PMID- 19888330
OWN - NLM
STAT- MEDLINE
DCOM- 20100426
LR  - 20211020
IS  - 1932-6203 (Electronic)
IS  - 1932-6203 (Linking)
VI  - 4
IP  - 11
DP  - 2009 Nov 2
TI  - AAV recombineering with single strand oligonucleotides.
PG  - e7705
LID - 10.1371/journal.pone.0007705 [doi]
LID - e7705
AB  - Adeno-associated virus (AAV) transduction initiates a signaling cascade that 
      culminates in a transient DNA damage response. During this time, host DNA repair 
      proteins convert the linear single-strand AAV genomes to double-strand circular 
      monomers and concatemers in processes stimulated by the AAV inverted terminal 
      repeats (ITRs). As the orientation of AAV genome concatemerization appears 
      unbiased, the likelihood of concatemerization in a desired orientation is low 
      (less than 1 in 6). Using a novel recombineering method, Oligo-Assisted AAV 
      Genome Recombination (OAGR), this work demonstrates the ability to direct 
      concatemerization specifically to a desired orientation in human cells. This was 
      achieved by a single-strand DNA oligonucleotide (oligo) displaying homology to 
      distinct AAV genomes capable of forming an intermolecular bridge for 
      recombination. This DNA repair process results in concatemers with genomic 
      junctions corresponding to the sequence of oligo homology. Furthermore, OAGR was 
      restricted to single-strand, not duplexed, AAV genomes suggestive of 
      replication-dependent recombination. Consistent with this process, OAGR 
      demonstrated oligo polarity biases in all tested configurations except when a 
      portion of the oligo targeted the ITR. This approach, in addition to being useful 
      for the elucidation of intermolecular homologous recombination, may find eventual 
      relevance for AAV mediated large gene therapy.
FAU - Hirsch, Matthew L
AU  - Hirsch ML
AD  - UNC Gene Therapy Center, University of North Carolina at Chapel Hill, Chapel 
      Hill, North Carolina, United States of America.
FAU - Storici, Francesca
AU  - Storici F
FAU - Li, Chengwen
AU  - Li C
FAU - Choi, Vivian W
AU  - Choi VW
FAU - Samulski, R Jude
AU  - Samulski RJ
LA  - eng
GR  - AI072176/AI/NIAID NIH HHS/United States
GR  - GM0529299/GM/NIGMS NIH HHS/United States
GR  - P01 HL066973/HL/NHLBI NIH HHS/United States
GR  - HL051818/HL/NHLBI NIH HHS/United States
GR  - HL066973/HL/NHLBI NIH HHS/United States
GR  - P01 HL051818/HL/NHLBI NIH HHS/United States
GR  - R01 AI072176/AI/NIAID NIH HHS/United States
PT  - Journal Article
PT  - Research Support, N.I.H., Extramural
PT  - Research Support, Non-U.S. Gov't
DEP - 20091102
PL  - United States
TA  - PLoS One
JT  - PloS one
JID - 101285081
RN  - 0 (DNA, Viral)
RN  - 0 (Oligonucleotides)
RN  - 147336-22-9 (Green Fluorescent Proteins)
SB  - IM
MH  - Cell Line
MH  - DNA Damage
MH  - DNA Replication
MH  - DNA, Viral
MH  - Dependovirus/*genetics
MH  - Genetic Techniques
MH  - Genetic Therapy/methods
MH  - Genetic Vectors
MH  - Genome, Viral
MH  - Green Fluorescent Proteins/metabolism
MH  - Humans
MH  - Models, Genetic
MH  - Oligonucleotides/*genetics
MH  - *Recombination, Genetic
MH  - Signal Transduction
PMC - PMC2765622
COIS- Competing Interests: The authors have declared that no competing interests exist.
EDAT- 2009/11/06 06:00
MHDA- 2010/04/27 06:00
PMCR- 2009/11/02
CRDT- 2009/11/06 06:00
PHST- 2009/06/29 00:00 [received]
PHST- 2009/09/28 00:00 [accepted]
PHST- 2009/11/06 06:00 [entrez]
PHST- 2009/11/06 06:00 [pubmed]
PHST- 2010/04/27 06:00 [medline]
PHST- 2009/11/02 00:00 [pmc-release]
AID - 09-PONE-RA-11318R1 [pii]
AID - 10.1371/journal.pone.0007705 [doi]
PST - epublish
SO  - PLoS One. 2009 Nov 2;4(11):e7705. doi: 10.1371/journal.pone.0007705.