US 2003/0073232 A1 · Methods and vector constructs useful for production of recombinant AAV

Abstract

Methods for efficient production of recombinant AAV employ a host cell containing a first nucleic acid molecule comprising from 5′ to 3′, a parvovirus P5 promoter, a spacer, an AAV rep sequence and an AAV cap gene sequence, wherein said spacer is of sufficient size to reduce expression of the rep78 and rep68 gene products; a second nucleic acid molecule comprising a minigene comprising a transgene flanked by AAV inverse terminal repeats (ITRs) and under the control of regulatory sequences directing expression thereof in a host cell; and helper functions essential to the replication and packaging of rAAV, which functions are not provided by the first or second nucleic acid molecules. Host cells and molecule constructs are also described.

Claims17

Dependent claims

A recombinant host cell containing

(a) a first nucleic acid molecule comprising a parvovirus P5 promoter, an AAV rep gene sequence, an AAV cap gene sequence, and a spacer of sufficient size to reduce expression of the rep78 and rep68 gene products relative to other rep gene products, wherein said spacer is heterologous to said promoter and is interposed between said promoter and the start site of the rep gene, and
(b) a second nucleic acid molecule comprising a minigene comprising a transgene flanked by AAV inverted terminal repeats (ITRs) and under the control of regulatory sequences directing expression thereof in a host cell;
and
(c) helper functions essential to the replication and packaging of rAAV.

1 more element

Dependencies are inferred from the claim text.

Methods and vector constructs useful for production of recombinant AAV

Life of this patent

Methods and vector constructs useful for production of recombinant AAV

Life of this patent

Abstract

Claims17