I am using a commercially available viral vector AAV-DJ (Capsid from AAV-DJ and ITR from serotype 2) with a transgene for eGFP.
My vector has its own promoter CAG (Chicken -beta-actin hybrid)
The problem is simply that the viral vector takes a prolonged time till it starts expressing its gene (eGFP).
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 you can assist me a bit.
Regards
Could you please define "prolonged time"? It shouldn't take more than 48h - if it is something like 5-7 days, have you checked if it really is GFP fluorescence or autofluorescence from dead/dying cells?
I haven't tried it yet but am planning it really soon.
In general , I want to try out the if the available serotype expresses efficiently in-vitro in my HEK293 cell culture.
What I am concerned about is that the expression time of the viral serotype might take longer than the 48 hr time period that my cells take from seeding to the next split.
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