Adeno-associated viral Particle (AAV)

For generating adeno-associated viruses you can choose from 3 shuttle vector types. All contain 5’ and 3’ AAV ITRs and a WPRE. In the basic vector you can express the transgene by using a variety of different promoters. In the moment we are using CamKII, hSynapsinI, CMV early enhancer/chicken β actin (CAG), phosphoglycerate kinase (PGK), EF-1 alpha, CMV and a liver specific promoter (LP1).

In the second set of shuttle vectors we apply LoxP sequences and an invert-oriented transgene to allow specific expression in Cre-transgenic animals. Here the same promoter settings are available as in the first vector type.

In the third set of AAV shuttle vectors we apply the hSynapsin1 promoter together with reporter proteins coupled via a Öffnet externen Link im aktuellen Fenster2A Sequenz to a MCS into which the transgene can be cloned. This configuration allows a bicistronic expression of reporter gene and transgene through one promoter.

We utilize helper plasmids providing adenoviral genes necessary to drive AAV replication (E2a, E4, VARNA). In order to achieve tissue specific expression we are applying specific AAV serotype helper plasmids which envelope the AAV capsid for the Serotypes 1,2,3,4,5,6,8,9 and rh10. The plasmids express AAV2 replicase and the capsid specific proteins.

The underlying purification method depends on the serotype of the generated AAV. If you wish, heparin-columns are used for AAV2/2 particle purification. Iodixanol gradiends are used for the others. If necessary HPLC purification methods are used for AAV2/8 (in cooperation with AG Weger, Institute of Virology, Charité).

External users needs specific MTAs for the plasmids provided by Penn Vector Core (for serotype 9 and rh10). More informations to this matter the user will get within the Opens external link in current windowOrdering Prozesses.

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