A novel method to assess the full genome methylation profile using monoclonal antibody combined with the high throughput based microarray approach.
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 2,900 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.
Kelkar
Ashwin
A
Senior Research Fellow, Molecular Biology Research Laboratory, University of Pune, Pune, India.
Deobagkar
Deepti
D
D000911:Antibodies, Monoclonal; D003562:Cytidine / Q000031:analogs & derivatives / Q000378:metabolism; D019175:DNA Methylation; D023281:Genomics / Q000379:methods*; D006801:Humans; D009693:Nucleic Acid Hybridization; D020411:Oligonucleotide Array Sequence Analysis / Q000379:methods
D000911:Antibodies, Monoclonal; D003562:Cytidine; C016568:5-methylcytidine