Research experts in the epigenetic field from Epigentek have developed a new, breakthrough approach for the identification of the "sixth DNA base," 5-hydroxymethylcytosine (5-hmC). This technology is based on a high throughput, strip-well format and is also incorporated into the first commercially available product, the MethylFlash(TM) Hydroxymethylated DNA Quantification Kit (http://www.epigentek.com/hydroxymethylation), for rapidly quantifying hydroxymethylated DNA.
5-hmC is a modified form of 5-cytosine, recently discovered in animal tissues. The function of 5-hmC in epigenetics may be different from its forerunner 5-methylcytosine (5-mC) and currently remains a mystery. It is believed, though, that 5-hmC plays an important role in switching genes on and off. The presence of 5-hmC makes it necessary to not only re-evaluate existing DNA methylation data, but also necessary to determine relative distribution and changes of 5-hmC in human tissues of healthy and diseased statuses. Prior to Epigentek's MethylFlash(TM) technology, there were no methods that could be used for practically or routinely identifying 5-hmC and discriminating this base from 5-mC.
Epigentek's new method can be used for rapidly and accurately identifying hydroxymethylated DNA or 5-hmC in a high throughput format, suitable from any species including mammals, plants, fungi, bacteria, and viruses in a variety of forms including cultured cells, fresh and frozen tissues, paraffin-embedded tissues, plasma/serum samples, and body fluid samples. In the assay, DNA is bound to strip wells that are specifically treated to have a high DNA affinity. The hydroxymethylated fraction in the DNA is then immunospecifically detected.
Utilizing this method, scientists at Epigentek for the first time in the world identified that 5-hmC is abundant in normal human brains and colon tissues but significantly decreased in colon cancer tissue/cells. It was also found that 5-hmC is more than 30% and 15% as abundant as 5-mC in human brains and colon tissues, respectively. This discovery demonstrates that this new method would be powerful tool in further unraveling the mystery of epigenetics.