US 5,843,742 A · Adeno-associated derived vector systems for gene delivery and integration into target cells

Abstract

A novel nucleotide sequence integration and targeting system is described. The system employs adeno-associated virus (AAV) derived vectors which include a selected nucleotide sequence bounded by AAV inverted terminal repeats (ITRs). An AAV rep coding region is also provided, enabling the targeted integration of the selected nucleotide sequence into the genome of a suitable target cell. The nucleotide sequence integration system of the present invention can deliver and integrate large segments of DNA into the genome of target cells. Further, the subject integration system provides the advantage of site-specific integration of the selected nucleotide sequences in a target cell genome, thereby avoiding insertional mutagenesis events experienced with prior systems.

Claims40

Dependent claims

A method of integrating a selected nucleotide sequence into the genome of a mammalian cell, comprising:

(a) providing:
(i) a mammalian cell;
(ii) a first virion-free nucleic acid construct comprising a nucleotide sequence flanked by a 5' and a 3' adeno-associated virus inverted terminal repeat;
and
(iii) a second virion-free nucleic acid construct having a rep coding region operably linked to control elements which direct the transcription and translation of the rep coding region in said mammalian cell;
(b) introducing, in any order, said first and second virion-free nucleic acid constructs into said mammalian cell;
and
(c) expressing the rep coding region of said second nucleic acid construct, thereby producing an amount of rep expression product under conditions such that an increased frequency of integration of said nucleotide sequence of said first nucleic acid construct into the genome of said mammalian cell is achieved.

5 more elements

Dependencies are inferred from the claim text.

Adeno-associated derived vector systems for gene delivery and integration into target cells

Life of this patent

Adeno-associated derived vector systems for gene delivery and integration into target cells

Life of this patent

Abstract

Claims40