2D protein gel for miRNA target screening? - (Jan/23/2007 )
Is 2D sensitive enough to find the targets of a miRNA? Those miRNA targets may not be abundantly expressed. So how to remove those abundant proteins, like tubulin, lamin, etc.,? Thank you!
From what we know so far, it seems that microRNA regulates more genes by affecting mRNA levels than inhibiting translation. So cDNA expression profiling probably is the right apporach and also easier to do. How does microRNA affect mRNA? there is no clear answer yet. Possible mechanisms include microRNA affects mRNA decay in P bodies.
Thank you pcrman.
To my knowledge, when miRNA perfect match with it's target mRNA, the target can be knockdown by mRNA degradation. If not perfect match, the RNAi machinery will not be activated, instead, the target mRNA is knockdown through translational repression.
If you are right, so I can over-express a miRNA in non-expressing cell, then use cDNA microarray to find the affected genes.