Transfection of RNA molecules transcribed from T7D-polio with an authentic 3' end into mammalian cells produced wild-type poliovirus with an efficiency similar to that of virion RNA.
In vitro synthesis of poliovirus RNA with an authentic 3' polyadenine sequence yields infectivity comparable to native virion RNA.
It has been shown by van der Werf et al. (S. van der Werf, J. Bradley, E. Wimmer, F. W. Studier, and J. Dunn, Proc. Natl. Acad. Sci. USA 83:2330-2334, 1986) that in vitro synthesis of poliovirus RNA by T7 RNA polymerase gives rise to infectious RNA molecules; however, these molecules are only 5% as infectious as RNA isolated from virions. A plasmid, T7D-polio, was constructed that allows the in vitro synthesis of full-length RNA molecules with two additional guanine residues at the 5' end. However, T7D-polio differed from the construct of van der Werf et al. in that RNA transcribed from T7D-polio has an authentic 3' end, ending with only a polyadenine nucleotide sequence. Transfection of these RNA molecules into mammalian cells produced wild-type poliovirus with an efficiency similar to that of virion RNA. The use of this vector in the characterization of viral mutants in vivo and in vitro is discussed.
Peter Sarnow (Sun,) conducted a other in Poliovirus infection (in vitro). T7D-polio plasmid transcripts (RNA with authentic 3' end) vs. Virion RNA was evaluated on Infectivity / production of wild-type poliovirus. Transfection of RNA molecules transcribed from T7D-polio with an authentic 3' end into mammalian cells produced wild-type poliovirus with an efficiency similar to that of virion RNA.