western General Hospital, Edinburgh, UK.
Abstract:
5-hydroxymethylcytosine (5hmC) was recently identified as an abundant epigenetic mark in mammals. Subsequent research has implicated 5hmC in normal mammalian development and disease pathogenesis in humans. Many of the techniques commonly used to assay for canonical 5-methylcytosine (5mC) cannot distinguish between 5hmC and 5mC. The development of antibodies specific to 5hmC has allowed for specific enrichment of DNA fragments containing 5hmC. Hydroxymethylated DNA immunoprecipitation (hmeDIP) has become an invaluable tool for determining both locus-specific and genome-wide profiles of 5hmC in mammalian DNA. Here, we describe the use of hmeDIP to characterize the relative abundance of 5hmC at loci in mammalian DNA.
Insights
5-hydroxymethylcytosine (5hmC) is a key epigenetic mark in mammals. Hydroxymethylated DNA immunoprecipitation (hmeDIP) using 5hmC-specific antibodies enables precise profiling of this mark in DNA.
Area of Science:
- Epigenetics
- Molecular Biology
- Genomics
Background:
- 5-hydroxymethylcytosine (5hmC) is an abundant epigenetic modification in mammals.
- 5hmC plays roles in mammalian development and human diseases.
- Standard methods for 5-methylcytosine (5mC) detection cannot differentiate between 5mC and 5hmC.
Purpose of the Study:
- To describe the utility of hydroxymethylated DNA immunoprecipitation (hmeDIP) for 5hmC analysis.
- To enable accurate locus-specific and genome-wide profiling of 5hmC.
- To characterize the relative abundance of 5hmC at specific DNA loci.
Main Methods:
- Development and application of antibodies specific to 5hmC.
- Utilizing hydroxymethylated DNA immunoprecipitation (hmeDIP) for DNA enrichment.
- Characterization of 5hmC abundance at selected mammalian DNA loci.
Main Results:
- Specific antibodies allow for targeted enrichment of 5hmC-containing DNA fragments.
- hmeDIP is effective for both locus-specific and genome-wide 5hmC profiling.
- The relative abundance of 5hmC can be accurately determined using hmeDIP.
Conclusions:
- hmeDIP is a powerful tool for studying 5hmC.
- This method facilitates understanding of 5hmC's role in mammalian biology and disease.
- Accurate 5hmC profiling is crucial for epigenetic research.
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