PMID- 24222196
OWN - NLM
STAT- MEDLINE
DCOM- 20140321
LR  - 20200226
IS  - 1465-2099 (Electronic)
IS  - 0022-1317 (Linking)
VI  - 95
IP  - Pt 2
DP  - 2014 Feb
TI  - Adaptive mutation in nuclear export protein allows stable transgene expression in 
      a chimaeric influenza A virus vector.
PG  - 337-349
LID - 10.1099/vir.0.056036-0 [doi]
AB  - The development of influenza virus vectors with long insertions of foreign 
      sequences remains difficult due to the small size and instable nature of the 
      virus. Here, we used the influenza virus inherent property of self-optimization 
      to generate a vector stably expressing long transgenes from the NS1 protein ORF. 
      This was achieved by continuous selection of bright fluorescent plaques of a 
      GFP-expressing vector during multiple passages in mouse B16f1 cells. The newly 
      generated vector acquired stability in IFN-competent cell lines and in vivo in 
      murine lungs. Although improved vector fitness was associated with the appearance 
      of four coding mutations in the polymerase (PB2), haemagglutinin and 
      non-structural (NS) segments, the stability of the transgene expression was 
      dependent primarily on the single mutation Q20R in the nuclear export protein 
      (NEP). Importantly, a longer insert, such as a cassette of 1299 nt encoding two 
      Mycobacterium tuberculosis Esat6 and Ag85A proteins, could substitute for the GFP 
      transgene. Thus, the inherent property of the influenza virus to adapt can also 
      be used to adjust a vector backbone to give stable expression of long transgenes.
FAU - Kuznetsova, Irina
AU  - Kuznetsova I
AD  - Department of Surgery, Medical University of Vienna, Wahringer Gurtel 18-20, 
      A-1090, Vienna, Austria.
FAU - Shurygina, Anna-Polina
AU  - Shurygina AP
AD  - Research Institute of Influenza, Russian Academy of Medical Sciences, Moscow, 
      Russia.
FAU - Wolf, Brigitte
AU  - Wolf B
AD  - Department of Surgery, Medical University of Vienna, Wahringer Gurtel 18-20, 
      A-1090, Vienna, Austria.
FAU - Wolschek, Markus
AU  - Wolschek M
AD  - Avir Green Hills Biotechnology AG, Vienna, Austria.
AD  - Department of Surgery, Medical University of Vienna, Wahringer Gurtel 18-20, 
      A-1090, Vienna, Austria.
FAU - Enzmann, Florian
AU  - Enzmann F
AD  - Department of Surgery, Medical University of Vienna, Wahringer Gurtel 18-20, 
      A-1090, Vienna, Austria.
FAU - Sansyzbay, Abylay
AU  - Sansyzbay A
AD  - Research Institute for Biological Safety Problems, Gvardeyskyi, Kazakhstan.
FAU - Khairullin, Berik
AU  - Khairullin B
AD  - Research Institute for Biological Safety Problems, Gvardeyskyi, Kazakhstan.
FAU - Sandybayev, Nurlan
AU  - Sandybayev N
AD  - Research Institute for Biological Safety Problems, Gvardeyskyi, Kazakhstan.
FAU - Stukova, Marina
AU  - Stukova M
AD  - Research Institute of Influenza, Russian Academy of Medical Sciences, Moscow, 
      Russia.
FAU - Kiselev, Oleg
AU  - Kiselev O
AD  - Research Institute of Influenza, Russian Academy of Medical Sciences, Moscow, 
      Russia.
FAU - Egorov, Andrej
AU  - Egorov A
AD  - Research Institute of Influenza, Russian Academy of Medical Sciences, Moscow, 
      Russia.
FAU - Bergmann, Michael
AU  - Bergmann M
AD  - Comprehensive Cancer Center, Medical University of Vienna, Wahringer Gurtel 
      18-20, A-1090, Vienna, Austria.
AD  - Department of Surgery, Medical University of Vienna, Wahringer Gurtel 18-20, 
      A-1090, Vienna, Austria.
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20131112
PL  - England
TA  - J Gen Virol
JT  - The Journal of general virology
JID - 0077340
RN  - 0 (INS1 protein, influenza virus)
RN  - 0 (Recombinant Fusion Proteins)
RN  - 0 (Recombinant Proteins)
RN  - 0 (Viral Nonstructural Proteins)
RN  - 147336-22-9 (Green Fluorescent Proteins)
SB  - IM
MH  - *Active Transport, Cell Nucleus
MH  - Animals
MH  - Cell Line
MH  - *Gene Expression
MH  - Genes, Reporter
MH  - Genetic Vectors
MH  - Green Fluorescent Proteins/genetics/metabolism
MH  - Influenza A virus/*genetics/*physiology
MH  - Mice
MH  - Recombinant Fusion Proteins/genetics/metabolism
MH  - Recombinant Proteins/genetics/metabolism
MH  - Viral Load
MH  - Viral Nonstructural Proteins/*genetics/*metabolism
MH  - *Virus Replication
EDAT- 2013/11/14 06:00
MHDA- 2014/03/22 06:00
CRDT- 2013/11/14 06:00
PHST- 2013/11/14 06:00 [entrez]
PHST- 2013/11/14 06:00 [pubmed]
PHST- 2014/03/22 06:00 [medline]
AID - 10.1099/vir.0.056036-0 [doi]
PST - ppublish
SO  - J Gen Virol. 2014 Feb;95(Pt 2):337-349. doi: 10.1099/vir.0.056036-0. Epub 2013 
      Nov 12.