Adenoviral vectors

For the generation of adenoviruses we are currently using kits from Clontech. The viral vector is generated by enzyme mediated insertion directly from a purified PCR product of the transgene into the the E1 and E3 deleted Ad5 genome. Transgene expression is driven by the strong CMV promoter.

Viral production is performed by transfection of the linearized Ad-vector into HEK293 cells expressing E1 and E3 genes of the Ad5 genome and several rounds of amplification. Alternatively, existing adenoviruses can simply be used to start the amplification. Purification of properly assembled adenoviral particles is done by caesium chloride density gradient followed by dialysis.

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