Gene delivery to zebra finch (songbird) brain using AAV9-GFP vector purified with AAVpro Purification Kit Maxi (All Serotypes)

Introduction

Adeno-associated viruses (AAVs) are frequently used vectors for gene delivery to a wide variety of cell types, making for an attractive tool in gene therapies. Our AAV purification kits power in vivo experiments in laboratories worldwide, due in no small part to their ability to maintain high purity and titer.

To facilitate critical in vivo applications, Dr. Wada at Hokkaido University used one of our AAVpro "All Serotypes" purification kits for a study of the zebra finch (songbird) brain.

Results

Fluorescence of AAV9-delivered GFP was observed at injection sites

The AAVpro "all serotypes" kit was employed for purification and concentration of AAV9-GFP. Fluorescence was observed in both the song nuclei Area X and RA two weeks after the injection of 1 µl of purified AAV9-GFP vector.

Conclusions

Kit-purified AAV9-GFP vector was demonstrated to be functional following direct injection into zebra finch (songbird) brain.

Methods

Preparation of AAV vector

AAV-GFP vector was produced in HEK293 cells by introducing plasmids using the calcium phosphate transfection method. Cells cultured in ten 15-cm dishes were harvested to purify AAV-GFP particles using 1/4 of the volume specified in the protocol for the AAVpro Purification Kit Maxi (All Serotypes) (Cat. # 6666). The final volume of purified AAV-GFP vector solution was 100 µl.

Injection of AAV vector into zebra finch brain

1 µl of purified AAV9-GFP vector was injected into zebra finch (male) brain at two separate areas (song nucleus Area X* and song nucleus RA**). Brain slices were prepared two weeks after injection.

*Song nucleus Area X is equivalent to the mammalian basal ganglia.

**Song nucleus RA is equivalent to the mammalian cortical layer 5.

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