PUBLICATION
Generation of high-titer pseudotyped retroviral vectors with very broad host range
- Authors
- Yee, J.K., Friedmann, T., and Burns, J.C.
- ID
- ZDB-PUB-970102-13
- Date
- 1994
- Source
- Methods in cell biology 43: 99-112 (Other)
- Registered Authors
- Burns, Jane C.
- Keywords
- none
- MeSH Terms
-
- 3T3 Cells
- Animals
- Base Sequence
- Cell Line
- Gene Transfer Techniques
- Genetic Vectors*/physiology
- Humans
- Insecta
- Mammals
- Membrane Glycoproteins*
- Mice
- Molecular Sequence Data
- Moloney murine leukemia virus/genetics*
- Moloney murine leukemia virus/physiology
- Polymerase Chain Reaction
- Rats
- Vesicular stomatitis Indiana virus/physiology*
- Viral Envelope Proteins/physiology*
- Xenopus
- Zebrafish
- PubMed
- 7823872 Full text @ Meth. Cell. Biol.
Citation
Yee, J.K., Friedmann, T., and Burns, J.C. (1994) Generation of high-titer pseudotyped retroviral vectors with very broad host range. Methods in cell biology. 43:99-112.
Abstract
Encapsidation of the VSV G protein into the virions of MoMLV- derived retroviral vectors in the absence of other VSV-encoded proteins is shown to be an efficient process, although the exact mechanism for this process is currently unclear. Unlike the conventional retroviral vectors bearing the amphotropic envelope protein, the pseudotyped virus has the ability to withstand the shearing forces encountered during ultracentrifugation. This property of the pseudotyped virus enables the generation of high-titer retroviral vector stocks and has potential application for in vivo gene therapy studies. We have found as many as four copies of a pseudotyped vector to integrate into the genome of a single cell when a high multiplicity of infection was used to infect the cells. Multiple integration events were not observed with amphotropic retroviral vectors, probably because of their low virus titers. In addition, when retroviral vectors are pseudotyped with the VSV G protein, they acquire the host range of VSV and are able to infect nonmammalian cells derived from fish, Xenopus, mosquito, and Lepidoptera. Since techniques for efficient gene transfer in some of these nonmammalian systems are not currently available, retrovirus-mediated gene transfer described here should be useful for transgenic and other genetic studies in lower vertebrate species. The inability to establish a stable cell line expressing the VSV G protein, however, limits large-scale production of the pseudotyped retroviral vectors. Generation of stable packaging cell lines for the pseudotyped retroviral vectors is a major challenge for the future.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping