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CMV promoter driven transgene expression


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4 replies to this topic

#1 moljul

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Posted 09 June 2009 - 12:06 AM

hello all,

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

#2 genehunter

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Posted 09 June 2009 - 08:51 AM

Maybe a little more detail on how you did the transfection would help us to help you on this.

#3 moljul

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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.



110 DNA komplexed with PEI (N/P ratio 3,3). injection i.v. via tailvein.

#4 Rsm

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Posted 10 June 2009 - 05:06 PM

Which cells did you check? The CMV promoter does not work in some cells...
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#5 moljul

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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!




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