How to explain this luciferase result?!
Posted 04 June 2011 - 12:49 AM
I have performed dual luciferase reporter asay to check if my microRNA binds to the potential target gene. I co-transfected the luciferase reporter wit the 3UTR (psicheck) and my microRNA, but i didnt observe any changes in the renilla/firefly ratio, compared to cotransfection with the vector control of the microRNA. However, when i co-transfected microRNA with the mutated 3'UTR luciferase vector, the ratio increased, compared to the empty vector of miRNA as well as the wild-type form. How to explain this weird result?
a) wild-type luciferase reporter + empty vector : 1
wild-type luciferase reporter +microRNA: 1
c) mutant luciferase reporter +empty vector: 1
d) mutant luciferase reporter + microRNA: 1.3
Posted 05 June 2011 - 08:57 AM
Posted 23 July 2011 - 08:39 PM
there are two miRNA binding sites predicted to target at the gene, and mutation seed region of each sites can relieve the repression of miRNA to 3-5 fold of that in wildtype. However, the unexpected result is that double mutation strengthen the repression instead of removing the repression by this miRNA. What's wrong with it?
Any suggestion are appreciated!
Edited by whitedolphin, 23 July 2011 - 08:59 PM.