US5124263A
Expired / Lapsed
US
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Recombination resistant retroviral helper cell and products produced thereby
Assignee: Wisconsin Alumni Research Foundation
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Filing: 1989-01-12
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Granted: 1992-06-23
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Expires: 2009-01-12
Abstract
Abstract An improved helper cell for growing up stocks of replication incompetent retrovirus vectors is disclosed. The helper cell resists recombination events due to the fact that natural promoters and poly(A) sequences in the helper sequences have been replaced with foreign promoters and poly(A) sequences bearing little or no homology to the vectors. Plasmids containing these modified sequences can still create a helper cell with resulting expression of the needed helper proteins, yet there is much less risk of recombination events in the helper cell. Also disclosed are vectors produced by such helper cells, target cells infected by such vectors, and helper cells which convert vectors so that they can be used with hosts from different species.
Inventors
- 👤 Howard M. Temin
- 👤 Joseph P. Dougherty
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-11-03 04:03
CPC Classifications
C; C12; C12N; C12N15/00; C12N15/09; C12N15/63; C12N15/79; C12N15/85; C12N15/86; C07; C07K; C07K14/00; C07K14/005; C12N7/00; C12N2740/00; C12N2740/00011; C12N2740/10011; C12N2740/13011; C12N2740/13022; C12N2740/13041; C12N2740/13043; C12N2740/13051; C12N2740/13052; C12N2740/13061; C12N2740/13062; Y; Y10; Y10S; Y10S435/00; Y10S435/8215; Y10S435/948
Claims Analysis
Total Claims: 7
Claim 1
Independent Claim
317 words
In a helper cell of the type having: a host cell; a first retrovirus helper gene sequence in the cell which has a helper portion coding for a first retrovirus virion protein for a specified retrovirus and which has a first transcriptional promoter so as to be capable of expressing the first virion protein, and a defective encapsidation portion which assists in rendering the cell by itself unable to form the infectious value which the first retrovirus helper gene sequence codes for; a second retrovirus helper gene sequence in the cell which has a helper portion coding for a second retrovirus protein for the specified retrovirus and which has a second transcriptional promoter so as to be capable of expressing the second virion protein, the second helper gene sequence also having a defective encapsidation portion which together with the defective encapsidation portion of the first helper gene sequence renders the cell by itself unable to form the infectious virus which the second retrovirus helper gene sequence codes for; a retrovirus helped gene sequence in the cell having a defective helped portion which would have rendered the cell unable to form the infectious virus which the retrovirus helped gene sequence codes for if suitable virion protein had not been supplied from expression of the first and second retrovirus helper portions or from another source; and said retrovirus helped gene sequence being a recombinant retrovirus sequence with a first part that is retrovirus and a second part that is a foreign gene protein coding part; whereby the virion proteins expressed by the first and second helper portions can permit the retrovirus helped gene sequence to form infectious recombinant virus coding for the foreign gene; the improvement comprising: no transcriptional promoter on the first or second helper sequence is present on the helped sequence and no transcriptional promoter on the first helper sequence is present on the second helper sequence.
Claim 2
Dependent Claim
317 words
In a helper cell of the type having: a host cell; a first retrovirus helper gene sequence in the cell which has a helper portion coding for a first retrovirus virion protein for a specified retrovirus and which has a first transcriptional promoter so as to be capable of expressing the first virion protein, and a defective encapsidation portion which assists in rendering the cell by itself unable to form the infectious value which the first retrovirus helper gene sequence codes for; a second retrovirus helper gene sequence in the cell which has a helper portion coding for a second retrovirus protein for the specified retrovirus and which has a second transcriptional promoter so as to be capable of expressing the second virion protein, the second helper gene sequence also having a defective encapsidation portion which together with the defective encapsidation portion of the first helper gene sequence renders the cell by itself unable to form the infectious virus which the second retrovirus helper gene sequence codes for; a retrovirus helped gene sequence in the cell having a defective helped portion which would have rendered the cell unable to form the infectious virus which the retrovirus helped gene sequence codes for if suitable virion protein had not been supplied from expression of the first and second retrovirus helper portions or from another source; and said retrovirus helped gene sequence being a recombinant retrovirus sequence with a first part that is retrovirus and a second part that is a foreign gene protein coding part; whereby the virion proteins expressed by the first and second helper portions can permit the retrovirus helped gene sequence to form infectious recombinant virus coding for the foreign gene; the improvement comprising: no transcriptional promoter on the first or second helper sequence is present on the helped sequence and no transcriptional promoter on the first helper sequence is present on the second helper sequence.
Claim 3
Dependent Claim
34 words
The helper cell of claim 1, wherein no transcriptional promoter on the first or second helper sequence has more than 35% sequence homology to the sequence of any transcriptional promoter on the helped sequence.
Claim 4
Dependent Claim
34 words
The helper cell of claim 2, wherein no 3' processing sequence on the first or second helper gene sequence has the same sequence as any 3' processing sequence present on the helped gene sequence.
Claim 5
Dependent Claim
12 words
A recombinant retrovirus vector produced by the helper cell of claim 1.
Claim 6
Dependent Claim
10 words
A host cell transformed by the vector of claim 4.
Claim 7
Dependent Claim
10 words
A progeny cell of the host cell of claim 5.
References Cited (Backward Citations)
US4650764A
Cited
US4861719A
Cited
USPTO Annuity & Maintenance Windows
| Annuity Stage | Window Opens | Payment Deadline | Surcharge Period Until | Estimated Fee | Status |
|---|---|---|---|---|---|
| 3.5 Years (Year 4) | 1995-06-23 | 1995-12-23 | 1996-06-23 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 1999-06-23 | 1999-12-23 | 2000-06-23 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 2003-06-23 | 2003-12-23 | 2004-06-23 | $7700.00 | Window Closed / Lapsed |