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V D Erdman

Publications and source records attributed to V D Erdman.

2 recordsLinked to original sources

Integration-specific retrovirus expression in embryonal carcinoma cells.

The undifferentiated embryonal carcinoma (EC) cell line PCC4 aza 1 was infected with a selectable amphotropic retrovirus. Although EC cells are generally refractory to retroviral gene expression, we found that approximately 1 of 5,000-10,000 recombinant proviruses was transcribed in these cells. Cells containing an active recombinant provirus were cloned and expanded. Nucleic acid analysis revealed approximately one intact provirus per cell. Viral RNA levels were different for each cell clone examined [between 0.05% and 0.5% of the poly(A)+ RNA], and transcription was initiated and terminated in the long terminal repeats as in permissive differentiated cells. The cells retained an undifferentiated phenotype and remained positive for stage-specific embryonic antigen 1 and negative for H-2 surface antigens. The cells retained the block to the expression of other proviruses integrated at other chromosomal locations. The data suggest that a cis-acting genetic element, at or near the chromosomal site of integration, or mutations in the viral control elements themselves may be responsible for provirus expression in these cells.

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Amphotropic retrovirus vector system for human cell gene transfer.

Retroviral vectors have been constructed for gene transfer in mammalian and avian cells, however most retroviral vector systems are complicated by the spread of a replication-competent helper virus. This problem has been circumvented by segregating the viral genome into cis- and trans-acting components. By establishing helper cell lines that produce the trans-acting viral gene products, one can propagate the cis-acting component in them and harvest defective viral particles that contain only the cis-acting component. The cis-acting component can provide a useful vehicle for the highly efficient transfer of genes into target cells. The defective vector systems described to date, however, are restricted in host range to murine, avian, rat, and dog cells. We describe a helper-free vector system based entirely on an amphotropic murine virus with a wide mammalian host range, including the ability to carry out efficient gene transfer into human cells. We also describe a double mutation constructed in the trans-acting genome which reduces the frequency of replication-competent recombinant viruses to undetectable levels.

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