vector for stem cell differentiation - (Sep/07/2006 )
my immediate task is to overexpress a certain gene in human ES cells and the over expression should continue even when it is differentiated into a neuron. my vector has a CMV promoter which functions only in undifferentiated state. are you aware of any vector that contains a promoter that could function even in differentiated state.
This is an excellent question that I've run into myself, albeit not with ES cells but within cell lines. I haven't found a functional constitutive promoter within differentiated cells, and have instead worked around this by transfecting my cells ~24h prior to differentiation to allow for the product of interest to be expressed. Alternatively, you can try a promoter from a gene you're sure is highly expressed in differentiated cells.
CMV is an RNA plymerase II promoter. I've had good results using U6/H1 RNApolIII promoters in differentiated cells but this is not generally the type you'd want for cDNA overexpression. Hopefully someone can suggest an alternative.
Suggestion try with PGK phospho glycerate kinase promoter all PGK vectors (constitutive expression) -