Highly productive packaging lines
AssigneeUniversite Pierre et Marie Curie
InventorsDavid Klatzmann, Jean-Loup Salzmann
Life of this patent
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Abstract
The invention concerns a packaging eukaryotic cell for the production of defective infections viruses carrying a transgene, characterized in that it is deficient in one cell function essential to its growth, in particular in the presence of a selection culture medium, the function being capable of being restored by the expression of an exogenous sequence introduced in the cell: either with a vector carrying transcomplementing functions of packaging cells; or with a vector carrying a transgene, and enabling the selection in a selective medium of cells carrying the said exogenous sequence.
Claims11
A packaging cell line for the production of defective infectious retroviruses, wherein said packaging cell line is a human cell comprising, in its genome, at least two retroviral vectors:
- (a) wherein the first retroviral vector comprises, under the control of a LTR (Long Terminal Repeat) type promoter, the sequence of retroviral gag/pol genes, a mutated or normal IRES (Internal Ribosome Entry Sites) sequence, the sequence of a first selection gene, and a polyadenylation sequence;
- (b) wherein the second retroviral vector comprises, under the control of a promoter, the sequence of a gene coding for an envelope protein, a mutated or normal IRES sequence, the sequence of a second selection gene and a polyadenylation sequence;
- and
- (c) said first and second vectors being free of a retroviral packaging sequence.
A packaging cell line for the production of defective infectious retroviruses, wherein said packaging cell line is a human cell comprising, in its genome, at least a first and a second viral constructs,
- said first viral construct comprising, operably linked, a promoter, the sequence of retroviral gag/pol genes, a mutated or normal IRES (Internal Ribosome Entry Sites) sequence, the sequence of a first selection gene and a polyadenylation sequence,
- said second viral construct comprising, operably linked, a promoter, the sequence of a gene coding for an envelope protein, a mutated or normal IRES sequence, the sequence of a second selection gene and a polyadenylation sequence,
- said first and second constructs being free of a retroviral packaging sequence.
A packaging cell line for the production of defective infectious viruses, wherein said packaging cell line is a human cell comprising, in its genome, at least a first a second viral constructs,
- said first viral construct comprising, operably linked, a retroviral LTR (Long Terminal Repeat) type promoter sequence, the sequence of retroviral gag/pol genes, a mutated or normal IRES (Internal Ribosome Entry Sites) sequence, the sequence of a first selection gene selected from the group consisting of a blasticidine S resistance gene (BSR gene) and a zeomycine resistance gene (ZeoR gene), and a polyadenylation sequence,
- said second viral construct comprising, operably linked, a cytomegalovirus (CMV) promoter, the sequence of a gene coding for an envelope protein, a mutated or normal IRES sequence, the sequence of a phleomycine resistance gene and a polyadenylation sequence, said first and second constructs being free of a retroviral packaging sequence.
Dependencies are inferred from the claim text.