From sniffles and pinkeye to acute respiratory disease and hemorrhagic cystitis, adenoviruses have a lot to answer for. But recombinant adenoviruses, stripped of their infectivity, have a lot to offer as gene-therapy vectors of choice. They can transfer genes to a variety of cell types, with high levels of protein production.
Yet the generation of such multi-use gene delivery vehicles in eukaryotic host cells is fraught with difficulty and tedium. Now molecular biologist Bert Vogelstein of the Howard Hughes Medical Institute at Johns Hopkins University, in Baltimore, has come up with a better way.
His article in the March 2, 1998, issue of the Proceedings of the National Academy of Sciences bears the title: "A simplified system for generating recombinant adenoviruses."
"Basically," Vogelstein told BioWorld Today, "it employs a method in which the homologous recombination is done in bacteria rather than in mammalian cells. Its advantages include reproducibility, as well as simplicity. Bacterial recombination," he pointed out, "is much more efficient and much easier to verify that the correct recombinants have been produced."
The second improvement, Vogelstein added, "is that a reporter gene expressing green fluorescent protein (GFP) is included in the vector backbone. This greatly simplifies following the whole process of viral production, which is historically one of the most difficult, and often frustrating, parts of the process."
He explained: "At all times you have a quite visual striking demonstration of how viral production is proceeding, and how infection is progressing — both during the production process and subsequently, during use of the virus that's produced."
While assuming that his new method is patentable, Vogelstein made the point: "Our goal is to have this used as widely as possible, if other investigators find it as convenient and useful as we have done. Several people have already written us and requested viruses."
Simplifying this process too, Vogelstein supplied his e-mail address: firstname.lastname@example.org. — David N. Leff