Production of high-titer helper-free retroviruses by transient transfection.
- 15 September 1993
- journal article
- research article
- Published by Proceedings of the National Academy of Sciences in Proceedings of the National Academy of Sciences of the United States of America
- Vol. 90 (18), 8392-8396
- https://doi.org/10.1073/pnas.90.18.8392
Abstract
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 > 10(6) infectious 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.Keywords
This publication has 18 references indexed in Scilit:
- The Basic Science of Gene TherapyScience, 1993
- Induction of Chronic Myelogenous Leukemia in Mice by the P210
bcr/abl
Gene of the Philadelphia ChromosomeScience, 1990
- Enrichment of murine haemopoietic stem cells: diverging roadsImmunology Today, 1989
- Safe and efficient generation of recombinant retroviruses with amphotropic and ecotropic host ranges.Proceedings of the National Academy of Sciences of the United States of America, 1988
- Fluorescence-activated cell analysis and sorting of viable mammalian cells based on beta-D-galactosidase activity after transduction of Escherichia coli lacZ.Proceedings of the National Academy of Sciences of the United States of America, 1988
- Factors involved in production of helper virus-free retrovirus vectorsSomatic Cell and Molecular Genetics, 1986
- Construction of a fusion gene that confers resistance against hygromycin B to mammalian cells in cultureExperimental Cell Research, 1985
- Lethal effect of the abelson murine leukemia virus transforming gene productCell, 1981
- Characteristics of a Human Cell Line Transformed by DNA from Human Adenovirus Type 5Journal of General Virology, 1977
- Direct transformation of 3T3 cells by Abelson murine leukaemia virusNature, 1975