problem with miRNA functional study - (Jul/28/2011 )
Hey guys, I got headache problem with my miRNA functional study. I transfected with the miRNA, and then did a lot of western blot. and then i found all the proteins I tested were up-regulated by the miRNA, including those apparently act opposite roles (e.g. pro-apoptosis vs. anti-apoptosis; cell cycle arrest vs. proliferation related protein) and those predicted as target proteins! even more surprisingly, there was one predicted target which showed promising results in the 3'UTR-luciferase assay, again appeared up-regulated in the western blot. I am sure I applied the samples in the right order in all the assay. Anyone advise why it is happening?! Thanks!
It has been reported mainly by Joan A. Steitz group that certain miRNAs can activation protein translation by targeting 3UTR region when cells are under stressed condition. What you saw could be real.