US5856152A
Expired / Lapsed
US
⏱️ 0.0 years remaining
Hybrid adenovirus-AAV vector and methods of use therefor
Assignee: University of Pennsylvania
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Filing: 1994-10-28
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Granted: 1999-01-05
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Expires: 2014-10-28
Abstract
Abstract The present invention provides a hybrid vector construct which comprises a portion of an adenovirus, 5' and 3' ITR sequences from an AAV, and a selected transgene. Other hybrid vectors form a polycation conjugate and incorporate an AAV rep gene in a single particle. These hybrid virus vectors are characterized by high titer transgene delivery to a host cell and the ability to stably integrate the transgene into the host cell chromosome. Also disclosed is the use of the hybrid vectors to produce large quantities of recombinant AAV.
Inventors
- 👤 James M. Wilson
- 👤 William M. Kelley
- 👤 Krishna J. Fisher
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-11-04 03:47
CPC Classifications
C; C12; C12N; C12N15/00; C12N15/09; C12N15/63; C12N15/79; C12N15/85; C12N15/86; A; A61; A61P; A61P43/00; C07; C07K; C07K14/00; C07K14/435; C07K14/46; C07K14/47; C07K14/4701; C07K14/4712; C07K14/705; C12N15/87; A61K; A61K38/00; A61K48/00; C12N2710/00; C12N2710/00011; C12N2710/10011; C12N2710/10311; C12N2710/10341; C12N2710/10344; C12N2750/00; C12N2750/00011; C12N2750/14011; C12N2750/14111; C12N2750/14141; C12N2750/14143; C12N2750/14151; C12N2750/14152; C12N2810/00; C12N2810/40; C12N2830/00; C12N2830/008
Claims Analysis
Total Claims: 10
Claim 1
Independent Claim
125 words
A method for producing a recombinant adeno-associated virus comprising: (a) providing a hybrid viral vector comprising: (i) adenovirus sequences comprising the adenovirus 5' and 3' cis-elements necessary for replication and virion encapsidation; and (ii) adeno-associated virus sequences comprising the 5' and 3' ITRs of an adeno-associated virus, said adeno-associated virus sequences flanked by the adenoviral sequences of (i); and (iii) a selected gene operatively linked to regulatory sequences which direct expression of the selected gene in a target cell, said gene and regulatory sequences flanked by the adeno-associated virus sequences of (ii); (b) culturing a cell transfected with the vector of (a) and an optional helper virus in the presence of an AAV rep gene product; and (c) isolating from said culture a recombinant AAV.
Claim 2
Independent Claim
125 words
A method for producing a recombinant adeno-associated virus comprising: (a) providing a hybrid viral vector comprising: (i) adenovirus sequences comprising the adenovirus 5' and 3' cis-elements necessary for replication and virion encapsidation; and (ii) adeno-associated virus sequences comprising the 5' and 3' ITRs of an adeno-associated virus, said adeno-associated virus sequences flanked by the adenoviral sequences of (i); and (iii) a selected gene operatively linked to regulatory sequences which direct expression of the selected gene in a target cell, said gene and regulatory sequences flanked by the adeno-associated virus sequences of (ii); (b) culturing a cell transfected with the vector of (a) and an optional helper virus in the presence of an AAV rep gene product; and (c) isolating from said culture a recombinant AAV.
Claim 3
Dependent Claim
78 words
A hybrid viral vector comprising: (a) adenovirus sequences comprising the adenovirus 5' and 3' cis-elements necessary for replication and virion encapsidation; and (b) adeno-associated virus sequences comprising the 5' and 3' ITRs of an adeno-associated virus, said adeno-associated virus sequences flanked by the adenoviral sequences of (a); and (c) a selected gene operatively linked to regulatory sequences which direct its expression in a target cell, said gene and regulatory sequences flanked by the adeno-associated virus sequences of (b).
Claim 4
Dependent Claim
78 words
A hybrid viral vector comprising: (a) adenovirus sequences comprising the adenovirus 5' and 3' cis-elements necessary for replication and virion encapsidation; and (b) adeno-associated virus sequences comprising the 5' and 3' ITRs of an adeno-associated virus, said adeno-associated virus sequences flanked by the adenoviral sequences of (a); and (c) a selected gene operatively linked to regulatory sequences which direct its expression in a target cell, said gene and regulatory sequences flanked by the adeno-associated virus sequences of (b).
Claim 5
Dependent Claim
19 words
The vector according to claim 2 wherein said adenovirus sequences further comprise a functional deletion in the E1 gene.
Claim 6
Dependent Claim
47 words
The vector according to claim 2 wherein said adenovirus sequences further comprise functional deletions in one or more adenovirus genes selected from the group consisting of: the E2a gene, the E4 gene, the late genes L1 through L5, and the intermediate genes IX and IV a .
Claim 7
Dependent Claim
14 words
The vector according to claim 2 wherein said selected gene is a reporter gene.
Claim 8
Dependent Claim
13 words
The vector according to claim 2 further comprising an adeno-associated virus rep gene.
Claim 9
Dependent Claim
19 words
The vector according to claim 3 wherein said adenovirus sequences further comprise a functional deletion in the E3 gene.
Claim 10
Dependent Claim
27 words
The vector according to claim 5 wherein said reporter gene is selected from the group consisting of the genes encoding β-galactosidase, alkaline phosphatase and green fluorescent protein.
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USPTO Annuity & Maintenance Windows
| Annuity Stage | Window Opens | Payment Deadline | Surcharge Period Until | Estimated Fee | Status |
|---|---|---|---|---|---|
| 3.5 Years (Year 4) | 2002-01-04 | 2002-07-06 | 2003-01-05 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 2006-01-04 | 2006-07-06 | 2007-01-05 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 2010-01-04 | 2010-07-06 | 2011-01-05 | $7700.00 | Window Closed / Lapsed |