CMV promoter driven transgene expression
Posted 09 June 2009 - 12:06 AM
I did some in vivo transfection experiments using pcDNA3.1 vectors. The transfection efficiency I checked with PCR! after 36h the expression decreased to baseline. that is too early I think. Could there be some kind of repression or other cellular processes which decrease transcriptional activity?
thanks for suggestions
Posted 09 June 2009 - 08:51 AM
Posted 10 June 2009 - 11:36 AM
Maybe a little more detail on how you did the transfection would help us to help you on this.
110µ DNA komplexed with PEI (N/P ratio 3,3). injection i.v. via tailvein.
Posted 10 June 2009 - 05:06 PM
Posted 11 June 2009 - 01:36 AM
Which cells did you check? The CMV promoter does not work in some cells...
i check no specific cell type. its the kidney where my transgene should be expressed. i have expression for 24-36h, this i checked by western blot. but whats after this time? i know that there is a lot of vector DNA in it!