A gene expression carrier for regulating NF-κB activity in a cell and a regulation method therefor and an application thereof, the carrier comprising two sequence elements, a promoter sequence that regulates gene expression and a promoter downstream miRNA coding sequence; the promoter sequence consists of an NF-κB decoy sequence and a minimum promoter sequence; the miRNA coding sequence is a coding sequence capable of encoding miRNA of target NF-κB mRNA. The transfection of the gene expression carrier may moderately and effectively reduce NF-κB activity within a cell having over-active NF-κB activity and has no obvious effects on the activity of normal cells, thereby preventing side effects thereof. The regulation method for a gene expression carrier is simple, fast and effective. The gene expression carrier is used for preparing an NF-κB activity regulation reagent or drug molecule for the gene therapy of diseases closely related to NF-κB over-activity, being a novel gene therapy reagent.
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