Amplifiable adeno-associated virus(AAV) packaging cassettes for the production of recombinant AAV vectors
AssigneeTargeted Genetics Corporation
InventorsCarmel M. Lynch, Haim Burstein, Anthony M. Stepan, Dara H. Lockert
Life of this patent
Estimating the term…
Abstract
High-efficiency AAV packaging constructs and methods for their use are provided. in the present invention. These high-efficiency packaging constructs comprise an activating element (such as the P1 sequence located within the AAV S1 integration site of human chromosome 19) amplifiably linked to one or more AAV packaging genes. The constructs may be either integrated into a mammalian cell genome or maintained episomally. Use of the high-efficiency AAV packaging vectors of the invention provides for controlled amplifiable production of rAAV vector constructs.
Claims24
A recombinant polynucleotide comprising a sequence encoding an adeno-associated virus (AAV) packaging cassette comprising at least one AAV rep and/or cap gene amplifiably linked to a P1 element.
A method for producing high-titer stocks of an rAAV vector containing a heterologous gene of interest, comprising co-expressing the rAAV vector containing the heterologous gene of interest in a mammalian cell along with an AAV packaging cassette, said AAV packaging cassette comprising at least one AAV rep and/or cap gene amplifiably linked to a P1 element;
- and incubating the cell under conditions that allow production of high-titer stocks of the rAAV vector.
A method for generating a cell line capable of producing high-titer stocks of an rAAV vector containing a heterologous gene of interest, comprising transfecting mammalian cells with an rAAV vector containing the heterologous gene of interest and with an AAV packaging cassette, said AAV packaging cassette comprising at least one AAV rep and/or cap gene amplifiably linked to a P1 element, whereby the cell line capable of producing high-titer stocks of the rAAV vector is generated.
An AAV packaging cell line comprising mammalian cells transfected with a rAAV vector containing a heterologous gene of interest and with an AAV packaging cassette, said AAV packaging cassette comprising at least one AAV rep and/or cap gene amplifiably linked to a P1 element.
An AAV packaging cell comprising an AAV packaging cassette comprising at least one AAV rep and/or cap gene amplifiably linked to a P1 element.
Dependencies are inferred from the claim text.