US5139941A
Expired / Lapsed
US
⏱️ 0.0 years remaining
AAV transduction vectors
Assignee: University of Florida Research Foundation Inc
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Filing: 1991-10-25
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Granted: 1992-08-18
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Expires: 2011-10-25
Abstract
Abstract A hybrid gene vector suitable for introducing foreign DNA into a mammalian cell comprising the foreign DNA ligated to an AAV genome; a method of constructing the hybrid gene vector; a method of transducing foreign DNA into mammalian cells comprising infecting the cells with the above hybrid gene vector and a method of rescuing foreign DNA from mammalian cells utilizing helper virus.
Inventors
- 👤 Nicholas Muzyczka
- 👤 Paul L. Hermonat
- 👤 Kenneth I. Berns
- 👤 Richard J. Samulski
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-11-03 01:59
CPC Classifications
C; C12; C12N; C12N15/00; C12N15/09; C12N15/63; C12N15/79; C12N15/85; C12N15/86; C12N2750/00; C12N2750/00011; C12N2750/14011; C12N2750/14111; C12N2750/14141; C12N2750/14143
Claims Analysis
Total Claims: 13
Claim 1
Independent Claim
151 words
A hybrid gene vector comprising foreign DNA ligated into an AAV genome in place of or in addition to the cap, lip or rep coding sequence thereof or in place of or in addition to an AAV DNA sequence excluding the first and last 145 base pairs, said vector being capable of transducing data foreign DNA into a mammalian cell in the presence of a promoter other than an AAV transcription promoter, and said AAV genome being cloned in a prokaryotic vector plasmid or bacteriophage vector capable of growth in prokaryotic cells or yeast plasmid vector capable of growth in fungal cells and isolating a recombinant AAV hybrid gene vector by transfection of the ligated vector into a mammalian cell, said AAV genome being the entire AAV viral DNA or a deletion mutant thereof, capable of replication upon infection of said mammalian cell when complemented in trans by an AAV gene.
Claim 2
Dependent Claim
151 words
A hybrid gene vector comprising foreign DNA ligated into an AAV genome in place of or in addition to the cap, lip or rep coding sequence thereof or in place of or in addition to an AAV DNA sequence excluding the first and last 145 base pairs, said vector being capable of transducing data foreign DNA into a mammalian cell in the presence of a promoter other than an AAV transcription promoter, and said AAV genome being cloned in a prokaryotic vector plasmid or bacteriophage vector capable of growth in prokaryotic cells or yeast plasmid vector capable of growth in fungal cells and isolating a recombinant AAV hybrid gene vector by transfection of the ligated vector into a mammalian cell, said AAV genome being the entire AAV viral DNA or a deletion mutant thereof, capable of replication upon infection of said mammalian cell when complemented in trans by an AAV gene.
Claim 3
Dependent Claim
12 words
The vector of claim 1 wherein said AAV genome is AAV-type 2.
Claim 4
Dependent Claim
16 words
The vector of claim 2 wherein said AAV genome is the deletion mutant dl52-91 or dl3-94.
Claim 5
Independent Claim
155 words
A method of constructing a hybrid gene vector comprising ligating foreign DNA into an AAV genome in place of or in addition to the cap, lip or rep coding sequence thereof or in place of or in addition to an AAV DNA sequence excluding the first and last 145 base pairs, said vector being capable of transducing said foreign DNA into a mammalian cell in the presence of a promoter other than an AAV transcription promoter, and said AAV genome being cloned in a prokaryotic vector plasmid or bacteriophage vector capable of growth in prokaryotic cells or yeast plasmid vector capable of growth in fungal cells and isolating a recombinant AAV hybrid gene vector by transfection of the ligated vector into a mammalian cell, said AAV genome being the entire AAV viral DNA or a deletion mutant thereof, capable of replication upon infection of said mammalian cell when complemented in trans by an AAV gene.
Claim 6
Independent Claim
155 words
A method of constructing a hybrid gene vector comprising ligating foreign DNA into an AAV genome in place of or in addition to the cap, lip or rep coding sequence thereof or in place of or in addition to an AAV DNA sequence excluding the first and last 145 base pairs, said vector being capable of transducing said foreign DNA into a mammalian cell in the presence of a promoter other than an AAV transcription promoter, and said AAV genome being cloned in a prokaryotic vector plasmid or bacteriophage vector capable of growth in prokaryotic cells or yeast plasmid vector capable of growth in fungal cells and isolating a recombinant AAV hybrid gene vector by transfection of the ligated vector into a mammalian cell, said AAV genome being the entire AAV viral DNA or a deletion mutant thereof, capable of replication upon infection of said mammalian cell when complemented in trans by an AAV gene.
Claim 7
Dependent Claim
14 words
The method of claim 4 wherein said AAV genome is an AAV-type 2 genome.
Claim 8
Dependent Claim
16 words
The method of claim 5 wherein said AAV genome is the deletion mutant dl52-91 or dl13-94.
Claim 9
Dependent Claim
17 words
The method of claim 4 wherein said AAV genome is cloned in the prokaryotic plasmid vector pBR322.
Claim 10
Dependent Claim
42 words
The method of claim 4 wherein said transfection is conducted in the presence of a helper virus and/or a helper AAV plasmid ins 96/λ-M each of which contain genes necessary for the replication of AAV genome or the production of AAV virus.
Claim 11
Independent Claim
165 words
A method of transducing foreign DNA into mammalian cells comprising infecting said cells with a hybrid gene vector comprising foreign DNA ligated into an AAV genome in place of or in addition to the cap, lip or rep coding sequence thereof or in place of or in addition to an AAV DNA sequence excluding the first and last 145 base pairs, said vector being capable of transducing said foreign DNA into a mammalian cell in the presence of a promoter other than an AAV transcription promoter, and said AAV genome being cloned in a prokaryotic vector plasmid or bacteriophage vector capable of growth in prokaryotic cells or yeast plasmid vector capable of growth in fungal cells and isolating a recombinant AAV hybrid gene vector by transfection of the ligated vector into a mammalian cell, said AAV genome being the entire AAV viral DNA or a deletion mutant thereof, capable of replication upon infection of said mammalian cell when complemented in trans by an AAV gene.
Claim 12
Independent Claim
165 words
A method of transducing foreign DNA into mammalian cells comprising infecting said cells with a hybrid gene vector comprising foreign DNA ligated into an AAV genome in place of or in addition to the cap, lip or rep coding sequence thereof or in place of or in addition to an AAV DNA sequence excluding the first and last 145 base pairs, said vector being capable of transducing said foreign DNA into a mammalian cell in the presence of a promoter other than an AAV transcription promoter, and said AAV genome being cloned in a prokaryotic vector plasmid or bacteriophage vector capable of growth in prokaryotic cells or yeast plasmid vector capable of growth in fungal cells and isolating a recombinant AAV hybrid gene vector by transfection of the ligated vector into a mammalian cell, said AAV genome being the entire AAV viral DNA or a deletion mutant thereof, capable of replication upon infection of said mammalian cell when complemented in trans by an AAV gene.
Claim 13
Dependent Claim
42 words
A method of rescuing foreign DNA from mammalian cells transduced according to a method of claim 9 comprising infecting said transduced mammalian cells with helper virus containing genes necessary for the replication of the AAV genome or the production of AAV virus.
References Cited (Backward Citations)
US4797368A
Cited
USPTO Annuity & Maintenance Windows
| Annuity Stage | Window Opens | Payment Deadline | Surcharge Period Until | Estimated Fee | Status |
|---|---|---|---|---|---|
| 3.5 Years (Year 4) | 1995-08-18 | 1996-02-17 | 1996-08-18 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 1999-08-18 | 2000-02-17 | 2000-08-18 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 2003-08-18 | 2004-02-17 | 2004-08-18 | $7700.00 | Window Closed / Lapsed |