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Updated: Jun 28, 2026

Applying an Inducible Expression System to Study Interference of Bacterial Virulence Factors with Intracellular Signaling
Published on: June 25, 2015
Multiple shRNAs expressed by an inducible pol II promoter can knock down the expression of multiple target genes
Xu-Gang Xia1, Hongxia Zhou, Zuoshang Xu
1Department of Biochemistry and Molecular Pharmacology, University of Massachusetts Medical School, Worcester, MA 01605, USA. Xugang.xia@umassmed.edu
Abstract:
RNA interference (RNAi) has been increasingly used for reverse genetics. Both pol III and pol II promoters have been used to synthesize short hairpin RNA (shRNA) for knockdown of gene expression in mammalian cells and animals. Compared with gene knockout approaches, RNAi has the advantage of being simple, quick, and low cost. Here we describe a method that enhances this advantage where knockdown of expression of multiple genes in the same cells is required. A tetracycline-regulated pol II promoter construct allows the expression of up to three shRNA genes that have been cloned into introns of a transcript bearing green fluorescent protein (GFP) coding sequences. This method may be used to establish stable knockdown cell lines and may also prove useful for investigating gene-gene interactions in transgenic animals.
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