期刊
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA
卷 106, 期 37, 页码 15891-15896出版社
NATL ACAD SCIENCES
DOI: 10.1073/pnas.0908897106
关键词
chimeric RNA segment; live attenuated virus; packaging signals; reverse genetics
资金
- National Institutes of Health [UO1AI070469]
- Live Attenuated Vaccines for Epidemic and Pandemic Flu [HHSN2662000700010C]
- Center for Research on Influenza Pathogenesis [U54 AI057158-04]
- Northeast Biodefense Center
Influenza viruses contain segmented, negative-strand RNA genomes. Genome segmentation facilitates reassortment between different influenza virus strains infecting the same cell. This phenomenon results in the rapid exchange of RNA segments. In this study, we have developed a method to prevent the free reassortment of influenza A virus RNAs by rewiring their packaging signals. Specific packaging signals for individual influenza virus RNA segments are located in the 5' and 3' noncoding regions as well as in the terminal regions of the ORF of an RNA segment. By putting the nonstructural protein (NS)-specific packaging sequences onto the ORF of the hemagglutinin ( HA) gene and mutating the packaging regions in the ORF of the HA, we created a chimeric HA segment with the packaging identity of an NS gene. By the same strategy, we made an NS gene with the packaging identity of an HA segment. This rewired virus had the packaging signals for all eight influenza virus RNAs, but it lost the ability to independently reassort its HA or NS gene. A similar approach can be applied to the other influenza A virus segments to diminish their ability to form reassortant viruses.
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