Plasmid-Driven Formation of Influenza Virus-Like Particles

ABSTRACT We established a plasmid-based system for generating infectious influenza virus-like particles entirely from cloned cDNAs. Human embryonic kidney cells (293T) were transfected with plasmids encoding the influenza A virus structural proteins and with a plasmid encoding an influenza virus-like viral RNA (vRNA) which contained an antisense copy of the cDNA for green fluorescence protein (GFP) flanked by an RNA polymerase I promoter and terminator. Intracellular transcription of the latter construct by RNA polymerase I generated GFP vRNA that was packaged into influenza virus-like particles. This system, which produced more than 104 infectious particles per ml of supernatant, would be useful in studies of influenza virus replication and particle formation. It might also benefit efforts in vaccine production and in the development of improved gene therapy vectors.

[1]  Tokiko Watanabe,et al.  Generation of influenza A viruses entirely from cloned cDNAs. , 1999, Proceedings of the National Academy of Sciences of the United States of America.

[2]  P. Gómez-Puertas,et al.  Efficient formation of influenza virus-like particles: dependence on the expression levels of viral proteins. , 1999, The Journal of general virology.

[3]  D. Pérez,et al.  The matrix 1 protein of influenza A virus inhibits the transcriptase activity of a model influenza reporter genome in vivo. , 1998, Virology.

[4]  G. Neumann,et al.  Nuclear import and export of influenza virus nucleoprotein , 1997, Journal of virology.

[5]  R. Lamb,et al.  Influenza virus hemagglutinin and neuraminidase cytoplasmic tails control particle shape , 1997, The EMBO journal.

[6]  I. Mena,et al.  Rescue of a synthetic chloramphenicol acetyltransferase RNA into influenza virus-like particles obtained from recombinant plasmids , 1996, Journal of virology.

[7]  L. Mitnaul,et al.  The cytoplasmic tail of influenza A virus neuraminidase (NA) affects NA incorporation into virions, virion morphology, and virulence in mice but is not essential for virus replication , 1996, Journal of virology.

[8]  G. Neumann,et al.  Mutational analysis of influenza virus promoter elements in vivo. , 1995, The Journal of general virology.

[9]  A. García-Sastre,et al.  The cytoplasmic tail of the neuraminidase protein of influenza A virus does not play an important role in the packaging of this protein into viral envelopes. , 1995, Virus research.

[10]  J. Rose,et al.  Novel infectious particles generated by expression of the vesicular stomatitis virus glycoprotein from a self-replicating RNA , 1994, Cell.

[11]  R. Lamb,et al.  The influenza virus hemagglutinin cytoplasmic tail is not essential for virus assembly or infectivity. , 1994, The EMBO journal.

[12]  G. Neumann,et al.  RNA polymerase I-mediated expression of influenza viral RNA molecules. , 1994, Virology.

[13]  M. Roth,et al.  Basis for selective incorporation of glycoproteins into the influenza virus envelope , 1993, Journal of virology.

[14]  Yamamura Ken-ichi,et al.  Efficient selection for high-expression transfectants with a novel eukaryotic vector , 1991 .

[15]  H. Niwa,et al.  Efficient selection for high-expression transfectants with a novel eukaryotic vector. , 1991, Gene.

[16]  A. Ishihama,et al.  RNA polymerase of influenza virus: role of NP in RNA chain elongation. , 1988, Journal of Biochemistry (Tokyo).

[17]  R. Lamb,et al.  Orthomyxoviridae: The Viruses and Their Replication. , 1996 .