I generated stable cell lines expressing a mouse protein using a lentivirus expression system. I have used different parent cells, Vero cells (monkey cell line) and LA-4 cells (mouse cell line). In both cell lines, lentivirus integration was good and further enriched after sorting for an MCherry reporter protein. I used Flag tag as the reporter of my protein of interest. My protein was expressed in LA-4 cells (> 80% cells flag positive). However, the expression of this protein is very low and not reproducible in Vero cells. Does anyone have any idea why this expression is low? And how can I improve the expression of this protein in Vero cells?
It seems to me like your answer is in your clues. You are trying to express a mouse protein, and your mouse cell lines will have the most ideal environment of chaperone, splicing factors, transcription and translation factors, etc, for that mouse gene. There are subtleties to the intra-cellular environment that are preventing the mouse protein from being well expressed in a monkey cell line.
Dear Melkamu,
in my opinion , chaperones, splicing factors, transcription and translation factors comparable in both species and should be adequalely active in both mammalia cell types. However, in general cell lines differ in their capacity to be transfected, to respond to viral DNA integration, and to express proteins (esp. from other species).
In my opinion, you should revise your protocol as well as the experimental setting. Why are you using this specific monkey cell line? Is there any alternative cell line better described regardig transfection via lentiviral system or expression of murine proteins? Are concentation of viral units optimized for each of both species? etc.
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