siRNA to shRNA...trouble! - (Sep/28/2010 )

Hello everybody!
I'm trying to down-regulate a protein... I've found a sequence that work perfectly as siRNA but when I "transformed" it into shRNA (and put it in the pLKO-Tet-On vector) it doesn't work at all! I've followed completly the datasheet instruction to make the loop, so I guess that this is not the problem...

Someone can help me?!
My boss is making me going mad!

The successful rate of converting siRNA to shRNA is about 70%.
A lot of siRNA design was not compatible with Zamore rules, so I will suggest you to go to different online shRNA designer to see whether you can get one close to your siRNA sequence.