The spread of a replication-competent MuLV retroviral vector can be efficiently blocked by deletion variants.
A retroviral vector in which the gag and pol genes have been replaced by the NLS-lacZ reporter gene was derived from a cloned AKV-like virus. A complementing cell line expressing the gag and pol retroviral genes was constructed. The retroviral vector was demonstrated to replicate in the complementing cells. Since transfection is known to generate deletion variants of the introduced plasmid, we have examined whether it can give rise to viral forms with a replicating advantage over the initial vector. After transfection in complementing cells the spread of the vector was followed by X-gal staining. The fraction of stained cells increased for the first 10 days following transfection and was then stabilized to about 20% stained cells, thus defining two cell types; one with LacZ+ phenotype and one with LacZ- phenotype. Molecular analysis showed that the latter contains a deleted form of the virus preventing cell infection by the vector presumably through a mechanism of interference involving the viral env gene. Thus, interference results in the efficient block of vector expansion.
Nouvel
P
P
Unité de Génétique des Mammifères, Institut Pasteur, Paris, France.
Panthier
J J
JJ
Condamine
H
H
D001483:Base Sequence; D002461:Cell Line, Transformed; D003673:Defective Viruses / Q000235:genetics* / Q000502:physiology; D015685:Fusion Proteins, gag-pol / Q000235:genetics; D015752:Genes, env / Q000235:genetics; D005822:Genetic Vectors; D009052:Leukemia Virus, Murine / Q000235:genetics*; D008969:Molecular Sequence Data; D016366:Open Reading Frames; D011533:Proviruses / Q000235:genetics; D012367:RNA, Viral / Q000032:analysis; D017384:Sequence Deletion / Q000502:physiology*; D014158:Transcription, Genetic; D014162:Transfection; D014779:Virus Replication / Q000235:genetics*; D001616:beta-Galactosidase / Q000235:genetics
D015685:Fusion Proteins, gag-pol; D012367:RNA, Viral; D001616:beta-Galactosidase