Hello everybody,
I have tried to express my GOI in RPE1 cells under the control of a hEF1a promoter. While such construct is weakly expressed, GOIs under the CMV promoter appear to work just fine.
I have made some research but apparently there is no study about exogenous promoters activity in this cell line. Have you ever experienced this?
Thanks in advance
Francesco
Hi,
unfortunately I do not have any information on this particular promoter/cell line combination, however generally EF1a is considered weaker than most viral promoters (SV40, CMV, ...), but more consistent in expression strength. CMV can and will be modulated by many factors (stress, sodium butyrate, and likely many more), however EF1a tends to remain quite stable. Likewise, there are cell lines where CMV does not work at all (several nonspecific antiviral mechanisms, e.g. silencing by methylation).
My advice is, when you want to get a gene expressed at a stable level (e.g. for normalization purposes) use EF1a, if you just want to highly express a gene and dont care about the exact amount or variability (e.g. for protein expression), take CMV.
Thank you Patrick,
I agree with you, that's why I have always used the EF1a for any cell line I worked with. I was just puzzled to see that this promoter seems not to be as effective for RPE1 cells.
Thanks
EF1 is very good in neuronal cells so what you need to do use CAG promoter or hSyn (strongest promoter i ever seen).
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