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Delivering siRNA Using Adenoviral Vectors

of GFP with pSilencer adeno. HeLa cells were infected (MOI = 160) with recombinant adenovirus derived from pSilencer adeno 1.0-CMV engineered to express an siRNA targeting GFP. Cells were analyzed 48 hours after infection by fluorescence microscopy.


How to Use pSilencer adeno 1.0-CMV
First an oligonucleotide insert encoding the desired siRNA is cloned into the pre-linearized shuttle vector (provided), which includes a modified CMV promoter for efficient expression of the siRNA sequence. The shuttle vector containing the desired sequence is then linearized and introduced into HEK 293 packaging cells (not included) along with the linearized adenovirus backbone (Figure 3). Inside the HEK 293 cells, the shuttle vector and the adenoviral backbone plasmid recombine to produce an adenoviral genome including the siRNA expression cassette. The adenoviral genome is packaged into a protective coat and released into the culture medium. These adenoviral particles are then harvested and used to infect cells or animals. Adenoviral particles can be stored for long periods of time, so experiments can be performed long after adenovirus production.

Figure 3. Schematic of pSilencer adeno Recombination and Packaging in HEK 293 Cells. The shuttle vector and backbone plasmid are linearized and then transfected into HEK 293 cells. Inside the cell, the plasmids recombine and the recombined genome is replicated and packaged into adenoviral particles. mu=map units.


The Complete Adenoviral Production System
The pSilencer adeno 1.0-CMV System includes everything needed to produce f
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