In this study, we designed a short hairpin RNA vector-based oligodeoxynucleotide (VB-ODN) carrying transcription factor (TF) consensus sequence which could function as a decoy to block TF activity. Specifically, VB-ODN for Nuclear factor-kappa B (NF-kappa B) could inhibit cell viability and decrease downstream gene expression in HEK293 cells without affecting expression of NF-kappa B itself. The specific binding between VB-ODN produced double-stranded RNA and NF-kappa B was evidenced by electrophoretic mobility shift assay. Moreover, similar VB-ODNs designed for three other TFs also inhibit their downstream gene expression but not that of themselves. Our study provides a new design of decoy for blocking TF activity. (C) 2014 Elsevier Inc. All rights reserved.