Production and Titering of Recombinant Adeno-associated Viral Vectors

• DOI: 10.3791/3348
• Publication Date: 2011-12-02T05:12:16Z
• Authors: Christina McClure; Katy L. H. Cole; Peer Wulff; Matthias Klugmann; Andrew J. Murray
• Keywords: Transduction (biophysics); Adeno-associated virus

In recent years recombinant adeno-associated viral vectors (AAV) have become increasingly valuable for in vivo studies animals, and are also currently being tested human clinical trials. Wild-type AAV is a non-pathogenic member of the parvoviridae family inherently replication-deficient. The broad transduction profile, low immune response as well strong persistent transgene expression achieved with these has made them popular versatile tool vitro gene delivery. rAAVs can be easily cheaply produced laboratory and, based on their favourable safety generally given classification. Here, we describe method production titering chimeric containing capsid proteins both AAV1 AAV2. use so-called combines benefits parental serotypes such high titres stocks (AAV1) purification by affinity chromatography (AAV2). These best studied all serotypes, individually infectivity pattern. described here should infectious properties AAV2 thus expected to infect large range tissues, including neurons, skeletal muscle, pancreas, kidney among others. uses heparin column purification, believed give higher titer cleaner preparation than other methods, centrifugation through caesium chloride gradient. Additionally, how quickly titered accurate reading number particles produced.