Single nucleotide polymorphism associated with mature miR-196a2 influences the expression of Lymphocyte-specific protein 1(LSP1) gene

Objective:To investigate the effect of a common polymorphism T/C (rs11614913) in mature miR-196a2 on the expression of LSP1 gene. Method:The pCDNA3.1-miR196a2-C and pCDNA3.1-miR196a2-T expression plasmids were created containing T or C allele of pre-miR196a2. Meanwhile,mature miR-196a2-C and miR-196a2-U probes were chemically synthesized for the investigation. The 3’UTR from LSP1 gene was cloned downstream of the firefly luciferase gene in pMIR-REPORTTM Luciferase Vector. Two sets of cotransfection were performed with CHO,HEK293 and A549 cells,and the luciferase activity with the Dual-Luciferase Reporter Assay System were analyzed. Results:pCDNA3.1-miR196a2-C construct group showed significantly lower levels of luciferase expression compared with pCDNA3.1-miR196a2-T construct group(P 0.05),when LSP1 3’UTR luciferase reporter plasmids were cotransfected with miR-196a2 expression plasmids in CHO,HEK293 and A549 cells. Mature miR-196a2-C probe group showed significantly lower luciferase activity compared with miR-196a2-T probe group (P 0.05),while LSP1 3’UTR luciferase reporter plasmids were cotransfected with mature miR-196a2 probes in CHO,A549 and HEK293 cells. Conclusion: The miR-196a2 could effectively bind the predicted target gene of LSP1 and influence the expression on the level of transcription,while miR-196a2 carrying the rs11614913 C allele.