The First Adeno-Associated Virus Gene Transfer Experiment,1983

Abstract

The first recombinant adeno-associated virus (AAV) gene delivery/gene therapy experiment was the delivery of the neomycin resistance gene by AAV2 into Detroit 6 cells. Figure 1 shows the first experiment as well as the resulting cellular phenotype of G418-resistant cell colonies. This experiment was carried out in 1983, and the results were published within a table in a 1984 publication (Hermonat and Muzyczka, 1984). However, the photograph was never published.

FIG. 1.

The first adeno-associated virus (AAV) gene transfer experiment. This first experiment was carried out in 1983, and the results were published in a table in a 1984 publication (Hermonat and Muzyczka, 1984). However, this photograph was never published until now. An AAV/Neo-transduced plate is on the left (A), and a mock-transduced plate is on the right (B). Both plates were then G-418 selected.

The AAV2/Neo virus was generated using the earliest production system composed of infecting cells with wild-type adenovirus 5 and transfection of two AAV plasmids. One was the AAV/Neo vector plasmid, and the other plasmid contained a large wild-type AAV2 genome too large to be packaged. This large wild-type AAV provided all the missing AAV products in trans for the AAV/Neo vector (Hermonat and Muzyczka, 1984).

Because this was a landmark experiment, and we had no knowledge as to the potential dangers of AAV gene transfer, this experiment was carried out in the University of Florida's P3 facility. I was dressed in full personal protective equipment—full gown, gloves, and respirator—while carrying out this experiment. In this experiment, one plate was infected with AAV/Neo virus (multiplicity of infection of 1,000) and another plate was mock transduced. After transduction of the human Detroit 6 cells, 1 mg/ml of G418 was added 7 days after infection.

As can be seen, the results were dramatic, and thus we knew, even in 1983, that AAV would be a very successful gene transfer tool. From this first experiment and publication on AAV gene transfer (Hermonat and Muzyczka, 1984), there are now 2,500 articles on the topic (PubMed search). Additionally, 1983–1984 were very exciting years for the birth of gene therapy with, in addition to the first AAV gene transfer publication, the first retroviral and adenoviral gene transfer studies also being published (Joyner and Bernstein, 1983; Cone and Mulligan, 1984; Van Doren et al., 1984).

Footnotes

Acknowledgments

The author wishes to acknowledge Nick Muzyczka, Kenneth Berns, and all the University of Florida faculty and students at that time for their support.

Author Disclosure Statement

No competing financial interests exist.

References

Cone

R.D.

, and Mulligan

R.C.

(1984). High efficiency gene transfer into mammalian cells: generation of helper-free recombinant retrovirus with broad mammalian host range. Proc. Natl. Acad. Sci. USA, 81, 6349–6353.

Hermonat

P.L.

, and Muzyczka

(1984). Use of adeno-associated virus as a mammalian DNA cloning vector: transduction of neomycin resistance into mammalian tissue culture cells. Proc. Natl. Acad. Sci. USA, 81, 6466–6470.

Joyner

A.L.

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(1983) Retrovirus transduction: generation of infectious retroviruses expressing dominant and selectable genes is associated with in vivo recombination and deletion events. Mol. Cell Biol., 3, 2180–2190.

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, Hanahan

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(1984) Infection of eukaryotic cells by helper-independent recombinant adenoviruses: early region 1 is not obligatory for integration of viral DNA. J. Virol., 50, 606–614.