Metabolically activated recombinant viral vectors and methods for their preparation and use

A composition comprising a purified recombinant adeno-associated virus (rAAV) particle, wherein the rAAV particle comprises an rAAV genome comprising a heterologous nucleotide sequence comprising a coding region and one or more inverted terminal repeat (ITR) sequences flanking said heterologous sequence, wherein the total amount of unique sequence present in the heterologous sequence is about one-half of the heterologous sequence and wherein the heterologous sequence forms intrastrand base pairs along most or all of its length.

A method of introducing an rAAV genome into a cell, comprising contacting the cell essentially in the absence of an AAV helper virus with a composition comprising a purified rAAV particle under conditions that allow uptake of the rAAV vector, whereby the rAAV vector is introduced into the cell, wherein the rAAV particle comprises an rAAV genome comprising a heterologous nucleotide sequence comprising a coding region and one or more inverted terminal repeat (ITR) sequences flanking said heterologous sequence, wherein the total amount of unique sequence present in the heterologous sequence is about one-half of the heterologous sequence and wherein the heterologous sequence forms intrastrand base pairs along most or all of its length.

A method of expressing a polynucleotide coding sequence in a cell, comprising subjecting the cell to conditions which allow expression of the coding sequence, wherein the coding sequence is introduced into the cell by contacting the cell essentially in the absence of an AAV helper virus with a composition comprising a purified rAAV particle, wherein the rAAV particle comprises an rAAV genome comprising a heterologous nucleotide sequence comprising a coding region and one or more inverted terminal repeat (ITR) sequences flanking said heterologous sequence, wherein the total amount of unique sequence present in the heterologous sequence is about one-half of the heterologous sequence and wherein the heterologous sequence forms intrastrand base pairs along most or all of its length.