A novel method to assess the full genome methylation profile using monoclonal antibody combined with the high throughput based microarray approach
A novel method to assess the full genome methylation profile using monoclonal antibody combined with the high throughput based microarray approach
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DOI：
10.4161/epi.4.6.9768
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发表时间：
2009-08
期刊：
影响因子：
3.7
通讯作者：
Ashwin Kelkar;D. Deobagkar
中科院分区：
文献类型：
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作者：
Ashwin Kelkar;D. Deobagkar
在这项工作中，我们展示了一种新的方法，利用免疫亲和检测杂交到cDNA微阵列的甲基化DNA的存在下的甲基化检测。我们使用一种特异于5甲基胞嘧啶的单克隆抗体来检测核型为45 XO的人成纤维细胞基因组DNA中5甲基胞嘧啶的存在。我们报告说，超过2900个基因在这种情况下显示甲基化的存在。我们还报告说，在这些实验的所有重复中，165个基因一致地甲基化。甲基化基因在所有染色体上呈均匀分布。这些的基因本体也表明甲基化的基因之间没有功能相关性。我们在IGF2中检测到甲基化的存在，IGF2是一种印记基因，因此已知含有DNA甲基化。该方法是非常具体的，并提供了一种快速和有效的方法来分析甲基化景观的高通量规模。该方法使用现有技术来评估甲基化，因此可以非常有效地整合到所使用的任何平台中。
In this work we demonstrate a novel method of methylation detection that utilises immunoaffinity to detect the presence of methylated DNA hybridised to a cDNA microarray. We use a monoclonal antibody specific to 5 methyl cytidine to detect the presence of 5 methyl cytosine in genomic DNA from human fibroblasts bearing the karyotype 45 XO. We report that over 2900 genes show the presence of methylation in this condition. We also report that 165 genes are consistently methylated in all replicates of these experiments. The methylated genes show a uniform distribution over all the chromosomes. The gene ontology of these also indicates no functional correlation between the genes that are methylated. We detect the presence of methylation in IGF2, an imprinted gene and thus known to harbour DNA methylation. The method is extremely specific and offers a quick and efficient way to analyse the methylation landscape on a high throughput scale. This method uses existing technology to assess methylation and thus can integrate very efficiently into any platform used.