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1. WO1996039495 - AAV DIRECTED TARGETED INTEGRATION

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[ EN ]

WHAT IS CLAIMED IS:

1. A recombinant DNA vector comprising (i) AAV nucleic acid sequences that direct the integration of the recombinant DNA into regions of a host genome other than chromosome 19; and (ii) a nucleic acid sequence encoding a protein of interest.

2. A recombinant DNA vector comprising (i) non-coding AAV nucleic acid sequences that direct the integration of the recombinant DNA into regions of host genome other than chromosome 19; and (ii) a nucleic acid sequence encoding a protein of interest.

3. The recombinant DNA vector of Claim 2 wherein the AAV-noncoding nucleic acid sequences comprise (i) an AAV left ITR sequence; and (ii) an AAV right ITR sequence.

4. A method of replicating and encapsidating a recombinant DNA molecule into an AAV virion,
comprising:
a) culturing a eukaryotic cell containing
(i) a helper virus, (ii) a recombinant
nucleic acid encoding AAV REP and CAP
proteins, and (iii) the recombinant DNA
vector of claim 1, 2 or 3 whereby the
recombinant DNA vector is replicated
and assembled into AAV virions; and
(b) collecting virions produced.

5. The method according to claim 4 in which the helper virus is an adenovirus.

6. The method according to claim 4 in which the helper virus is a herpes simplex virus.

7. The method according to claim 4 in which the recombinant nucleic acid encoding the AAV REP and CAP proteins is a plasmid vector comprising:
(a) promotors which control the expression
of the AAV REP and CAP RNA;
(b) translation initiation signals for the
REP and CAP mRNA;
(c) DNA sequences encoding the REP and CAP
proteins; and
(d) transcriptional termination signals.

8. A host cell which produces stocks of
recombinant adeno-associated viruses and produced by a method comprising:
(a) culturing said host cell containing (i)
a helper virus, (ii) a recombinant
nucleic acid encoding AAV CAP and REP
proteins, and (iii) the recombinant DNA
vector of claim 1, 2 or 3 whereby the
recombinant DNA vector is replicated
and assembled into AAV virions; and
(b) collecting virions produced.

9. A mammalian genomic fragment comprising genomic nucleotide sequences, other than those located on chromosome 19, into which AAV nucleic acid
sequences are integrated.

10. The mammalian genomic fragment of claim 9 which maps to the ql2-ql3 region of chromosome 5.

11. The mammalian genomic fragment of claim 9 which maps to the pl6-p21 region of chromosome 2.

12. The mammalian genomic fragment of claim 9 which maps to the q2-q3 region of a D-group
chromosome.

13. The mammalian genomic fragment of claim 9 which maps to the pl2-pl3 region of chromosome 12.

14. The mammalian genomic fragment of claim 9 which maps to the q2 region of a D-group chromosome.

15. The mammalian genomic fragment of claim 9 which maps to the ql3 region of chromosome 22.

16. A transgenic animal, whose germ cells and somatic cells contain heterologous genomic nucleotide sequences, other than those located on human
chromosome 19, into which AAV nucleic acid sequences specifically integrate.