pAdVAntage™ Vector Technical Bulletin

Literature # TB207

Cotransfection of mammalian cells with the pAdVAntage™ Vector enhances transient protein expression in a variety of cell types by increasing translation initiation. Transfection of mammalian cells with an expression vector often results in suboptimal expression of the protein of interest. Double-stranded RNA (dsRNA)generated during transfection is thought to activate the dsRNA-activated inhibitor (DAI), one of several enzymes involved in the host cell antiviral defense system. DAI phosphorylates the translation initiation factor eIF-2, halting translation and therefore protein production.

However, inhibition of translation by DAI can be overcome with the adenoviral Virus Associated I RNA (VAI RNA) that is produced by RNA polymerase III following co-transfection with the pAdVAntage™ Vector. The VAI RNA binds to DAI, preventing its activation, thereby allowing translation and protein expression. The pAdVAntage™ Vector contains base pairs 9,831–11,555 of the adenovirus type 2 genome on a 1,724bp SalI-HindIII fragment that encodes the virus-associated RNA genes, VAI and VAII.