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pSilencer 4.1-CMV: Versatile Vectors for Expression of siRNA, miRNA, and mRNA


Ambion's pSilencer 4.1-CMV plasmid vectors are designed for long-term gene silencing experiments in a broad range of cell lines. The vectors carry an RNA polymerase II-type CMV promoter (human cytomegalovirus immediate-early promoter) and an optimized SV40 polyadenylation signal to drive high-level expression of hairpin siRNA. For stable transfections requiring long-term antibiotic selection, an SV40 promoter expresses one of three antibiotic resistance genes (hygromycin, neomycin, or puromycin) for long-term antibiotic selection. Here, we show that pSilencer 4.1-CMV is a versatile vector that can be used to express siRNA as well as microRNA (miRNA) or messenger RNA (mRNA).


Efficient Gene Silencing

To demonstrate the utility of the pSilencer CMV vectors in RNA interference experiments, a pSilencer 4.1-CMV vector was engineered to express a hairpin siRNA targeting GAPDH. This construct was then transfected into HeLa cells. As shown in Figure 1, the level of GAPDH protein was reduced by ~80% two days after transfection. This suggests that the hairpin siRNA was efficiently expressed and correctly processed in the cell to generate a mature, functional siRNA. Long-term antibiotic selection of the transfected cells produced cell lines exhibiting reduced levels of GAPDH mRNA and protein for over 4 months (see

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