Efficient propagation of single gene deleted recombinant Sendai virus vectors.
Recombinant Sendai virus vectors (SeVV) have become an attractive tool for basic virological as well as for gene transfer studies. However, to (i) reduce the cellular injury induced by basic recombinant SeV vectors (encoding all six SeV genes as being present in SeV wild-type (wt) genomes) and to (ii) improve SeV vector safety, deletions of viral genes are necessary for the construction of superior SeVV generations. As a strong expression system recombinant replication-incompetent adenoviruses, coding for SeV proteins hemagglutinin-neuraminidase (HN), fusion (F), or matrix (M), were generated and successfully employed for the propagation of single gene deleted (DeltaHN, DeltaF, DeltaM) recombinant SeVV. Further investigations of the propagation procedures required for single gene deleted recombinant SeVV demonstrated (i) modifications of the cell culture medium composition as well as (ii) incubation with vitamin E as crucial steps for the enhancement of SeVV-DeltaHN, -DeltaF, or -DeltaM viral particle yield. Such optimized propagation procedures even led to a successful propagation of HN-deleted viral particles (SeVV-DeltaHN), which has not been reported before.
Bernloehr
Christian
C
Internal Medicine I, University Clinic Tübingen, D-72076, Tübingen, Germany.
Bossow
Sascha
S
Ungerechts
Guy
G
Armeanu
Sorin
S
Neubert
Wolfgang J
WJ
Lauer
Ulrich M
UM
Bitzer
Michael
M
D000256:Adenoviridae / Q000235:genetics / Q000502:physiology; D000818:Animals; D002522:Chlorocebus aethiops; D003470:Culture Media; D017353:Gene Deletion; D005814:Genes, Viral; D005816:Genetic Complementation Test; D005822:Genetic Vectors; D006378:Helper Viruses / Q000235:genetics / Q000502:physiology; D029082:Sendai virus / Q000235:genetics* / Q000502:physiology*; D014709:Vero Cells; D014779:Virus Replication; D014810:Vitamin E / Q000494:pharmacology
D003470:Culture Media; D014810:Vitamin E