I'm looking for a methode to demethyle DNA in vivo, during more than 5 days. All I've read about 5-aza-CdT is for maximum 5 days. Why ?
Is there another drug and if yes, how does it work ?
thanks
Elea -Elea-
Hi Elea,
Max. 5 days, that's not right. It all depdents on the sensitivity of your cells to the drug. We have treated cells continously for two weeks.
Hope that helps. -pcrman-
Elea,
it also depends on the cell line or cell type you are using, 5-azadC has a rather short half life and can be metabolised by nucleoside deaminase. If your cell is expressing deaminase then it is resistant to this drug.
The drug works as a nucleotide analog to Cytosine that can covalently bind to DNA methyltransferases and therefore prevents the ability for the enzyme to methylate DNA.
This drug must be incorporated into the DNA (ie: your cells must be replicating) for it to work.
Zebularine (1-(beta-D-ribofuranosyl)-1,2-dihydropyrimidin-2-one) is another nucleotide analogue that has been shown to last longer than 5-aza-dC (see Cheng et al 2004 Mol Biol Cell) and is not affected by deaminase activity.
Zebularine is also available commercially.
I am currently treating CHO hybrids with 5-aza-dC it's fun fun fun!! -methylnick-
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