US 11,939,597 B2
Restrictive inverted terminal repeats for viral vectors
Curtis Hewitt, Austin, TX (US); and Richard Jude Samulski, Chapel Hill, NC (US)
Assigned to The University of North Carolina at Chapel Hill, Chapel Hill, NC (US)
Filed by The University of North Carolina at Chapel Hill, Chapel Hill, NC (US)
Filed on Dec. 1, 2022, as Appl. No. 18/060,799.
Application 18/060,799 is a division of application No. 16/953,109, filed on Nov. 19, 2020, granted, now 11,542,478.
Application 16/953,109 is a division of application No. 16/271,163, filed on Feb. 8, 2019, granted, now 10,858,632, issued on Dec. 8, 2020.
Application 16/271,163 is a division of application No. 14/922,935, filed on Oct. 26, 2015, granted, now 10,233,428, issued on Mar. 19, 2019.
Application 14/922,935 is a division of application No. 13/521,448, granted, now 9,169,494, issued on Oct. 27, 2015, previously published as PCT/US2011/020939, filed on Jan. 12, 2011.
Claims priority of provisional application 61/294,181, filed on Jan. 12, 2010.
Prior Publication US 2023/0167418 A1, Jun. 1, 2023
Int. Cl. C12N 7/00 (2006.01); C07K 14/005 (2006.01); C12N 15/86 (2006.01); C12N 15/864 (2006.01)
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.