We integrated a gene in a cell line genome, when we culture them, the gene turn to silencing. We collected non-silencing cells and culture them, some of them turn to silencing again. We already check the gene, it is fine and still there.
We want to figure out what factors cause the silencing, but we are not familiar to epigenetic.
Any one and any suggestions will appreciate.
A more common cause is that the gene product is in some way toxic or deleterious to your cells. Since you did not provide more details, I cannot be more specific. When this is the case, you are effectively selecting against high expressors. We had this problem when working with plant protoplasts and electroporation of genes involved in anthocyanin production. The solution was to use an inducible promotor, which prevented expression of the introduced genes just after the electroporation. Only after treatment with the inducing agent cells would be triggered to express the introduced genes, at the time of our bidding.
I hope this is helpful. Kind regards, Harrie Verhoeven
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