01/10/2002 08:00:00

Caltech biologists invent newer, better methodfor making transgenic animals

Using specially prepared HIV-derived viruses stripped of their disease-causing potential, California Institute of Technology biologist David Baltimore and his team have invented a new method of introducing foreign DNA into animals that could have wide-ranging applications in biotechnology and experimental biology.

The Baltimore team reports, on today's Science Express Web site, on their study of single-cell mouse embryos that have been virally infected in a manner that leaves a new gene from a jellyfish permanently deposited into their genomes. The mice, after they have been carried to term, carry at least one copy of the gene in 80 percent of the cases, and 90 percent of these show high levels of the jellyfish protein. Further, the study shows that the offspring of the mice inherit the genes and make the new protein. Thus the method makes transgenic mice that have new genetic potential.

According to Baltimore, who is president of Caltech, the use of the HIV-like viruses could prove far superior to the current method of producing transgenic animals by pronuclear injection.

"It's surprising how well it works," says Baltimore, whose Nobel Prize-winning research on the genetic mechanisms of viruses 30 years ago is central to the new technique. "This technique is much easier and more efficient than the procedure now commonly in use, and the results suggest that it can be used to generate other transgenic animal species."

The technique exploits features of HIV-like viruses known as lentiviruses, which can infect both dividing and non-dividing cells, as gene delivery vehicles. Unlike HIV, the lentivirus is rendered incapable of causing AIDS. The lentivirus carries new genes into the cell's existing genome. In this case, newly fertilized mouse eggs were engineered to carry the green fluorescent protein (GFP) derived from jellyfish.

Baltimore and his team developed two ways of introducing the lentivirus into cells: microinjection of virus under the layer that protects recently fertilized eggs, or incubation of denuded fertilized eggs in a concentrated solution of the virus. The latter method is easier, although less efficient.

The transgenic mice, once they are born, carry a protein marker in all body tissues that make them glow green under a fluorescent light. This trait is genetic because the trait is a permanent feature of the animal's genome, and thus is carried throughout life and is inheritable by offspring. The term "transgene" refers to the fact that the new gene has been transferred.

Transgenics holds promise to biotechnology and experimental biology because the techniques can be used to "engineer" new, desirable traits in plants and animals, provided the trait can be identified and localized in another organism's genome. A transgenic cow, for example, might be engineered to produce milk containing therapeutic human proteins, or a transgenic chicken might produce eggs low in cholesterol.

In experimental biology, transgenics are valuable laboratory animals for fundamental research. A cat with an altered visual system, for example, might better accommodate fundamental studies of the nature of vision.

According to Baltimore, the procedure works on rats as well as mice. This is a huge advantage to experimentalists because of the number of laboratory applications in which rats are preferable, he says.