Description
Co-transfection of mammalian cells with the pAdVAntage™ Vector enhances transient protein expression in a variety of cell types by increasing translation initiation. Luciferase constructs co-transfected with the pAdVAntage™ Vector showed at least a tenfold increase in luciferase expression in 293 and HeLa cell lines over transfections performed with the construct DNA alone.
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's antiviral defense system. DAI phosphorylates the translation initiation factor eIF-2, halting translation and therefore protein production.
However, DAI translation inhibition can be overcome with the adenoviral Virus Associated I RNA (VAI RNA) 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.
References
Farrell, P.J. et al. (1977) Cell 11 , 187–200.
Groskreutz, D. and Schenborn, E. (1994) Promega Notes 48 , 8–12.
Kitajewski, J. et al. (1986) Cell 45 , 195–200.
O'Malley, R.P. et al. (1986) Cell 44 , 391–400.
Sequence Information
pAdVAntage™ Vector GenBank® Accession Number U47294 and vector sequence text file .
