PMID- 9261406
OWN - NLM
STAT- MEDLINE
DCOM- 19970917
LR  - 20200724
IS  - 0022-538X (Print)
IS  - 1098-5514 (Electronic)
IS  - 0022-538X (Linking)
VI  - 71
IP  - 9
DP  - 1997 Sep
TI  - Identification and elimination of replication-competent adeno-associated virus 
      (AAV) that can arise by nonhomologous recombination during AAV vector production.
PG  - 6816-22
AB  - Adeno-associated virus (AAV) vector preparations are often contaminated with 
      variable amounts of replication-competent AAV (rcAAV), which may influence the 
      behavior of these vectors both in cultured cells and in animals. A packaging 
      plasmid/vector plasmid system containing no significant homology and lacking the 
      wild-type AAV p5 promoter was constructed to eliminate the production of 
      wild-type AAV by recombination. Still, rcAAV was detected in vector produced by 
      cotransfection of these plasmids at large scale. Sequence analysis revealed that 
      nonhomologous recombination was responsible for the generation of these novel 
      rcAAVs. A new AAV packaging plasmid carrying separate rep and cap expression 
      cassettes in opposite transcriptional orientations was constructed. AAV vector 
      preparations produced by using this packaging construct did not contain rcAAV.
FAU - Allen, J M
AU  - Allen JM
AD  - Fred Hutchinson Cancer Research Center, Seattle, Washington 98109, USA.
FAU - Debelak, D J
AU  - Debelak DJ
FAU - Reynolds, T C
AU  - Reynolds TC
FAU - Miller, A D
AU  - Miller AD
LA  - eng
GR  - CA09229/CA/NCI NIH HHS/United States
GR  - DK47754/DK/NIDDK NIH HHS/United States
GR  - HL53570/HL/NHLBI NIH HHS/United States
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PT  - Research Support, U.S. Gov't, P.H.S.
PL  - United States
TA  - J Virol
JT  - Journal of virology
JID - 0113724
RN  - 0 (DNA-Binding Proteins)
RN  - 0 (Viral Proteins)
RN  - 137750-19-7 (rep proteins, Adeno-associated virus 2)
RN  - 9038-94-2 (Metallothionein)
SB  - IM
MH  - Animals
MH  - Capsid/genetics
MH  - Cell Line, Transformed
MH  - Cytomegalovirus/genetics
MH  - DNA-Binding Proteins/genetics
MH  - Dependovirus/genetics/*isolation & purification/physiology
MH  - *Genetic Vectors
MH  - Humans
MH  - Metallothionein/genetics
MH  - Mice
MH  - Recombination, Genetic
MH  - Transformation, Genetic
MH  - Tumor Cells, Cultured
MH  - Viral Proteins/genetics
MH  - *Virus Replication
PMC - PMC191962
EDAT- 1997/09/01 00:00
MHDA- 1997/09/01 00:01
PMCR- 1997/09/01
CRDT- 1997/09/01 00:00
PHST- 1997/09/01 00:00 [pubmed]
PHST- 1997/09/01 00:01 [medline]
PHST- 1997/09/01 00:00 [entrez]
PHST- 1997/09/01 00:00 [pmc-release]
AID - 10.1128/JVI.71.9.6816-6822.1997 [doi]
PST - ppublish
SO  - J Virol. 1997 Sep;71(9):6816-22. doi: 10.1128/JVI.71.9.6816-6822.1997.