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An improved reverse genetics system for influenza A virus generation and its implications for vaccine production.
- Source :
-
Proceedings of the National Academy of Sciences of the United States of America . 11/15/2005, Vol. 102 Issue 46, p16825-16829. 5p. - Publication Year :
- 2005
-
Abstract
- The generation of vaccines for highly pathogenic avian influenza viruses, including those of the H5N1 subtype, relies on reverse genetics, which allows the production of influenza viruses from cloned cDNA. In the future, reverse genetics will likely be the method of choice for the generation of conventional influenza vaccine strains because gene reassortment by more traditional methods is cumbersome. Established systems for the artificial generation of influenza A viruses require transfection of cells with the eight to 12 plasmids that provide the eight influenza viral RNAs as well as the polymerase and nucleoproteins of the virus. However, cell lines appropriate for human vaccine production (e.g., Vero cells) cannot be transfected with high efficiencies. To overcome these problems, we established a reverse genetics system in which the eight RNA polymerase I transcription cassettes for viral RNA synthesis are combined on one plasmid. Similarly, two cassettes encoding the hemagglutinin and neuraminidase segments and six cassettes encoding the remaining proteins were combined. We also combined three RNA polymerase II transcription cassettes for the expression of the polymerase subunits. By combining these cassettes, we reduced the number of plasmids required for virus generation significantly and produced influenza A virus in Vero cells with higher efficiency than with the traditional 12 plasmid system. This new system is thus suitable for influenza virus vaccine production and may be applicable to other reverse genetics systems that rely on the introduction of several plasmids into eukaryotic cells. [ABSTRACT FROM AUTHOR]
- Subjects :
- *INFLUENZA
*VIRUS diseases
*VACCINATION
*INFLUENZA viruses
*GENETICS
*CHROMOSOMES
Subjects
Details
- Language :
- English
- ISSN :
- 00278424
- Volume :
- 102
- Issue :
- 46
- Database :
- Academic Search Index
- Journal :
- Proceedings of the National Academy of Sciences of the United States of America
- Publication Type :
- Academic Journal
- Accession number :
- 19161065
- Full Text :
- https://doi.org/10.1073/pnas.0505587102