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"Fitzpatrick, Z"
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Exosome-associated AAV vector as a robust and convenient neuroscience tool
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.
Journal Article
Erratum: Exosome-associated AAV vector as a robust and convenient neuroscience tool
2016
Correction to: Gene Therapy (2016) 23, 380–392; doi:10.1038/gt.2016.11 The initial Figure 2a was erroneously generated from a file from a mouse injected with conventional AAV9-GFP and not exo-AAV9-GFP, as described in the manuscript. We have therefore reformatted this specific panel, and corrected Figure 2a and the figure legend accordingly, now showing an image of the GFP signal detected by 2-photon microscopy after intravenous injection of exo-AAV9-GFP in a mouse.
Journal Article
Mrs. John Corry Is Honored At Tallulah by Her Children
1943
As your resident director of Tallulah Falls school, I want to share with you clubwomen exciting news from the home front and to tell you one of the high moments which has come to me bringing the encouragement and inspiration I feel when I \"lift up...
Newspaper Article
LETTERS FROM THE PEOPLE
1904
Editor Constitution: Permit me to mention one or two other matters concerning Mr. Stephens, whom I knew well and intimately. I am the sole surviving member of a law class of five that boarded in the famous home of Mr.
Newspaper Article
Leading Democrats Urge Support of Nominees
1920
Editor Constitution: There is a tendency with some to bolt the ticket in regard to certain nominees of the democratic party.
Newspaper Article
Old-Time Slave Owner Asks for Fair Play
1920
Editor Constitution: Let me say a few things as regards the relations between the two races, and at the same time to commend The Constitution's attitude in behalf of fair play.
Newspaper Article
Another Smith Leader Draws the Line
1919
Editor Constitution--I had about decided to again withdraw within my shell, but after reading the grand and patriotic cards of three of my old friends, ex-Senator Perry, Judge W. A. Covington, H. H.
Newspaper Article
John A. Manget's Father Led These Brave Boys
1919
Editor Constitution: At seventy years of age I have retired after teaching schools in Blakely, Albany, Quitman and other Georgia towns for years with a driving, pushing and rushing speed and...
Newspaper Article
SAYS WILSON SIDE-STEPPED ON THE TARIFF ISSUE
1912
Editor Constitution: I am here this week attending the Chautauqua exercises--and this institution, a literary one, is the most important thing with all Albanians. I taught school hero for four years. My \"boys,\" old pupils,
Newspaper Article