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Superior Gene Silencing Using Adenoviral Vectors

s virus particles that can be harvested for experiments. Harvested adenoviral particles can then be used to infect cells or animals, or stored for future experiments.

Figure 1. Schematic of pSilencer Adeno 1.0 CMV System Recombination and Packaging. The shuttle vector and backbone plasmid are linearized and then transfected into HEK 293 cells. The HEK 293 packaging cells contain the EIA and EIB genes which are not present in the introduced vectors and are required to form infectious particles. Inside the cell, the plasmids recombine forming a functional genome that is replicated and packaged into adenoviral particles.


siRNA from Adenoviral Vectors Reduce Expression from Transgenic Alleles in vivo
To demonstrate that adenoviruses are useful for decreasing gene expression via the RNAi pathway in animals, Dr. Beverly Davidson and colleagues, our collaborators at the University of Iowa, injected recombinant adenoviruses expressing siRNAs targeting green fluorescent protein (GFP) or -glucuronidase (a negative control) into the brains of transgenic GFP-expressing mice (1). Five days after injection, brain sections were analyzed by fluorescence microscopy and Western blot (Figure 2). GFP expression was reduced in the injected brain hemisphere, indicating the utility of the method for reducing gene expression in animals.

Figure 2. Silencing of Transgenic GFP in Mouse Brain with an Adenoviral Vector (1). Adenovirus engineered to express an siRNA targeting GFP (A, top panels) or a negative control siRNA targeting human -glucuronidase (A, bottom panels) were injected into t
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