Claims for Patent: 7,060,497

Summary for Patent: 7,060,497

Title:	Adeno-associated viral vector-based methods and compositions for introducing an expression cassette into a cell
Abstract:	Methods and compositions are provided for introducing an expression cassette into a cell. In the subject methods, a population of at least two distinct adeno-associated viral particles is provided, where each distinct type of viral particle in the population comprises a different portion of the expression cassette to be introduced into the cell. The target cell is contacted with population of adeno-associated viral vectors under conditions sufficient to produce a hetero-concatemer in the cell, where the hetero-concatemer includes a functional expression cassette having an intron that includes an ITR sequence. Also provided by the subject invention are vector preparations for practicing the subject methods as well as kits for use in producing the vectors employed in the subject methods. The subject methods find use in a variety of different gene transfer applications, including both in vivo and in vitro gene transfer applications, and are particularly suited for use in the transfer of long genes.
Inventor(s):	Nakai; Hiroyuki (Redwood City, CA), Kay; Mark A. (Los Altos, CA)
Assignee:	The Board of Trustees of the Leland Stanford Junior University (Palo Alto, CA)
Application Number:	09/797,858
Patent Claims:	1. A method for introducing a functional expression cassette into a cell, said method comprising: (a) providing a population of first and second adeno-associated viral particles, wherein said first adeno-associated viral particle comprises a first portion of an expression cassette and said second adeno-associated viral particle comprises a second portion of an expression cassette; and (b) contacting said cell with said first and second adeno-associated viral particles under conditions sufficient for a hetero-concatemer to be produced inside said cell, wherein said hetero-concatemer comprises a functional expression cassette made up of said first and second portions separated by an intron comprising an inverted terminal repeat (ITR) sequence, wherein said contacting occurs in vitro; to introduce a functional expression cassette into said cell. 2. The method according to claim 1, wherein said functional expression cassette comprises DNA capable of being transcribed into an mRNA molecule. 3. The method according to claim 1, wherein said functional expression cassette is longer than about 4.7 kb. 4. The method according to claim 1, wherein said population comprises only two distinct adeno-associated viral vectors. 5. A kit for use in introducing a functional expression cassette into a cell, said kit comprising: first and second plasmid vectors for producing a population of at least two distinct adeno-associated viral particles, wherein each distinct member of said population of at least two distinct adeno-associated viral particles comprises a different portion of said single an expression cassette that produce a hetero-concatemer comprising a single expression cassette; and instructions for practicing the method according to claim 1. 6. The kit according to claim 5, wherein said first plasmid vector comprises an insertion site flanked by adeno-associated virus inverted terminal repeat (ITR) sequences. 7. The kit according to claim 5, wherein said second plasmid vector comprises Rep and Cap coding sequences. 8. The kit according to claim 5, wherein said kit further comprises a source of adenovirus helper proteins. 9. The kit according to claim 5, wherein said kit further further comprises a packaging cell line. 10. An isolated cell comprising a an adeno-associated viral hetero-concatemer, wherein said hetero-concatemer comprises a functional expression cassette, wherein said functional expression cassette comprises at least two portions of the functional expression cassette separated by an intron comprising an inverted terminal repeat (ITR) sequence. 11. The isolated cell according to claim 10, wherein said expression cassette is longer than about 4.7 kb. 12. The isolated cell according to claim 10, wherein said expression cassette comprises two portions separated by an intron comprising an ITR sequence. 13. The isolated cell according to claim 12, wherein said cell is a hepatic cell. 14. An that produce a hetero-concatemer comprising a single expression cassette adeno-associated viral vector preparation for introducing an expression cassette into a cell, comprising: a population of at least two distinct adeno-associated viral particles, wherein each distinct member of said population comprises a different portion of said single expression cassette. 15. The preparation according to claim 14, wherein said expression cassette is capable of being transcribed into an mRNA. 16. The preparation according to claim 14, wherein said expression cassette is longer than about 4.7 kb in length. 17. The preparation according to claim 14, wherein said preparation comprises a first population of adeno-associated viral vectors comprising a first portion of said expression cassette and a second population of viral vectors comprising a second portion of said expression cassette. 18. The preparation according to claim 14, wherein said preparation is present in a pharmaceutical delivery vehicle.

Details for Patent 7,060,497

Applicant	Tradename	Biologic Ingredient	Dosage Form	BLA	Approval Date	Patent No.	Expiredate
Merck Sharp & Dohme Corp.	INTRON A	interferon alfa-2b	For Injection	103132	06/04/1986		2020-03-03
Merck Sharp & Dohme Corp.	INTRON A	interferon alfa-2b	For Injection	103132			2020-03-03
Merck Sharp & Dohme Corp.	INTRON A	interferon alfa-2b	Injection	103132			2020-03-03