United States Patent:  6,086,913

Assignee:  University of British Columbia (Burnaby, CA).

Filed:  Oct. 25, 1996

Abstract

The invention provides compositions and methods for introducing a nucleic acid fragment into the genome of a cell. Suitable compositions comprise an expression vector having first and second inverted repeated sequences from an adeno associated virus, a rep gene from an adeno associated virus and the nucleic acid fragment. The expression vector is complexed with lipids.

SUMMARY OF THE INVENTION

The invention provides compositions and methods for introducing a nucleic acid fragment into the genome of a cell. Such a composition comprises an expression vector comprising first and second inverted repeated sequences from an adeno associated virus, a rep gene from an adeno associated virus and the nucleic acid fragment. The expression vector is complexed with lipids. Optionally, the vector further comprises at least one gene encoded by adenovirus DNA segments E1-E4, and some vectors include each of these segments. Optionally, the vector includes a loxP site. Often, the nucleic acid fragment encodes a protein coding sequence in operable linkage to a promoter. Alternatively, the fragment can encode an antisense transcript.

The invention further provides methods of modifying the phenotype of cells, comprising contacting the cells with compositions as described above. Often, the cells to be modified have a mutant gene associated with a defective expression product. The DNA fragment encodes a functional expression product of the gene and the gene integrates into the genome of the cells and is expressed. Optionally, the composition further comprises a pharmaceutical excipient. In some methods, the cells are from a patient suffering from a disease resulting from the mutant form of the gene, and the method further comprising reintroducing the cells into the patient. In some methods, the recipient cells are incapable of replication. In some methods, the nucleic acid fragment integrates into about 5% of the recipient cells. In some methods, the cells are contacted with the composition in vivo in a patient.

Claim 1 of 13 Claims

1. A serum-stable composition for introducing a nucleic acid fragment into the genome of a cell, comprising:

an expression vector comprising first and second inverted repeated sequences from an adeno associated virus, a rep gene from an adeno associated virus and the nucleic acid fragment; and

wherein the expression vector is in a lipid formulation.

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