Dolly the Sheep February 24, 1997

ELIZABETH FARNSWORTH: Seven-month-old Dolly is an ordinary-looking sheep who came into the world in an extraordinary way. She is a scientific first -- a clone -- meaning shes an exact copy of another sheep. Dollys birth last summer was the end result of 23 years of work by Dr. Ian Wilmut of the Roslin Institute just outside Edinburgh, Scotland. IAN WILMUT, Embryologist: Dolly was born last July, and shes a perfectly healthy animal. And next autumn, come the beginning of the next breeding season, shell be given the chance to breed and to continue with her normal life. ELIZABETH FARNSWORTH: Heres how Dolly came to be. Dr. Wilmut and his colleagues removed cells from the udders of a female sheep, put them in test tubes, and made them inactive or dormant. Then they removed DNA or genetic material from the unfertilized eggs of other female sheep. They inserted the DNA from the udder cells into the eggs and then exposed them to an electric current, fusing them together into one cell which could develop into an embryo. The resulting embryos were then implanted into 13 surrogate mother sheep. Only one became pregnant, and she delivered Dolly, who was named, by the way, for country western singer Dolly Parton. Cloning has long been a staple of science fiction. The dinosaurs in Jurassic Park were cloned from bits of DNA kept alive in tree sap, and in Brave New World, Aldous Huxley wrote about the production of humans and animals. And American researchers created copies of human embryos in 1993. But those embryos were not viable, and most scientists thought copying an animal was impossible until Dolly was born. Some scientists envision great benefits from cloning. They say animals could be created to produce proteins some humans lack, like the clotting factor in human blood. Cloned animals could also produce human medicines in their milk. But Wilmut drew the line at cloning humans. IAN WILMUT: We think its important to prohibit the misuses. We would share the concerns that this technique should not be used in human beings but also to make sure that we are able to use it for the purposes where there is a benefit. If the company with whom we collaborate, PPL Therapeutics, can use this to bring forward more quickly health care products for treatment of diseases like hemophilia, the blood clotting factors or the protein anti-tripsin to treat emphysema and cystic fibrosis. We think that these are important benefits which shouldnt be lost. ELIZABETH FARNSWORTH: Now well get an explanation of the cloning story from two people whove been watching it closely. Randall Prather is a professor of animal science at the University of Missouri, who is currently doing research on early embryonic development in pigs. And Paul Raeburn is a senior editor at BusinessWeek who recently authored The Last Harvest, a book about cloning animals. Thank you both for being with us. Mr. Raeburn, is this an important scientific breakthrough? PAUL RAEBURN, BusinessWeek: (New York) Its a fascinating story. I think its important on at least three levels. Its a fascinating piece of basic science. This is something that was supposed to have been impossible. Researchers thought that adult cells could not be used to produce new animals. Thats No. 1. No. 2, it has, as youve already mentioned, enormous implications, practical applications could lead to all kinds of things, new drugs, growing animal organs for transplants, and No. 3, I think itsits clear to all of us that this has tremendous mythic significance. I mean, this is just a very, very big step in making it a realistic possibility that one might think about cloning a human being. ELIZABETH FARNSWORTH: Were going get into some of that much later, a little bit later in the

show, but, Mr. Prather, I want to ask you--researchers had come fairly close to this before, right? RANDALL PRATHER, University of Missouri: (Jefferson City, MO) Yes, they have. Some of the procedures that we developed while I was at University of Wisconsin-Madison in the lab with Dr. Neil First were designed to attempt to clone embryos, not adults, but to take cells from early embryos and do the cloning procedures. If I might add to the comments that were made a little bit ago, we also have potential production agriculture, pharmaceuticals. The applications also are for research. I think thats one thing thats often omitted here. If we can create cloned animals, then we can reduce the number of animals that are needed for experimentation because, as you know, when you do experiments, you have two treatments, and the reason you have more than two animals is because of genetic variation. And by being able to do these procedures and create genetically identical individuals, hopefully, we will be able to reduce the number of animals that are needed for experimentation. ELIZABETH FARNSWORTH: Prof. Prather, lets back up a minute. I want to understand how this works. I may be old-fashioned, but I always thought you needed a sperm mixed with an egg to make the cell division occur that created an embryo. Explain how you can take DNA from an udder, put it into an egg from which the nucleus has been removed, and get life. RANDALL PRATHER: Well, in essence, the DNA in the nucleus in the udder has both the male and the female contribution because the ewe has both a mother and a father. So that genetic compatibility thats necessary is already there. The other thing that you have to do when you do the nuclear transfer is to fuse the cells. Well, when you fuse the donor cell--in this case the mammary cell with the recipient egg--you also have to trick that egg. You have to make-ELIZABETH FARNSWORTH: Okay. Back up one minute. Fuse. Just go back one minute. What do you mean by fuse? Thats where the electricity comes in, right? RANDALL PRATHER: Yes. ELIZABETH FARNSWORTH: Explain that. RANDALL PRATHER: The electric pulse does two things. One thing is it causes a transient membrane breakdown in both the donor cell and the recipient cell. When the membrane re-heals, oftentimes there are small pores that remain. These pores are thermodynamically unstable. And what will happen is those pores will enlarge just fusing the two cells together. The electric pulse does another thing. It poreates the cells such that calcium thats in the culture media can leak into the egg. That calcium leaking into the egg then mimics what the sperm does at fertilization. Abnormal fertilization, the sperm fuses with the egg, theres a huge calcium release within the egg, and so by using the electric pulse, theyre able to mimic that. ELIZABETH FARNSWORTH: Then they put the egg back into the ewe. RANDALL PRATHER: Thats correct. ELIZABETH FARNSWORTH: And only one out of thirteen actually became pregnant. Is that--do you have any idea why that happened? RANDALL PRATHER: Well, when we first began working on nuclear transfer, we did probably a thousand nuclear transfers before we got our first couple of calves. And that was using early embryonic cells, cells that havent grown through gestation and havent produced an adult animal. And we did a lot of them, and basically what that tells you is, No. 1, the procedure works. Its highly inefficient. What we really need is more research to understand exactly how its working and why it doesnt work most of the time but why it does work part of the time, so procedures do become more efficient. ELIZABETH FARNSWORTH: Paul Raeburn, why didnt somebody try this before?

PAUL RAEBURN: Well, I think there was a feeling that adult cells were irreversibly changed. What happens, as you may know, in a very early embryo after the egg has divided several times, its possible to take any one of those cells, separate it, and put it in another womb, and it can grow into another animal, another human being, if thats the case. So at a very early stage each cell has the potential to grow into the whole animal; however, as development continues, the cells lose that ability, and adult cells are believed to be, or at least were believed to be pretty much locked into their role. A liver cell was a liver cell and a skin cell was a skin cell. Now weve shown that a mammary cell can be induced to regain what it was thought-what had been thought to be irreversibly lost, that ability to regenerate and help a new egg grow into a new animal. ELIZABETH FARNSWORTH: And, Mr. Raeburn, if you can do this to an animal, can you do it to a human? PAUL RAEBURN: If you can do it to a sheep, you can do it to a human being. There may be technical problems and minor difficulties. As you noted, there were 277 eggs fertilized and put into 13 lambs to get one. That might be 2,000 it takes to get one human, or it may be simpler. But in principle, if it can be done in one mammal, it can be done in another. ELIZABETH FARNSWORTH: Randall Prather, how difficult is this process after reading the Nature article? Is it something you could do in your lab? RANDALL PRATHER: Could I do it in my lab? If--we do have the expertise, and if we had the funding and the facilities, we could probably repeat the work, yes. ELIZABETH FARNSWORTH: So its not terribly difficult, the technology? RANDALL PRATHER: I wouldnt say terribly difficult, but it does take a certain amount of expertise. ELIZABETH FARNSWORTH: And Prof. Prather, in the introductory piece before our discussion, Dr. Wilmut mentioned some of the applications, and you all have mentioned others. How would cloning lead to an animal-producing useful medicine? How does that work? RANDALL PRATHER: Well, really the next step in this process is to take those mammary epithelial cells and transect them, in other words, put in a gene of interest. If youre working with the pharmaceuticals, put in the gene that will make that pharmaceutical, select those cells that really have the gene in, functioning the way you want it to, and then use those for nuclear transfer. ELIZABETH FARNSWORTH: In other words, you take the DNA from the udders, the mammary cells, and you just put the other kinds of genes in there that you want in there? RANDALL PRATHER: Yes. ELIZABETH FARNSWORTH: Is that right? RANDALL PRATHER: Thats one logical next step, yes. ELIZABETH FARNSWORTH: So then thats not really a clone because youve added some different kinds of genes? Youre fiddling with it then? RANDALL PRATHER: Thats true. ELIZABETH FARNSWORTH: That is the scientific application. And Mr.--go ahead, sorry. RANDALL PRATHER: That is one application, yes. ELIZABETH FARNSWORTH: That is one application. And Mr. Raeburn, how about--how would it work? How would pig clones be engineered to make them good sources of organs for humans? PAUL RAEBURN: Well, when a pig organ is transplanted into a human being, or as they remember the famous story, the baboon hearts, theres a ferocious immune reaction that kills off that invading organ. What this breakthrough would enable researchers to do would be to produce a pig in which the--its normal immune system signals had been replaced by the human counterparts, so then when the organ is grown, its

still a pig heart or a pig liver, but all the signals on it that the immune system recognizes are human. So it would be recognized as a human organ when it was transplanted, which would make it feasible to do those kinds of transplants. ELIZABETH FARNSWORTH: Mr. Prather, do you have anything to add to that? Youre working with pigs. RANDALL PRATHER: Well, I might add that Im funded by the National Institutes of Health to work on those basic procedures exactly as they were described. ELIZABETH FARNSWORTH: So this may be very useful for your work, this cloning? RANDALL PRATHER: Yes, but Im not going to change my whole lab direction. We have some specific objectives that were working on and working towards very successfully, and well probably incorporate this. In fact, over the last couple of weeks weve set up a collaboration with Dr. Mark Westsuzen, and Dr. Jorge Piedraita at Texas, and to be able to do this specific procedure with pigs. ELIZABETH FARNSWORTH: And how does it work? How does the cloning help with cures for genetic diseases, or is it pretty much what youve already explained? RANDALL PRATHER: Well, its pretty much what Ive already explained. The cures really come about by understanding the basic science, and understanding the basic science is easier if the genetics are equal between the two animals that youre working on. ELIZABETH FARNSWORTH: Explain that. RANDALL PRATHER: Well, if we create genetically identical animals and we do an experiment and we can test in both animals two different treatments, then we can get a good comparison of how the animals responding, and so that these procedures will help us to understand that better. ELIZABETH FARNSWORTH: And, Mr. Raeburn, overall, what do you think is the most important of the applications? PAUL RAEBURN: Well, I think the basic science will lead to all kinds of new things that may be difficult to anticipate right now. But it--you know, it does set up the perfect nature-nurture kind of experiment to look at the effects of environment on animals. If you have two animals that are genetically identical and change their environments, theres no telling what kinds of things we might learn. So I think over the short-term drugs, organ transplants -- over the medium-term; and over the long-term its hard to say but I think tremendous advances. ELIZABETH FARNSWORTH: Thank you both very much for being with us. PAUL RAEBURN: Thank you.