Breeding Plans

Cloning


Stubborn As a Mule


They said it couldn't be done. But one scientist was stubborn as a mule -- and then he cloned one.


Gordon Woods, a researcher at the University of Idaho in Moscow led a team that has cloned the first equine, a mule they've named Idaho Gem. He was born on May 4. Researchers expect another two more clones will be born this summer.


Mules, a cross between donkeys and horses, are almost always incapable of reproducing on their own because they are sterile.


Most cloning researchers thought it was impossible to clone any equine because of certain peculiarities in their genetic makeup. But Woods was determined to make it work.


"Anybody who wanted to come into my lab to clone a horse, (let alone an animal that can't reproduce), had to be short a few marbles," said Kenneth White, a researcher at the University of Iowa who participated in the study. "But I'm obviously not very persuasive because I tried everything I could to discourage this idea, and he obviously didn't listen very well."


Idaho Gem is Woods' baby, so to speak, and he spoke like a proud father at a press conference on Thursday. "Idahoans are solid, they're fiercely independent and they're persistent," Woods said as the mule was revealed to reporters on the Idaho campus. "In honor of that may I present to you, Idaho Gem."


The successful pregnancies came after 307 tries, so the Idaho method is still inefficient. But researchers consider the healthy birth of a cloned equine a major feat.


The researchers created Idaho Gem using a cell from a mule fetus that was created and frozen in 1998. They implanted the fetal cell into an egg taken from a mare, after removing the nucleus of the egg.


Normally, the presence of sperm triggers cell division when an egg is fertilized. Since cloning doesn’t involve any sperm, the researchers used a calcium mixture to trick the cells into dividing. They then implanted the cell into a surrogate mare. Their ultimate goal is to clone a horse. Researchers believe their technique will work equally well in horses.


Idahoan businessman and president of the American Mule Racing Association, Donald Jacklin, funded the project with a $400,000 donation. He wanted the researchers to create a clone that would be directly related to his champion mule, Taz.


Since mules are almost always sterile, mule-racing enthusiasts can't breed the animals along family lines. So cloning is of particular interest to them.


The University of Idaho owns the cloned mule, but Jacklin said he hopes they'll allow him to race Idaho Gem, or one of the other clones, when they turn three.


"He sure sounds and looks like he's going to be a Taz gene-type," Jacklin said. "He's the property of the University of Idaho, but I definitely have a strong interest in securing his availability through a lease or whatever method to take on one of the babies for racing."


Idaho Gem is the most recent in a series of cloned mammals that began with Dolly the sheep cloned in 1996 at the Roslin Institute in Edinburgh, Scotland. Since then researchers have also cloned cats, cows and pigs. Dolly was euthanized in February because she was suffering from a lung infection, and she had shown signs of arthritis. Some researchers believed she might have been aging more rapidly than she should.


But the Idaho researchers who cloned the mule said they weren't concerned that Idaho Gem would have aging problems. "Clinically we saw no evidence of any placental or fetal (abnormalities)," said Dirk Vanderwall, another Idaho researcher who participated in the study.


The researchers bred Taz's parents -- a jack donkey and a horse mare -- to create an embryo in 1998. They let it develop for 45 days, then froze the cells in anticipation of using them in the cloning experiment. It took four years to solve the problems implicit in cloning equines.


The key to the problem, they believe, may have been calcium. A spike in calcium occurs after conception and induces the division of cells, which eventually leads to an embryo. By experimenting with different levels of calcium, they achieved 21 pregnancies, and it appears the mares will carry a total of three to term. The researchers expect the other two to be born in June and August.


The Idaho researchers surgically implanted the early embryo into the mare's oviduct -- the tube that passes eggs from the ovary and where fertilization typically occurs. This also may have increased their chances of a live birth.


A very young embryo can't survive in the uterus, only in the oviduct. Other researchers are working on ways to grow horse embryos in a dish until it's able to survive in the uterus. But an oviduct is a much more friendly environment for a blastocyst (young embryo) to develop.


"That's the best place for an embryo," said Katrin Hinrichs, a professor in veterinary science at the Texas A&M University. "You cannot improve on mother nature."


Surgery is expensive, however, as well as inefficient. So, without the generous funding received by the Idaho researchers, Hinrichs and her colleagues have impregnated one horse with a clone by growing the embryo in a dish first, then implanting it into the uterus.


"We're trying to do it in the most cost-efficient manner," she said. "We only had to transfer five blastocysts to get the one pregnancy we got."


Mule race soon to see double


Project Idaho clones to begin training for track competition


By MIKE McLEAN


Coeur d'Alene Press


POST FALLS - Two mule clones are following the tracks of their natural-born brother. University of Idaho mule clones Idaho Gem and Idaho Star are off to the races.


The university leased the clones to businessmen Don Jacklin of Post Falls and Roger Downey of Albuquerque, N.M. They serve as president and vice president, respectively, of the American Mule Racing Association. ?The two guys are doing marvelous,? Jacklin said of the mule clones.


Jacklin, owner of champion racing mule Taz, was one of the biggest sponsors behind Project Idaho, the UI program that was the first to successfully clone a member of the horse family.


The genetic duplicates are full brothers of Taz, a champion racing mule. The clone DNA comes from skin cells of the same fetal mule.


Racing will test the clones' genes and athletic ability. Jacklin thinks they have the potential to be champions like Taz. "I was just so impressed by them, their attitudes, the width of their chests, their temperament, and, of course, their genetics."


Jacklin leased Idaho Gem, because he was the first of three mule clones. A mule is a hybrid, born of a donkey father and horse mother.


With rare exceptions, mules are sterile and can't reproduce naturally.


Idaho Gem was born May 4, 2003, Utah Pioneer was born on June 9, 2003, and Idaho Star debuted on July 27, 2003. Gem has been sent to a trainer in California. Downey is having Idaho Star trained in Montana.


It's still a long way to the track. The pair must complete nearly a year of training before they reach a racetrack. "They will have light training until January or February of 2006," Jacklin said. "Then we'll move to race training."


That's when the two clones will begin to prove their fitness to race and address the debate about whether genetics or environment makes for winners or also-rans. "We'll be able to evaluate different management techniques," Jacklin said. "We'll have a good feeling of the different environmental factors on who trains who ... and how they are brought forward into the racing circles."


Their genetics are showing through. "If you put them all separate from one another and look at one and then walk over and look at the other, you can tell they are absolutely physically identical," he said. "But if you put them together, you can see a little height and size differential."


There's about an inch difference between the Gem and Star. "There are a few more pounds on the first-born," he said. Although they are genetically identical, they each have their own personality, based on their surrogate mares to which they aren't genetically related.


"They are spinoffs from their mothers," Jacklin said. "If their mother acts one way, they kind of mimic the action." Idaho Star was the last clone born. His mother was a "nasty herd boss," who pushed others around and let them know she was in charge, Jacklin said.


"The baby would try to act like that even though he was the smallest of the group," he said. "He would try to dominate the other two brothers when they were together." Yet, Star seems to be the most willing to accept training. "He seems to understand it quicker and wants to please quicker," Jacklin said.


Idaho Gem is easier to be around, but his handler in California said he's not as easy to train. "He's a little more reluctant to accept it," Jacklin said.


The British television company, Discovery, approached Jacklin and Downey to do a documentary on the clones. "I think they want to do it on their racing performance for comparison of clones," Jacklin said. "We're excited about working with them."


Taz is home recovering from a tendon injury he suffered in a race in Pamona, Calif., last year. Jacklin is giving the 11-year-old champion a full year to recuperate. "I just want to make sure he's totally healed, so he can race again," Jacklin said.


Jacklin owns four other racing mules. Three of them are related to Taz. He also is a one-third partner in another mule.


The mule racing season began last week in Winnemucca, Nev. The opener is followed by a race in Oregon. The remainder of the races are on California tracks, extending into October.


Unlike thoroughbred horse racing, mule racing has no restrictions on the use of artificially bred animals. Utah Pioneer, the middle clone, remains at the UI campus. The clone is not alone, said Dirk Vanderwall, UI assistant professor of equine reproduction and a veterinarian.


"We've already had several school groups come and see him since Gem and Star left," Vanderwall said. The added human attention and the lack of distractions from his stable mates also led Utah Pioneer to become more focused on people. He had been the most reticent of the clones to seek attention.


Idaho Gem and Idaho Star were transported to trainers in 2005 to prepare them for racing in 2006. Idaho Gem and Idaho Star both won their first races on June 3, 2006, separate trial races for the Humboldt Futurity during the Winnemucca, Nev., Mule Races, Show and Draft Horse Challenge, June 3 and 4.


In the June 4 futurity, Idaho Gem finished third and Idaho Star finished seventh. Idaho Gem won his next race at the San Joaquin Fair in Stockton, June 21. His time of 20.724 seconds over the 350-yard course was the fastest time by a 3-year-old mule through the end of July, the halfway point in the mule racing season. Idaho Gem also collected two seconds in photo finishes with the racing mule Out of My League. The total margin of victory between the two mules in the two races was .043 seconds. Through his first six races, Idaho Gem collected two firsts, two seconds a third and a fourth.


On June 4, 2006, Idaho Gem finished 3rd in the Winnemucca Mule Race. This was the 1st showdown between cloned and natural-born mules.


August 8, 2003; Since the birth of Dolly the Sheep, cloning scientists have so far cloned sheep, cows, pigs, goats, cats, and now following the birth of the mule Idaho Gem on May 30th below, an Italian Team (The Laboratory of Reproductive Technologies in Cremona) is the first to clone a horse, as reported Thursday in the journal Nature.


Born naturally on May 28th, the female foal's name is Prometea named for the Greek God Prometheus (a Titan who was punished for stealing fire from Mt. Olympus). (In a twist for the growing number of cloned species, the Haflinger mare that gave birth to Prometea was also the source of her DNA, meaning she and her foal are identical twins.) 800 enucleated eggs, obtained from ovaries gotten from slaughterhouses, were inserted with DNA from skin cells taken form either a male or a female horse. 22 embryos successfully developed to the seven-day stage of development. 17 were placed in mares, and 4 implantations resulted. Three ended in miscarriages, and finally one foal was born in tact. This is not a great yield, speaking commercially, but at least it's an existence proof. Maybe the canine cloners at Texas A&M will pick up a few tips after many years of failure. They are awaiting the birth of a cloned American quarter horse, but were just beaten by the Italian team.


April 28, 2005; Born on March 13th, Paris Texas, the first cloned foal in the US was announced by Texas A&M University researchers at College Park, TX [1]. (A female horse was previously cloned in Italy.) It took over 400 attempts over a six-month period. Six embryos were created, and one of them was successfully brought to term in a surrogate mare named Greta. The pregnancy took 12 1/2 months. (The normal gestation time for horses is 11 months.) Texas A&M had previously cloned pigs, a Boer goat, an Angus bull, a Brahma bull, and a deer.


This month, a French-Italian collaboration announced the successful birth of a foal cloned from a gelding. Now two months old, the foal was produced by the French genetic engineering company Cryozootech and the Italian reproductive technology lab LTR-CIZ. The lab's team, headed by Cesare Galli, has improved on techniques it used two years ago to produce the first horse clone, a mare. From 200 nuclear transfers using skin cells from Pieraz, a retired thoroughbred Arabian endurance champion, the researchers got 34 embryos and three pregnancies, one of them successful. [2]


Galli has predicted that cloning will revolutionize the horse-racing industry. But at present, the thoroughbred racing community doesn't even permit artificial insemination, much less cloning. Paul Struthers of Britain's Jockey Club says racers have a very restricted gene pool and "there would be very serious implications for the long-term welfare of the thoroughbred were the gene pool to be reduced further" by breeders all going after the progeny of superachievers. But cloning could have a future with horses intended for show jumping, dressage, or endurance racing events with fewer breeding restrictions. "Over 90 percent of dressage stallions are gelded to make them more manageable," [So, that's why they do it. I could never figure out why anybody would ever want to geld a race horse just by common knowledge from reading the sports pages, since, obviously, I'm not an insider in this particular sandbox.] says Nicolas Robin of Cryozootech: "So imagine how many gene lines are lost." But no longer. The company is preserving cells from some 30 prize stallions and plans to market semen from their clones.


1. AP, "Horse Cloned by U.S., French Research Team: Believed To Be a First in North America -- the Foal Named Paris Texas -- Makes His Debut," The Los Angeles Times, p. A23 (April 28, 2005). 2. Constance Holden, "Champion Racer Cloned," Science, Vol 308, No. 5722, p. 628 (April 29, 2005).

March 31, 2006; ViaGen of Austin, TX and Encore Genetics, Ltd. of Weatherford, TX announced that foals, Royal Blue Boon Too and Tap O'Lena are the first commercially cloned (cutting) horses. The price for creating a clone of a horse they say will be set at $150,000 for the first and $90,000 for the second clone of the same animal. On March 9th, a clone of the mare Tap O Lena and another clone was born to a filly at the same farm. "Two more clones of the mare Bet Yer Blue Boons are expected to be born any day," said Executives for ViaGen and Encore Genetics.


As distinguished from The National Cutting Horse Association, which trains horses to help rope calves from a larger herd, The Jockey Club, which organizes thoroughbred racehorses, has strict prohibitions against cloning, as does The American Quarter Horse Association. Curiously, The Humane Society of the United States issued a statement Thursday criticizing these two Texas companies for subjecting horses to commercial exploitation, saying that it has "no legitimate social value." "The high failure rate in cloning and the high rate of congenital birth defects and other illness are reasons to avoid this technique."


ViaGen is controlled by Dr. John Sperling of Phoenix, AZ, an octogenarian billionaire. He is also the owner of Genetic Savings and Clone, Inc. that has been working on cloning dogs for some years and is expected to make an announcement some time later this year.


Refs.: 1. Andrew Pollack, "Goodbye Dolly: Up from Sheep to Cloned Horses," The New York Times, p. C4 (March 31, 2006). 2. Andrew Pollack, "Latest Clone Line: Champion Horses," The International Herald Tribune (April 1, 2006).