Production of High-Titer Helper-Free Retroviruses by Transient Transfection

The generation of high-titer, helper-free retroviruses by transient transfection has been achieved by using the highly transfectable 293T cell line into which are stably introduced constructs that express retroviral packaging functions. The resulting ecotropic virus packaging cell line BOSC 23 produ...

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Bibliographic Details
Published in:Proceedings of the National Academy of Sciences - PNAS 1993-09, Vol.90 (18), p.8392-8396
Main Authors: Pear, Warren S., Nolan, Garry P., Scott, Martin L., Baltimore, David
Format: Article
Language:English
Subjects:
3T3 Cells
Animals
beta-Galactosidase - biosynthesis
beta-Galactosidase - metabolism
Biological and medical sciences
Bone Marrow Transplantation - physiology
Cell Line
Cell lines
Cell Transformation, Neoplastic
Cells
Cellular biology
Chloroquine - pharmacology
Female
Fundamental and applied biological sciences. Psychology
Genes, abl
Genetic engineering
HEK293 cells
Helper Viruses - physiology
Humans
Male
Medical research
Mice
Mice, Inbred C57BL
Microbiology
NIH 3T3 cells
Packaging
Plasminogen Activators - biosynthesis
Repetitive Sequences, Nucleic Acid
Retroviral vectors
Retroviridae
Retroviridae - drug effects
Retroviridae - genetics
Retroviridae - physiology
RNA-Directed DNA Polymerase - analysis
Techniques used in virology
Transfection
Virology
Virus Replication - drug effects
Viruses
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Description
Summary:The generation of high-titer, helper-free retroviruses by transient transfection has been achieved by using the highly transfectable 293T cell line into which are stably introduced constructs that express retroviral packaging functions. The resulting ecotropic virus packaging cell line BOSC 23 produces infectious retrovirus at >106infectious units/ml of supernatant within 72 hr after CaPO4-mediated transfection. A stringent assay for replication-competent virus showed that no helper virus was present. The system can produce high titers of retroviral vectors expressing genes that are extremely difficult to propagate at high titer in stable producer lines. This method should facilitate and extend the use of helper-free retroviral gene transfer, as well as be useful for gene therapy.
ISSN:0027-8424
1091-6490
DOI:10.1073/pnas.90.18.8392