PMID- 12539150
OWN - NLM
STAT- MEDLINE
DCOM- 20040219
LR  - 20061115
IS  - 1099-498X (Print)
IS  - 1099-498X (Linking)
VI  - 5
IP  - 2
DP  - 2003 Feb
TI  - Analysis of site-specific transgene integration following cotransduction with 
      recombinant adeno-associated virus and a rep encodingplasmid.
PG  - 120-9
AB  - BACKGROUND: Recombinant adeno-associated virus (rAAV) has many advantages for 
      gene therapeutic applications in comparison with other vector systems. One of the 
      most promising features is the ability of wild-type (wt) AAV to integrate 
      site-specifically into human chromosome 19. However, this feature is lost in rAAV 
      vectors due to the removal of the rep-coding sequences. METHODS: HeLa cells were 
      transfected with a rep expression plasmid, infected by rAAV and grown with or 
      without selection pressure. Single cell clones were generated and genomic DNA was 
      analyzed for site-specific integration by Southern blotting analysis and 
      fluorescence in situ hybridization (FISH). RESULTS: Transfection of HeLa cells 
      with a rep expression plasmid followed by transduction with a rAAV vector 
      resulted in site-specific integration of the transgene at AAVS1 on human 
      chromosome 19 in 7 of 10 cell clones analyzed. In marked contrast, transduction 
      of cells with rAAV alone did not result in any site-specific integration of the 
      transgene. CONCLUSIONS: The high frequency with which the site-specific 
      integration took place in the presence of Rep protein is comparable with the 
      results observed with wtAAV. These results offer opportunities for the 
      development of specifically integrating rAAV vectors.
CI  - Copyright 2002 John Wiley & Sons, Ltd.
FAU - Huttner, Nadja A
AU  - Huttner NA
AD  - Gene Center of the Ludwig-Maximilians-University Munich, Feodor-Lynen-Str. 25, 
      81377 Munich, Germany.
FAU - Girod, Anne
AU  - Girod A
FAU - Schnittger, Susanne
AU  - Schnittger S
FAU - Schoch, Claudia
AU  - Schoch C
FAU - Hallek, Michael
AU  - Hallek M
FAU - Buning, Hildegard
AU  - Buning H
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - England
TA  - J Gene Med
JT  - The journal of gene medicine
JID - 9815764
RN  - 0 (DNA-Binding Proteins)
RN  - 0 (Recombinant Fusion Proteins)
RN  - 0 (Viral Proteins)
RN  - 137750-19-7 (rep proteins, Adeno-associated virus 2)
SB  - IM
MH  - Adenoviridae/*genetics
MH  - Chromosomes, Human, Pair 19/genetics/metabolism
MH  - DNA-Binding Proteins/genetics/*metabolism
MH  - HeLa Cells
MH  - Humans
MH  - In Situ Hybridization, Fluorescence
MH  - Plasmids/*genetics
MH  - Recombinant Fusion Proteins/genetics/metabolism
MH  - *Transduction, Genetic
MH  - *Transgenes
MH  - Viral Proteins/genetics/*metabolism
EDAT- 2003/01/23 04:00
MHDA- 2004/02/20 05:00
CRDT- 2003/01/23 04:00
PHST- 2003/01/23 04:00 [pubmed]
PHST- 2004/02/20 05:00 [medline]
PHST- 2003/01/23 04:00 [entrez]
AID - 10.1002/jgm.324 [doi]
PST - ppublish
SO  - J Gene Med. 2003 Feb;5(2):120-9. doi: 10.1002/jgm.324.