PMID- 18728640
OWN - NLM
STAT- MEDLINE
DCOM- 20081212
LR  - 20240416
IS  - 1525-0024 (Electronic)
IS  - 1525-0016 (Print)
IS  - 1525-0016 (Linking)
VI  - 16
IP  - 10
DP  - 2008 Oct
TI  - DNA shuffling of adeno-associated virus yields functionally diverse viral 
      progeny.
PG  - 1703-9
LID - 10.1038/mt.2008.167 [doi]
AB  - Adeno-associated virus (AAV) vectors are extremely effective gene-delivery 
      vehicles for a broad range of applications. However, the therapeutic efficacy of 
      these and other vectors is currently limited by barriers to safe, efficient gene 
      delivery, including pre-existing antiviral immunity, and infection of off-target 
      cells. Recently, we have implemented directed evolution of AAV, involving the 
      generation of randomly mutagenized viral libraries based on serotype 2 and 
      high-throughput selection, to engineer enhanced viral vectors. Here, we 
      significantly extend this capability by performing high-efficiency in vitro 
      recombination to create a large (10(7)), diverse library of random chimeras of 
      numerous parent AAV serotypes (AAV1, 2, 4-6, 8, and 9). In order to analyze the 
      extent to which such highly chimeric viruses can be viable, we selected the 
      library for efficient viral packaging and infection, and successfully recovered 
      numerous novel chimeras. These new viruses exhibited a broad range of cell 
      tropism both in vitro and in vivo and enhanced resistance to human intravenous 
      immunoglobulin (IVIG), highlighting numerous functional differences between these 
      chimeras and their parent serotypes. Thus, directed evolution can potentially 
      yield unlimited numbers of new AAV variants with novel gene-delivery properties, 
      and subsequent analysis of these variants can further extend basic knowledge of 
      AAV biology.
FAU - Koerber, James T
AU  - Koerber JT
AD  - Department of Chemical Engineering, University of California at Berkeley, 
      Berkeley, California 94720-1462, USA.
FAU - Jang, Jae-Hyung
AU  - Jang JH
FAU - Schaffer, David V
AU  - Schaffer DV
LA  - eng
GR  - R01 HL081527/HL/NHLBI NIH HHS/United States
GR  - R01 HL081527-02/HL/NHLBI NIH HHS/United States
PT  - Journal Article
PT  - Research Support, N.I.H., Extramural
PT  - Research Support, Non-U.S. Gov't
PT  - Research Support, U.S. Gov't, Non-P.H.S.
DEP - 20080826
PL  - United States
TA  - Mol Ther
JT  - Molecular therapy : the journal of the American Society of Gene Therapy
JID - 100890581
RN  - 0 (DNA Primers)
RN  - 0 (DNA, Viral)
SB  - IM
MH  - Animals
MH  - Base Sequence
MH  - Cell Line
MH  - DNA Primers
MH  - *DNA Shuffling
MH  - DNA, Viral/*genetics
MH  - Dependovirus/*genetics/physiology
MH  - Directed Molecular Evolution
MH  - Female
MH  - Humans
MH  - Mice
MH  - Mice, Inbred BALB C
MH  - Neutralization Tests
MH  - Polymerase Chain Reaction
MH  - Transduction, Genetic
MH  - Virus Assembly
PMC - PMC2683895
MID - NIHMS109873
EDAT- 2008/08/30 09:00
MHDA- 2008/12/17 09:00
PMCR- 2009/05/18
CRDT- 2008/08/30 09:00
PHST- 2008/08/30 09:00 [pubmed]
PHST- 2008/12/17 09:00 [medline]
PHST- 2008/08/30 09:00 [entrez]
PHST- 2009/05/18 00:00 [pmc-release]
AID - S1525-0016(16)32074-3 [pii]
AID - 10.1038/mt.2008.167 [doi]
PST - ppublish
SO  - Mol Ther. 2008 Oct;16(10):1703-9. doi: 10.1038/mt.2008.167. Epub 2008 Aug 26.