Chemistry: molecular biology and microbiology – Virus or bacteriophage – except for viral vector or... – Inactivation or attenuation; producing viral subunits
Patent
1986-04-25
1989-08-29
Teskin, Robin
Chemistry: molecular biology and microbiology
Virus or bacteriophage, except for viral vector or...
Inactivation or attenuation; producing viral subunits
435 68, 435 91, 4351723, 4352402, 435320, 435948, 536 27, 935 32, 935 34, 935 57, 935 70, 935 71, C12N 704, C12N 1500, C12N 500, C12R 191
Patent
active
048617199
ABSTRACT:
DNA constructs consisting essentially of the promoter, gag, pol, and env sequences of a helper virus useful for making retrovirus packaging cell lines that do not yield helper virus and do not transfer the packaging function. Such DNA molecules are constructed by deleting from the genome of a replication-competent retrovirus all cis-acting elements except for the tRNA binding site. Specifically, deletion is made of the packaging signal, the site for initiation of second strand DNA synthesis, the site required for translation of reverse transcriptase during first strand DNA synthesis, and the provirus integration signal. DNA construct pPAM3 (ATCC No. 40234) is a representative embodiment. A cell line containing such an altered viral genome does not transmit this virus or transfer the packaging signal, but will transmit high titers of other viral RNAs containing the proper cis-acting elements, including retroviral vectors designed to carry foreign genes. Cell line PA317 (ATCC No. CLR 9078) is a representative embodiment.
REFERENCES:
patent: 4650764 (1987-03-01), Temin et al.
Mann, R., R. C. Mulligan, and D. Baltimore, 1983, Construction of a Retrovirus Packaging Mutant and Its Use to Produce Helper-Free Defective Retrovirus, Cell, 33:153-159.
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Cone, R. D., and R. C. Mulligan, 1984, High-Efficiency Gene Transfer into Mammalian Cells: Generation of Helper-Free Recombinant Retrovirus with Broad Mammalian Host Range, Proc. Natl. Acad. Sci. USA, 81:6349-6353.
Miller, A. D., M. F. Law, and I. M. Verma, 1985, Generation of Helper-Free Amphotropic Retroviruses that Transduce a Dominant-Acting Methotrexate-Resistant DHFR Gene, Mol. Cell. Biol., 5:431-437.
Sorge, J., D. Wright, V. D. Erdman, and A. E. Cutting, 1984, Amphotropic Retrovirus Vector System for Human Cell Gene Transfer, Mol. Cell. Biol., 4:1730-1737.
Panganiban, A. T., and H. M. Temin, 1983, The Terminal Nucleotides of Retrovirus DNA are Required for Integration but not Virus Production, Nature, 306:155-160.
Mann, R., and D. Baltimore, 1985, Varying the Position of a Retrovirus Packaging Sequence Results in the Encapsidation of Both Unspliced and Spliced RNAs, J. Virol, 54:401-407.
Fred Hutchinson Cancer Research Center
Peet Richard C.
Teskin Robin
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