Hello Pranav.
Yes, you can if the virus is prepackaged with pCMV-GFP. This will show GFP expression in any cell lines. However, if it has certain cell-specific promoter (i.e., cardiac specific ), you can still transduce with no expression. Make sure that the promoter is either CMV or beta-actin promoters. Depending on your virus used for packaging GFP, expand virus particles and store them until use for transduction according to reliable methods.
Reagrds.
Dear Sang Ho Lee, the viral vector has GFP as the expressing transgene and not a tag which takes time to express. The mammalian cell ine on the other hand has its own cycle. How do I coordinate these two. Could you suggest if it is now possible.
Hello Pranav.
When I wrote pCMV-GFP, it did not mean a tagging some molecule at all. What it means is that viral vector has GFP gene. And when it get infection and get inserted into the genome of your eukaryotic cells, by the action of CMV promoter is activated by transcrition factors present within the eukaryotic cells. So it is not a tagging, it is a promoter driving GFP gene expression.
By the way, does your virus then have its own promoter? If so, it will take a prolonged time for unoptimized packaging until it realized as a serotype expression depending on cell phenotypes.
My advice is to provide more detail of your work, then, I am sure, some virologist would give immediate answers or current problems or cell types you should use.
Best wishes.
Dear Dr. Lee ,
My vector has its own promoter CAG (Chicken -beta-actin hybrid)
It is AAV-DJ8 (Capside from AAV-DJ and ITR from serotype 2) with a transgene for eGFP.
The problem is simply as you mentioned that it takes a prolonged time till it starts expressing its gene.
I need to try and see how efficiently the transgene is expressed (eGFP) in -vitro but because the cell line . HEK293 that I use has its own time period of growth, I am not sure ho to proceed.
It would be really helpful if some experts can assist me a bit.
Regards
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