And a happy new year to all. --PG http://www.nytimes.com/2009/01/01/garden/01clones.html January 1, 2009 Living Together Beloved Pets Everlasting? By ERIC KONIGSBERG Fairfax, Calif. THE most difficult thing about the cloned puppies is not telling them apart, but explaining why they don't look exactly alike. This was the problem Lou Hawthorne faced on a recent afternoon hike with Mira and MissyToo, two dogs whose embryos were created from the preserved, recycled and repurposed nuclear DNA of the original Missy, a border collie-husky mix who died in 2002. To be sure, they have a very strong resemblance to each other and to Missy. It's just that sometimes, as soon as people hear that the dogs are clones, the questions start coming: "Why is one dog's fur curlier?" "Why aren't the dogs the same size?" "Why is one of them darker?" "Why does this one have a floppy ear?" Mr. Hawthorne, who is 48, is highly invested in the notion of likeness. With clones, after all, what good does similar do? It is Mr. Hawthorne's biotech company, BioArts, which is based here in the Bay Area but has arrangements with a laboratory in South Korea, that performed the actual cloning. He also has particular reason to be sensitive to questions that touch on the authenticity of the clones, given the history of his chief geneticist, Dr. Hwang Woo Suk of the Sooam Biotech Research Foundation in South Korea. Dr. Hwang is perhaps best known for fraudulently reporting in 2004 that a team he led had successfully cloned human embryos and stem cells. After the false claims were unearthed, he was fired by Seoul National University, where he did his research as a professor. But he is also widely acknowledged for having been involved in successfully cloning an Afghan hound in 2005. "Dr. Hwang's past is obviously controversial, but we feel that his lab and his record when it comes to dog cloning are the best in the field," Mr. Hawthorne said. "He's been very open with me about admitting his mistakes. Nobody says he lied about cloning animals." Elizabeth Wictum, associate director of the Veterinary Genetics Laboratory at the University of California, Davis, said that earlier this year, she and her staff had taken sets of DNA extracts from Mr. Hawthorne's puppies and compared them with stored samples of Missy's DNA, and concluded that the results were "consistent with clones." "The puppies had the same nuclear DNA as Missy, and different mitochondrial DNA, which is what you get from a cloned animal," Ms. Wictum said. "If somebody were trying to, say, sneak in two samples from the same dog or an identical twin and claim that one was a clone's, there would be no differentiation between the nuclear and mitochondrial DNA." Missy 1.0 - Mira and MissyToo's "genetic donor," as Mr. Hawthorne calls her when he's speaking technically - was his mother's dog. To date, he said, there are four Missy clones running around, all born between December 2007 and June 2008. Mira lives at Mr. Hawthorne's house in Mill Valley and MissyToo between homes in Mill Valley and San Francisco, both owned by his mother's boyfriend; clones No. 3 and 4 were given away to friends and now live in Phoenix and Boulder, respectively. (Two others were produced but died of parvovirus as newborns.) In July, Mr. Hawthorne held a series of online auctions, offering his services to four high bidders who agreed to pay from $130,000 to $170,000 for clones of their dogs. (To offset accusations of elitism, Mr. Hawthorne also held a search for a fifth, pro bono client.) Over the last few weeks, the first three puppies from this group were born in South Korea. Mr. Hawthorne claims Missy's clones are the world's first commercially cloned dogs, although RNL Bio, a Korean company with which he is embroiled in a legal dispute over patent rights - and the chance to dominate the dog-cloning market - has also made this claim. Between 2004 and 2006, another company Mr. Hawthorne ran cloned cats for a handful of paying customers. Dogs are among the most difficult mammals to clone, scientists say, because their reproductive systems are highly atypical, but Mr. Hawthorne thinks that the market is keen. When Mr. Hawthorne recalls Missy, he tends to wax eugenic. "She was an amazing dog: superior intellect, incredibly beautiful, obedient, a phenomenal temperament," he said. "I especially loved her majestic plume of a tail." And in the clones, as he put it matter-of-factly, "all those qualities are represented." As for some of the discrepancies, the clones vary in size and color, Mr. Hawthorne said, primarily because they were born months apart, and none are fully grown yet. "The dark part of their fur starts out reddish-black and gets blacker over time," he said. "Except on the faces, which start out black and go white within the first year." Each clone's embryo was created by joining Missy's nuclear DNA with the enucleated (which is to say, DNA-stripped) egg of a different dog. The eggs, with their new DNA from Missy, began to grow and divide. After that, each was carried to term in the uterus of still another dog - a step which, though it has no bearing on the dogs' genetic makeup, can affect such external traits as the waviness of fur and the up-or-down pointing of the ears. (It has to do with different levels of collagen.) On his afternoon trek around Lake Lagunitas, Mr. Hawthorne did his best to explain these matters to his audience, picnicking families who sat transfixed as the two dogs he calls "clisters" - his made-up word for clone sisters - took turns chasing each other off leash. "Can they clone coconut?" a girl asked. "Sorry, we don't do vegetables," Mr. Hawthorne said. A parent clarified: Coconut was the family dog's name. "Can you clone a three-year-old housebroken dog?" Mr. Hawthorne went on gamely. "Learned behavior, no. But a lot of behavior is hereditary," he said. While he does acknowledge that when it comes to such highly trainable creatures as dogs, it's pretty difficult to know where nature ends and nurture begins, he said that in the case of his dogs, the ambiguities have nothing on the essential Missy-ness of the clones. "The girls love to run after each other," he said pointing at the dogs in the distance. "You see the speed and athleticism? That's part of what made me want to do this. There are dogs that are faster on a straightaway, but I'd never seen a dog make turns like this until Missy." Mr. Hawthorne sees himself as a cultivator of prodigious talent - from the clones to his team of scientists in South Korea to his 8-year-old son, Skye, who had accompanied him and the dogs on the hike. Skye is in third grade but is already studying high school algebra. Mr. Hawthorne brought along a notebook with a handful of quadratic-equation problems, in case his son got restless. Last spring, Skye completed a science project, "Cloning Grandma's Dog" that included a behavioral comparison chart. Among other findings, the study concluded that Mira shares Missy's fondness for broccoli and "lots of snuggles" - both dogs scored five out of five points in these categories, in addition to the one for "likes long walks." ("Most dogs do," Skye noted under "comments.") Two key matters of variance were "Jumps into cars" ("Clone still learning which car is ours") and "Hates camera flash" ("Clone did not respond to standard flash"). Ultimately, Skye determined that Mira looked a lot like Missy but that their behavior was only 77 percent similar. "But that was April," Mr. Hawthorne said. "I think they're a lot more similar now." LIVING with a clone, Mr. Hawthorne claims, is a lot like living with the original dog. "It's totally as if I've got Missy in my house, once you get over the 'wow' factor," he said. He and Mira and Skye inhabit a two-story "1950s futurist house" built into a hill in Mill Valley (Mr. Hawthorne is divorced and shares custody of his son with his ex-wife - "an excellent genetic donor, by the way," he said of her). At night, he said, Mira "puts Skye to bed," which means she walks with him to his room and ascends the stairs of his loft bed with him, waiting to be told "Good night" before she leaves. Mira is an outdoor dog, as Missy was, and sleeps on the front steps. And who says goodnight to Mira's fellow clone, MissyToo? Mr. Hawthorne gave her to his mother, Joan Hawthorne, who still misses the original Missy. But she has yet to take a liking to Missy's progeny, and the dog has lived primarily with paid "handlers" in the Mill Valley pied-à-terre of her longtime companion, John Sperling. "They're not at all alike," Ms. Hawthorne said of the old Missy and the new one. "In looks, they are a little bit, of course. But, I mean, the puppy is delicate and aggressive. Missy was robust and completely calm." She added, "Missy wouldn't come through my home and knock over every wineglass." Besides, she adopted another puppy not long after Missy died. "I already have a dog - a real dog." The idea of trying to clone Missy in the first place came from Mr. Sperling, who has been in the family picture since Mr. Hawthorne was 4 years old - Mr. Hawthorne typically refers to him as his stepfather - back in 1997. At the time, Mr. Hawthorne was at a sort of career crossroads, having worked in the interactive video-production business and just returned from five months riding a motorcycle across India. Mr. Sperling, who would not agree to be interviewed, is a billionaire who founded the University of Phoenix, the adult-education institution with nearly 200 locations. Mr. Hawthorne said that his various fruitless attempts at cloning were paid for by more than $15 million of Mr. Sperling's money, some of which went to purchasing several competing and related biotech businesses in what Mr. Hawthorne described as an industry roll-up. "It's kind of weird," Mr. Hawthorne said. "They spent 10 years waiting for this to happen and then they don't even want the dogs living with them." In recent weeks, he added, Mr. Sperling has been making an effort to keep MissyToo with him at his house in San Francisco. ("He's going to have to launch on some serious dog training," Ms. Hawthorne said of this new development.) On the day of the hike around the reservoir, Mr. Hawthorne, with a reporter as his audience, telephoned Nina and Ed Otto, auction winners in July - they paid $155,000 - who were about to learn that they were the proud owners of the project's first spawn. A clone of Lancelot, their late yellow Labrador, had been born a couple of days earlier in South Korea. From the courtyard of a Starbucks, he dialed their home in Florida on his cellphone. "Your little Lancelot is here," Mr. Hawthorne informed Mrs. Otto. He followed up with date and time of birth, weight (540 grams, or 19 ounces), then e-mailed two pictures of the newborn as they spoke. "Breathe, breathe," Mr. Hawthorne said. "How does he look?" "He looks like Neanderthal Man," Mr. Otto called out from the background. "No, he doesn't," his wife said. "I can't say he looks exactly like our other one," Mr. Otto shouted. "Yes, he does," his wife said. Several days later - the Ottos will not see their new dog face-to-face until he is 10 weeks old, when they'll have him flown to the United States - Mrs. Otto said they were ecstatic about "having the essence of our Lancelot back." "We can barely contain ourselves," said Mr. Otto, the chief executive of a medical-equipment company whose father was an early Nascar promoter. "We have 8 children and 11 grandchildren and 9 dogs. Lancelot was the dog that was most humanlike in his behavior. He died last year - cancer." Mrs. Otto said: "The only problem with dogs is that they have such a short life." "Cloning means you could have the opportunity to have the same dog with you for your entire life," Mr. Otto said.