Hi, community!
As part of my PhD project I have been trying to transduce mouse HSCs (Lin- or 5-FU treated Bone Marrow cells) with a lentivirus for subsequent transplantation and in vivo expression of my GOI.
The human PGK promoter i used initially to drive expression in mouse HSCs appeared to work well in vitro. In vivo, however, I could detect very little expression. My reporter gene driven by SFFV seems to be expressed just fine, leading me to believe that poor GOI expression is a promoter issue.
Faced with the task of redesigning my expression vector, I have two alternative promoters readily available - human CMV and human Ubiquitin. Hence my question is, are either of those expressed well in murine cells and which would be the preferred one? Should I be using human promoters to drive expression in mouse in the first place or are murine promoters always the better option?
Any advice is greatly appreciated,
Many thanks!
My suggestion would be to try EF-1 alpha promoter.
EF-1 alpha is often useful in conditions where other promoters (such as CMV) have diminished activity or have been silenced (as in embryonic stem cells).
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