Adenovirus vectors as transcomplementing templates for the production of replication defective retroviral vectors.
We have generated an adenovirus containing a retroviral vector sequence encoding the neomycin phosphotransferase (neo) gene (AV-LXSN). AV-LXSN transduction of retroviral packaging cell lines led to production of LXSN retroviral vector with alternative viral envelopes; exposure of target cells to retroviral containing supernatants confirmed envelope specific tropism. Retroviral titers (G418 cfu/ml) were comparable to those produced by standard techniques. Retrovirus could be detected in supernatants within 24 hours of AV-LXSN transduction and persisted as long as 120 hours. Southern blot analysis of DNA purified from populations of G418 cells showed the presence of a single neo containing restriction fragment of the appropriate size that could only be generated by reverse transcription of LXSN to produce LXSN provirus. This adeno-retroviral chimeric vector system could simplify the generation and testing of different retroviral vectors, particularly where assessment of vectors with alternative envelopes carrying novel targeting ligands is required.
Ramsey
W J
WJ
Clinical Gene Therapy Branch, National Human Genome Research Institute, National Institutes of Health, Bethesda, Maryland 20892, USA. jramsey@nhgri.nih.gov
Caplen
N J
NJ
Li
Q
Q
Higginbotham
J N
JN
Shah
M
M
Blaese
R M
RM
D000256:Adenoviridae / Q000235:genetics*; D003001:Cloning, Molecular / Q000379:methods; D004340:Drug Contamination; D015316:Genetic Therapy / Q000379:methods; D005822:Genetic Vectors / Q000235:genetics*; D019868:Kanamycin Kinase / Q000235:genetics; D012367:RNA, Viral / Q000096:biosynthesis; D012190:Retroviridae / Q000235:genetics* / Q000254:growth & development / Q000472:pathogenicity; D014161:Transduction, Genetic; D019065:Virus Assembly / Q000235:genetics*
D012367:RNA, Viral; D019868:Kanamycin Kinase