CPC C12N 7/00 (2013.01) [C07K 14/005 (2013.01); C12N 15/86 (2013.01); C12N 15/8645 (2013.01); C12N 2750/14122 (2013.01); C12N 2750/14143 (2013.01); C12N 2750/14162 (2013.01); C12N 2750/14322 (2013.01); C12N 2750/14352 (2013.01); C12N 2820/60 (2013.01)] | 28 Claims |
1. A method of producing a recombinant parvovirus particle, comprising providing to a cell permissive for parvovirus replication:
A) a recombinant parvovirus template, comprising:
i) a heterologous nucleic acid; and
ii) at least one parvovirus inverted terminal repeat (ITR), wherein said ITR comprises:
a) a first structural element that functionally interacts with a large Rep protein from a first adeno-associated virus (AAV) but does not functionally interact with a large Rep protein from a second AAV; and
b) a second structural element that functionally interacts with the large Rep protein from the second AAV but does not functionally interact with the large Rep protein from the first AAV;
wherein the ITR functionally interacts with a synthetic AAV large Rep protein; and
B) a polynucleotide encoding a synthetic large Rep protein comprising a first portion that functionally interacts with a first structural element of a parvovirus ITR and a second portion that functionally interacts with a second structural element of a parvovirus ITR, wherein said first structural element functionally interacts with a large Rep protein from a first AAV but does not functionally interact with a large Rep protein from a second AAV and said second structural element functionally interacts with a large Rep protein from a second AAV but does not functionally interact with a large Rep protein from the first AAV;
under conditions sufficient for the replication and packaging of the recombinant parvovirus template;
whereby recombinant parvovirus particles are produced in the cell.
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