I am currently trying to clone a particular cytokine gene's regulation - final product would be a 5' promoter element, GFP (in place of the original gene's coding region), and 3' UTR, into a lentiviral plasmid to label tumor cell populations.
I'm curious if the GFP signal would be an accurate proxy for levels of expression of the gene of interest.
For that reason, I'm concerned about the effects of WPRE on the construct. I understand that WPRE's are good for expression efficiency, but I'm fuzzy on the mechanism. Does WPRE affect mRNA stability?
I also wanted to do transcription factor studies using the above construct. Should I move that expression casette into a non-lentiviral plasmid before continuing?