Key result
Transfection of naked influenza virus-like RNAs into cells co-expressing polymerase subunits and nucleoprotein from SV40 recombinants was as effective in directing protein synthesis as standard transfection.
Naked influenza virus RNA can be expressed upon transfection into cells co-expressing the three polymerase subunits and nucleoprotein, potentially allowing rescue of influenza viruses without a helper virus.
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SV40-expressed influenza polymerase and NP support viral RNA expression in vivo; leaves open broader reverse genetics applications.
The functionality of the influenza virus polymerase subunits and the nucleoprotein expressed from simian virus 40 (SV40) recombinants has been tested by their ability to direct the in vivo expression of influenza virus-like RNAs. These RNAs, which contained either the chloramphenicol acetyltransferase (CAT) or haemagglutinin (HA) genes, were synthesized and reconstituted in vitro into viral ribonucleoproteins with a polymerase/nucleoprotein mixture purified from influenza virus-infected cells. Only the coinfection with SV40 recombinant viruses expressing the three polymerase subunits and the nucleoprotein allowed the expression of the transfecting CAT or HA RNAs, confirming that this set of viral genes is the minimal requirement for viral gene expression. Unexpectedly, transfection of the corresponding naked RNAs into SV40 recombinant-infected cells was as effective in directing the synthesis of CAT or HA proteins as the standard reconstituted ribonucleoprotein transfection. These results may be important for the genetic analysis of trans-acting factors involved in influenza virus transcription and replication and may open the way to rescuing influenza viruses in the absence of a helper virus.
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Luna et al. (1993) studied Influenza virus infection model. Transfection of naked influenza virus-like RNAs vs. Standard reconstituted ribonucleoprotein transfection was evaluated on Expression of transfecting CAT or HA RNAs. Transfection of naked influenza virus-like RNAs into cells co-expressing polymerase subunits and nucleoprotein from SV40 recombinants was as effective in directing protein synthesis as standard transfection.
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