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Has anyone seen target mRNA increasing after RNAi? - (Apr/11/2006 )

I am using dsRNA in Drosophila S2 cells to do RNAi knockdown. For tow genes, I detected increased mRNA using RT-PCR. I used full-length cDNA of the two gene to generate dsRNA. I wonder if anyone has seen this happening? BTW,four other similar experiments resulted in decreased mRNA.
Thanks

-leemovie-

At least two people in this forum have mentioned that they got enhanced expression of siRNA target genes. Although the situation may differ, but some possibilities are:

1) an interferon response triggered by siRNA or shRNA
2) siRNA mistargets another gene that in turn activate the true target gene
3) squence-specific gene activation by unknown mechanisms (this is possible!!!)

-pcrman-

We have such experience in RNAi for plants and also learn a case from a friend, he said it might be many people found such change but nobody said that in paper yet...
by the way, you mentioned that you are using full-length cDNA of the two gene to generate dsRNA, although I am not sure the detail, but is there any chance that the transcript of the full-length cDNA has been detected by RT-PCR?

-rye-

QUOTE (leemovie @ Apr 11 2006, 08:58 AM)
I am using dsRNA in Drosophila S2 cells to do RNAi knockdown. For tow genes, I detected increased mRNA using RT-PCR. I used full-length cDNA of the two gene to generate dsRNA. I wonder if anyone has seen this happening? BTW,four other similar experiments resulted in decreased mRNA.
Thanks

sad.gif I have encountered the same problem. Can anyone give me a possible explaination?

-jelly-

I also had this problem last year. I found a very large mRNA expression enhancement, but thought it was a mistake. Just used a different siRNA sequence ...

I am glad someone else had the same problem. Just wondering what is it about. Could be a not known process?

-Bruja-

Maybe the target gene has a negative feedback loop which is interrupted with the siRNA, so the gene finally is overexpressed ...

-Bruja-

try this citation

Small dsRNAs induce transcriptional activation in human cells Hideki Enokida, and Rajvir Dahiya

Long-Cheng Li, Steven T. Okino, Hong Zhao, Deepa Pookot, Robert F. Place, Shinji Urakami,

PNAS published online Nov 3, 2006;

-94121-

siRNA enahnced transcription in a sequence specific way. Interesting!

-genehunter-1-