Safe and efficient generation of recombinant retroviruses with amphotropic and ecotropic host ranges.

来自 NCBI

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100

作者：

Danos，O.，Mulligan，R.，C.

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摘要：

We have constructed a set of packaging cell lines useful for the generation of helper-free recombinant retroviruses with amphotropic and ecotropic host ranges. To eliminate the problems of transfer of packaging functions and helper virus formation encountered with the previously available packaging systems, two mutant Moloney murine leukemia virus-derived proviral genomes carrying complementary mutations in the gag-pol or env regions were sequentially introduced into NIH 3T3 cells by cotransformation. Both genomes contained a deletion of the ψ sequence necessary for the efficient encapsidation of retroviral genomes into virus particles and additional alterations at the 3′ end of the provirus. We show that the resulting packaging cell lines ψ CRIP and ψ CRE can be used to isolate clones that stably produce high titers (106 colony-forming units/ml) of recombinant retroviruses with amphotropic and ecotropic host ranges, respectively. More importantly, we demonstrate that viral producers derived from the packaging cell lines do not transfer the packaging functions, or yield helper virus, even under conditions where existing packaging cell lines can be shown to yield transfer of packaging functions and/or helper virus. These properties of the ψ CRIP and ψ CRE packaging lines make them particularly valuable reagents for in vivo gene transfer studies aimed at cell lineage analysis and the development of human gene replacement therapies.

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关键词：

Genetics Retroviral Vectors Packaging Cell Lines Gene Transfer

DOI：

10.1073/pnas.85.17.6460

被引量：

2230

年份：

1988