Research activity
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2) Functional characterization of 3'UTRs of specific genes target for miRNAs |
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Following previous studies, we predicted polymorphic 3'UTRs where a single nucleotide polymorphisms (SNPs) were, in silico, able to affect the binding with specific micro-RNAs. The SNPs were assayed in a case-control study and we identified two of them (within CD86 and INSR) associated with the risk of colorectal cancer. In order to affirm that the risk is causally related to the deregulation of these genes, via a mechanism involving the post-transcriptional regulation by miRNAs, we are verifying whether the predicted 3'UTRs are truly target sites for the predicted miRNAs and whether the SNPs affect the strength of the binding. |
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The experiments are carried out on cell lines transfected with the Renilla/Luciferase gene reporters, where the 3'UTRs of the Luciferase is replaced by the 3'UTRs of the two genes (either in their common and variant forms). Synthetic miRNAs are also transfected in order to measure the Luciferase/Renilla ratio under different miRNA binding, to detect whether the polymorphism truly affect the binding to the 3'UTR. |
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The preliminary studies are reported here. |
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