Construct a reporter vector containing the candidate promoter fragment, and analyze promoter activity strength with a dual-luciferase reporter assay.
Use EMSA to validate the direct binding relationship between the candidate transcription factor and the promoter cis-element.
Construct a reporter vector containing the 3'UTR fragment, and use a dual-luciferase assay to validate the candidate miRNA's targeted repression of the target gene's 3'UTR.
Combine the above methods to resolve the regulatory relationships within the miRNA-mRNA-lncRNA competing endogenous RNA network.
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