5-Hydroxymethylcytosine – To switch a gene on or off
The widespread adoption of emerging epigenetics technologies has provided the scientific community with new tools for understanding genetic inheritance phenomena. These advanced tools include kits and equipment for DNA or chromatin shearing, chromatin immunoprecipitation (ChIP, ChIP-Seq), methylated DNA immunoprecipitation (MeDIP), methyl binding domain (MethylCap, MBD) studies, bisulfite conversion, automation (IP-Star, SX8G Compact), and Next Generation sample preparation.
In recent years, interest in DNA methylation modifications has increased, as abnormal DNA methylation status is often correlated with disease. One particular DNA modification, 5-methylcytosine or 5-Methylcytidine (5-mC), can switch a gene “on or off,” and is the most extensively studied modification among the known plant and vertebrate epigenetic marks. The 5-mC modification is also implicated in genomic imprinting and contributes to the of control gene expression. This modification occurs when DNA methyltransferases (DNMTs) catalyze the reaction of a methyl group to the fifth carbon of cytosine in a CpG dinucleotide.
Researchers recently discovered another important DNA methylation modification, the 5-hydroxymethylcytosine (5-hydroxy methylcytosine, 5-hmC) which occurs after the enzymatic conversion of 5-methylcytosine into 5-hydroxymethylcytosine by the TET family of cytosine oxygenases. The5-hydroxymethylcytosinecould be involved in a pathway to demethylate DNA, as seen by the evidence that 5-hydroxymethylcytosine is repaired as mismatched DNA and replaced with unmethylated cytosine.
Diagenode, the pioneer in the field of epigenetics research, facilitates DNA methylation studies with optimized assay protocols and robust products. The company provides kits, reagents, antibodies, equipment, and protocols dedicated to DNA methylation studies at the single loci or genome-wide level. The antibodies product line includes 5-hmC, 5-mC, and a Cytosine DNA Standard Pack which includes hydroxymethylated, methylated, and unmethylated DNA standards. These standards have been produced by PCR using the Diagenode’s MethylTaq DNA polymerase or5-hm dCTP (for the hydroxymethylated standard). Each standard either contains normal cytosines, 5-methylcytosines, or 5-hydroxymethylcytosines. All protocols and products are available for manual assays or for automated assays with the SX-8G IP-Star Compact automation workstation.
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