PMID- 21194025
OWN - NLM
STAT- MEDLINE
DCOM- 20110708
LR  - 20121115
IS  - 1940-6029 (Electronic)
IS  - 1064-3745 (Linking)
VI  - 709
DP  - 2011
TI  - Directed evolution of adeno-associated virus (AAV) as vector for muscle gene 
      therapy.
PG  - 127-39
LID - 10.1007/978-1-61737-982-6_8 [doi]
AB  - Adeno-associated virus (AAV) is emerging as a vector of choice for muscle gene 
      therapy because of its effective and stable transduction in striated muscles. AAV 
      naturally evolve into multiple serotypes with diverse capsid gene sequences that 
      are apparently the determinants of their tissue tropism and infectivity. Certain 
      AAV serotypes show robust gene transfer upon direct intramuscular injection, 
      while others are effective in crossing the endothelial barrier to reach muscle 
      when delivered intravenously. Muscular dystrophy gene therapy requires efficient 
      body-wide muscle gene transfer. However, preferential liver transduction by 
      nearly all natural AAV serotypes could be an undesirable feature for 
      muscle-directed applications, especially by means of systemic gene delivery. Here 
      we describe a method of in vitro evolution and in vivo selection of AAV capsids 
      that target striated muscles and detarget the liver. Using DNA shuffling 
      technology, we have generated a capsid gene library by in vitro scrambling and 
      shuffling the capsid genes of natural AAV1 to AAV9. To minimize the bias and 
      limitation of in vitro screening on culture cells, we performed direct in vivo 
      panning in adult mice after intravenous injection of the shuffled capsid library 
      that packaged their own coding sequences. The AAV variants enriched in the heart 
      and muscle are retrieved by capsid gene PCR and subsequently characterized for 
      their tissue tropisms. This directed evolution and in vivo selection method 
      should be useful in generating novel gene therapy vectors for muscle and heart 
      and other tissues.
FAU - Yang, Lin
AU  - Yang L
AD  - Division of Molecular Pharmaceutics, Eshelman School of Pharmacy, University of 
      North Carolina at Chapel Hill, Chapel Hill, NC, USA.
FAU - Li, Juan
AU  - Li J
FAU - Xiao, Xiao
AU  - Xiao X
LA  - eng
GR  - AR 45967/AR/NIAMS NIH HHS/United States
GR  - AR 50595/AR/NIAMS NIH HHS/United States
PT  - Journal Article
PT  - Research Support, N.I.H., Extramural
PL  - United States
TA  - Methods Mol Biol
JT  - Methods in molecular biology (Clifton, N.J.)
JID - 9214969
RN  - 0 (Capsid Proteins)
SB  - IM
MH  - Animals
MH  - Capsid Proteins/genetics
MH  - DNA Shuffling
MH  - Dependovirus/*genetics
MH  - Directed Molecular Evolution
MH  - Gene Library
MH  - Gene Transfer Techniques
MH  - *Genetic Engineering
MH  - Genetic Therapy/*methods
MH  - Liver/metabolism
MH  - Mice
MH  - *Muscle, Striated/metabolism/physiopathology
MH  - Muscular Dystrophies/genetics/*therapy
MH  - Polymerase Chain Reaction
MH  - Recombination, Genetic
EDAT- 2011/01/05 06:00
MHDA- 2011/07/09 06:00
CRDT- 2011/01/04 06:00
PHST- 2011/01/04 06:00 [entrez]
PHST- 2011/01/05 06:00 [pubmed]
PHST- 2011/07/09 06:00 [medline]
AID - 10.1007/978-1-61737-982-6_8 [doi]
PST - ppublish
SO  - Methods Mol Biol. 2011;709:127-39. doi: 10.1007/978-1-61737-982-6_8.