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Original Article – Enabling Technologies

Exosome-associated AAV vector as a robust and convenient neuroscience tool

A Corrigendum to this article was published on 03 November 2016


Adeno-associated virus (AAV) vectors are showing promise in gene therapy trials and have proven to be extremely efficient biological tools in basic neuroscience research. One major limitation to their widespread use in the neuroscience laboratory is the cost, labor, skill and time-intense purification process of AAV. We have recently shown that AAV can associate with exosomes (exo-AAV) when the vector is isolated from conditioned media of producer cells, and the exo-AAV is more resistant to neutralizing anti-AAV antibodies compared with standard AAV. Here, we demonstrate that simple pelleting of exo-AAV from media via ultracentrifugation results in high-titer vector preparations capable of efficient transduction of central nervous system (CNS) cells after systemic injection in mice. We observed that exo-AAV is more efficient at gene delivery to the brain at low vector doses relative to conventional AAV, even when derived from a serotype that does not normally efficiently cross the blood–brain barrier. Similar cell types were transduced by exo-AAV and conventionally purified vector. Importantly, no cellular toxicity was noted in exo-AAV-transduced cells. We demonstrated the utility and robustness of exo-AAV-mediated gene delivery by detecting direct GFP fluorescence after systemic injection, allowing three-dimensional reconstruction of transduced Purkinje cells in the cerebellum using ex vivo serial two-photon tomography. The ease of isolation combined with the high efficiency of transgene expression in the CNS, may enable the widespread use of exo-AAV as a neuroscience research tool. Furthermore, the ability of exo-AAV to evade neutralizing antibodies while still transducing CNS after peripheral delivery is clinically relevant.

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We thank Dr Miguel Sena-Esteves for providing the AAV plasmids as well as Dr David P. Corey for the expertise and resources of his laboratory on the cochlear transduction analysis. This work was supported by NIH/NINDS R21 NS081374-01 (CM), an American Brain Tumor Association Discovery grant (CM) and the Alzheimer’s Drug Discovery Foundation (EH) and JPB Foundation (BTH). In addition, this work was also partially supported by NIH/NINDS R01 NS086570-01 (SHR). We would like to acknowledge the Nucleic Acid Quantitation Core at MGH Neuroscience Center for qPCR analysis.

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Correspondence to C A Maguire.

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CM has a patent application related to the Exo-AAV (vexosome) technology and has received royalty payments from agreements between Partners Healthcare and Exosome Diagnostics, Inc. CM has received an honorarium from Biogen Idec for giving a seminar on the technology. The other authors declare no conflict of interest.

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Hudry, E., Martin, C., Gandhi, S. et al. Exosome-associated AAV vector as a robust and convenient neuroscience tool. Gene Ther 23, 380–392 (2016).

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