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miRNA overexpression for target validation - (Sep/15/2010 )

I want to experimentally show that if a given mRNA is a true target of a specific mRNA, then modulation of miRNA concentration should correspond to a predictable change in the protein encoded by the target mRNA AND that If a given miRNA is involved in the regulation of a target gene, then this regulation equates to changes in biological function. Both are part of a project that's validating a target(influenza virus binding protein) for 4 miRNAs. How can I do this in as simple and unexpensive (as possible). Yes, I'll take any ideas. Thank you all


For miRNA targeting, because the miRNA-target relationship is not one-to-one, but one-to-many, it is hard to use concentration-function correlation to validate a true target. The standard method of doing this is to use a reporter with the target or mutated target cloned into the 3UTR of the reporter gene. If the muation disrupts inhibition of the reporter gene by the miRNA, it is suggested that the target is a true one.