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A system for efficient generation of adenovirus protein IX-producing helper cell lines
The journal of gene medicine, 2006-02, Vol.8 (2), p.147-154
Vellinga, Jort
Uil, Taco G.
de Vrij, Jeroen
Rabelink, Martijn J. W. E.
Lindholm, Leif
Hoeben, Rob C.
2006
Volltextzugriff (PDF)
Details
Autor(en) / Beteiligte
Vellinga, Jort
Uil, Taco G.
de Vrij, Jeroen
Rabelink, Martijn J. W. E.
Lindholm, Leif
Hoeben, Rob C.
Titel
A system for efficient generation of adenovirus protein IX-producing helper cell lines
Ist Teil von
The journal of gene medicine, 2006-02, Vol.8 (2), p.147-154
Ort / Verlag
Chichester, UK: John Wiley & Sons, Ltd
Erscheinungsjahr
2006
Quelle
MEDLINE
Beschreibungen/Notizen
Background The adenovirus 14.3 kDa hexon‐associated protein IX (pIX) functions in the viral capsid as ‘cement’ and assembles the hexons in stable groups‐of‐nine (GONs). Although viruses lacking pIX do not form GONs, and are less heat‐stable than wild‐type (wt) viruses, they can be propagated with the same kinetics and yields as the wt viruses. To facilitate ‘pseudotyping’ of adenoviral vectors we have set up an efficient system for the generation of pIX‐producing helper cell lines. Methods With a lentiviral pIX‐expression cassette, monoclonal and polyclonal helper cell lines were generated, which express wt or modified pIX genes at levels equivalent to wt HAdV‐5 infected cells. The incorporation efficiency into pIX gene deleted viruses was examined by Western analysis, immuno‐affinity electron microscopy, and heat‐stability assays. Results Immuno‐affinity electron microscopy on viruses lacking the pIX gene demonstrated that more than 96% of the particles contain pIX protein in their capsids after propagation on the pIX‐expressing helper cell lines. In addition, the pIX level in the helper cells was sufficient to generate heat‐stable particles. Finally, the ratio between pIX and fiber was equivalent to that found in wt particles. The pIX‐producing cell lines are very stable, demonstrating that pIX is not toxic to cells. Conclusion These data demonstrate that lentivirus vectors can be used for the establishment of pIX‐complementing helper cell lines. Copyright © 2005 John Wiley & Sons, Ltd.
Sprache
Englisch
Identifikatoren
ISSN: 1099-498X
eISSN: 1521-2254
DOI: 10.1002/jgm.844
Titel-ID: cdi_proquest_miscellaneous_19740986
Format
–
Schlagworte
Adenoviridae - genetics
,
Adenoviridae - metabolism
,
Adenovirus
,
Blotting, Western
,
capsid
,
Capsid Proteins - biosynthesis
,
Capsid Proteins - genetics
,
Cell Line, Transformed
,
Gene therapy
,
Genes, Reporter
,
Genetic Vectors
,
helper cells
,
Humans
,
Lentivirus
,
Microscopy, Immunoelectron
,
targeting
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