Directed evolution of adeno-associated virus yields enhanced gene delivery vectors.
Adeno-associated viral vectors are highly safe and efficient gene delivery vehicles. However, numerous challenges in vector design remain, including neutralizing antibody responses, tissue transport and infection of resistant cell types. Changes must be made to the viral capsid to overcome these problems; however, very often insufficient information is available for rational design of improvements. We therefore applied a directed evolution approach involving the generation of large mutant capsid libraries and selection of adeno-associated virus (AAV) 2 variants with enhanced properties. High-throughput selection processes were designed to isolate mutants within the library with altered affinities for heparin or the ability to evade antibody neutralization and deliver genes more efficiently than wild-type capsid in the presence of anti-AAV serum. This approach, which can be extended to additional gene delivery challenges and serotypes, directs viral evolution to generate 'designer' gene delivery vectors with specified, enhanced properties.
Maheshri
Narendra
N
The Department of Chemical Engineering and The Helen Wills Neuroscience Institute, The University of California, Berkeley, California 94720-1462, USA.
Koerber
James T
JT
Kaspar
Brian K
BK
Schaffer
David V
DV
D000256:Adenoviridae / Q000235:genetics*; D019020:Directed Molecular Evolution / Q000379:methods*; D016503:Drug Delivery Systems / Q000379:methods*; D018390:Gene Targeting / Q000379:methods*; D024861:Genetic Enhancement / Q000379:methods*; D015316:Genetic Therapy / Q000379:methods*; D005822:Genetic Vectors / Q000235:genetics*; D014162:Transfection / Q000379:methods*