AAV6 Vector Production and Purification for Muscle Gene Therapy
Vectors derived from adeno-associated viruses (AAV) have been generated using numerous naturally occurring and synthetic serotypes of the virus. Such vectors have proven to be extremely useful for a variety of gene transfer studies, both in vitro and in vivo, and are increasingly being used in gene therapy protocols for a variety of human disorders. Methods to produce AAV vectors typically rely on co-transfection of several different plasmid vectors that carry the transgene of interest (the gene to be delivered , in a “transfer plasmid”) and helper genes needed for AAV vector replication and packaging (helper plasmids). While the methods used to generate AAV are conceptually simple, minor variations in a variety of steps can result in significant differences in the overall yield of vector. Here we describe protocols for generating vectors derived from AAV6, which are particularly useful for gene transfer to muscle tissues.
Key wordsAdeno-associated viral vector AAV AAV6 AAV9 Dystrophin Microdystrophin Gene therapy Systemic delivery Duchenne muscular dystrophy
This work was supported by NIH grants U54AR065139 and R01HL122332, and by Grant # 409968 from the Muscular Dystrophy Association (USA).
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