US4980289A
Expired / Lapsed
US
⏱️ 0.0 years remaining
Promoter deficient retroviral vector
Assignee: Wisconsin Alumni Research Foundation
•
Filing: 1987-04-27
•
Granted: 1990-12-25
•
Expires: 2007-04-27
Abstract
Abstract A recombinant retrovirus vector is disclosed. It is of the type having a normally replication incompetent retrovirus gene sequence with a foreign eukaryotic gene to be expressed. The retrovirus gene sequence is designed so as to be promoter deficient in the right side LTR. The vector can produce progeny virus from helper cells, which progeny can infect a eukaryotic host cell, form a provirus, and express the eukaryotic gene in the host cell. However, the provirus will then be defective in the retrovirus promoter, such that retrovirus RNA is not expressed from the provirus.
Inventors
- 👤 Howard M. Temin
- 👤 Joseph P. Dougherty
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-10-30 09:50
CPC Classifications
C; C12; C12N; C12N15/00; C12N15/09; C12N15/63; C12N15/79; C12N15/85; C12N15/86; C12N2740/00; C12N2740/00011; C12N2740/10011; C12N2740/13011; C12N2740/13041; C12N2740/13043
Claims Analysis
Total Claims: 8
Claim 1
Independent Claim
212 words
In a recombinant retrovirus vector having a 5' LTR, a 3' LTR, a promoter that can be recognized by a selected eukaryotic host cell, and a non-retroviral gene under the control of said recognized promoter, an improvement comprising: the 5' LTR is positioned 5' of the non-retroviral gene and has a transcriptional promoter sequence and a viral integration point; the 3' LTR is positioned 3' of the non-retroviral gene and has a viral integration point, a defective polyadenylation assistance sequence portion in the U3 region and a defective transcriptional promoter portion; an exogenous polyadenylation addition signal sequence recognized by the selected host cell is positioned on the vector 3' to the 3' LTR viral integration point; and the recognized promoter is positioned adjacent to the non-retroviral gene on the vector so as to permit expression of the non-retroviral gene in the eukaryotic host cell; whereby when constructed as described above the vector can still produce progeny virus in a helper cell with the progeny virus being capable of infecting the selected eukaryotic host cell and forming a provirus in the host cell, with the non-retroviral gene then being expressible in the host cell, but the provirus in the host cell will be replication incompetent even in the presence of a helper virus.
Claim 2
Dependent Claim
212 words
In a recombinant retrovirus vector having a 5' LTR, a 3' LTR, a promoter that can be recognized by a selected eukaryotic host cell, and a non-retroviral gene under the control of said recognized promoter, an improvement comprising: the 5' LTR is positioned 5' of the non-retroviral gene and has a transcriptional promoter sequence and a viral integration point; the 3' LTR is positioned 3' of the non-retroviral gene and has a viral integration point, a defective polyadenylation assistance sequence portion in the U3 region and a defective transcriptional promoter portion; an exogenous polyadenylation addition signal sequence recognized by the selected host cell is positioned on the vector 3' to the 3' LTR viral integration point; and the recognized promoter is positioned adjacent to the non-retroviral gene on the vector so as to permit expression of the non-retroviral gene in the eukaryotic host cell; whereby when constructed as described above the vector can still produce progeny virus in a helper cell with the progeny virus being capable of infecting the selected eukaryotic host cell and forming a provirus in the host cell, with the non-retroviral gene then being expressible in the host cell, but the provirus in the host cell will be replication incompetent even in the presence of a helper virus.
Claim 3
Dependent Claim
297 words
In a recombinant retrovirus vector having a 5' LTR, a 3' LTR, a promoter that can be recognized by a selected eukaryotic host cell, and a non-retroviral gene under the control of said recognized promoter, an improvement comprising: the 5' LTR is positioned 5' of the non-retroviral gene and has a transcriptional promoter sequence and a viral integration point; the 3' LTR is positioned in 3' of the non-retroviral gene and has a viral integration point, a defective polyadenylation assistance sequence portion, and a defective transcriptional promoter portion; a first exogenous polyadenylation addition signal sequence recognized by the selected host cell is positioned on the vector 3' to the 3' LTR viral integration point; a second exogenous polyadenylation addition signal sequence recognized by the selected host cell is positioned on the vector; the recognized promoter is positioned adjacent to the non-retroviral gene on the vector so as to permit expression of the non-retroviral gene in the eukaryotic host cell; and the transcriptional orientation of the recognized promoter, non-retroviral gene, and second exogenous polyadenylation addition signal sequence is inverted relative to the transcriptional orientation of the 5' LTR transcriptional promoter and first exogenous polyadenylation addition signal sequence, and the second exogenous polyadenylation addition signal sequence is adjacent to the non-retroviral gene at the end of the non-retroviral gene that is opposite the end to which the recognized promoter is adjacent; whereby when constructed as described above the vector can still produce progeny virus in a helper cell with the progeny virus being capable of infecting the selected eukaryotic host cell and forming a provirus in the host cell, with the non-retroviral gene then being expressible in the host cell, but the provirus in the host cell will be replication incompetent even in the presence of a helper virus.
Claim 4
Dependent Claim
297 words
In a recombinant retrovirus vector having a 5' LTR, a 3' LTR, a promoter that can be recognized by a selected eukaryotic host cell, and a non-retroviral gene under the control of said recognized promoter, an improvement comprising: the 5' LTR is positioned 5' of the non-retroviral gene and has a transcriptional promoter sequence and a viral integration point; the 3' LTR is positioned in 3' of the non-retroviral gene and has a viral integration point, a defective polyadenylation assistance sequence portion, and a defective transcriptional promoter portion; a first exogenous polyadenylation addition signal sequence recognized by the selected host cell is positioned on the vector 3' to the 3' LTR viral integration point; a second exogenous polyadenylation addition signal sequence recognized by the selected host cell is positioned on the vector; the recognized promoter is positioned adjacent to the non-retroviral gene on the vector so as to permit expression of the non-retroviral gene in the eukaryotic host cell; and the transcriptional orientation of the recognized promoter, non-retroviral gene, and second exogenous polyadenylation addition signal sequence is inverted relative to the transcriptional orientation of the 5' LTR transcriptional promoter and first exogenous polyadenylation addition signal sequence, and the second exogenous polyadenylation addition signal sequence is adjacent to the non-retroviral gene at the end of the non-retroviral gene that is opposite the end to which the recognized promoter is adjacent; whereby when constructed as described above the vector can still produce progeny virus in a helper cell with the progeny virus being capable of infecting the selected eukaryotic host cell and forming a provirus in the host cell, with the non-retroviral gene then being expressible in the host cell, but the provirus in the host cell will be replication incompetent even in the presence of a helper virus.
Claim 5
Dependent Claim
12 words
A retrovirus that has been produced by the vector of claim 1.
Claim 6
Dependent Claim
12 words
A retrovirus that has been produced by the vector of claim 2.
Claim 7
Dependent Claim
9 words
A host cell containing the provirus of claim 1.
Claim 8
Dependent Claim
9 words
A host cell containing the provirus of claim 2.
References Cited (Backward Citations)
US4650764A
Cited
USPTO Annuity & Maintenance Windows
| Annuity Stage | Window Opens | Payment Deadline | Surcharge Period Until | Estimated Fee | Status |
|---|---|---|---|---|---|
| 3.5 Years (Year 4) | 1993-12-24 | 1994-06-25 | 1994-12-25 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 1997-12-24 | 1998-06-25 | 1998-12-25 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 2001-12-24 | 2002-06-25 | 2002-12-25 | $7700.00 | Window Closed / Lapsed |