TitleComparative isoschizomer profiling of cytosine methylation: the HELP assay.
Publication TypeJournal Article
Year of Publication2006
AuthorsKhulan, Batbayar, Thompson Reid F., Ye Kenny, Fazzari Melissa J., Suzuki Masako, Stasiek Edyta, Figueroa Maria E., Glass Jacob L., Chen Quan, Montagna Cristina, Hatchwell Eli, Selzer Rebecca R., Richmond Todd A., Green Roland D., Melnick Ari, and Greally John M.
JournalGenome Res
Date Published2006 Aug
KeywordsAnimals, CpG Islands, Cytosine, DNA Methylation, Genome, In Situ Hybridization, Fluorescence, Mice, Multigene Family, Nucleic Acid Hybridization, Promoter Regions, Genetic, Reverse Transcriptase Polymerase Chain Reaction

<p>The distribution of cytosine methylation in 6.2 Mb of the mouse genome was tested using cohybridization of genomic representations from a methylation-sensitive restriction enzyme and its methylation-insensitive isoschizomer. This assay, termed HELP (HpaII tiny fragment Enrichment by Ligation-mediated PCR), allows both intragenomic profiling and intergenomic comparisons of cytosine methylation. The intragenomic profile shows most of the genome to be contiguous methylated sequence with occasional clusters of hypomethylated loci, usually but not exclusively at promoters and CpG islands. Intergenomic comparison found marked differences in cytosine methylation between spermatogenic and brain cells, identifying 223 new candidate tissue-specific differentially methylated regions (T-DMRs). Bisulfite pyrosequencing confirmed the four candidates tested to be T-DMRs, while quantitative RT-PCR for two genes with T-DMRs located at their promoters showed the HELP data to be correlated with gene activity at these loci. The HELP assay is robust, quantitative, and accurate and is providing new insights into the distribution and dynamic nature of cytosine methylation in the genome.</p>

Alternate JournalGenome Res
PubMed ID16809668
PubMed Central IDPMC1524864
Grant ListR03 CA111577 / CA / NCI NIH HHS / United States