"Ping'an" was born, and the clone cat in colleges and universities welcomed the new era
The first cloned cat completed by a university in China was born in Qingdao Agricultural University and became an "Internet celebrity" on the Internet
In November this year, the mammalian somatic cell cloning base of Qingdao Agricultural University ushered in a special "newborn" - a lively and cute orange cat. This little orange cat is different from our common cats, it is the first somatic cell cloned cat in China that uses domestic equipment, reagents and consumables in the whole process, which means that China officially has a complete industrial chain in the field of animal cloning. The national and global cloning teams have set their sights on this soft and cute kitten, and its birth marks the "20 version" appeared, and when it comes to cloned cats from Qingdao Agricultural University, we have to start from the "1Version 0 "- Ping" said.
Clone cat "Heian" (right). On December 24, 2020, in the laboratory of Qingdao Agricultural University, a black and white kitten was sleeping soundly in the incubator, looking healthy and cute. After a series of physical examinations, the 75-gram kitten was named "Ping'an" by the researchers, hoping that its research results as a cloning technology can survive safely and healthily, and at the same time, it is hoped that it will be born at the end of the year to bring peace and good luck to the new year.
A cloned cat was born in a university, which instantly attracted attention in China. Where does "peace" come from?Dr. Zhao Minghui, the host of the project and the College of Life Sciences of Qingdao Agricultural University, introduced: "The body of this cloned cat is an adult female forest cat crossed by a wild forest cat and a Chinese pastoral cat. Researchers isolated and cultured fibroblasts from the cat tissue, and used the eggs of Chinese pastoral cats as nuclear receptors to produce multiple somatic clone embryos through nuclear replacement and artificial activation. After the cloned embryo was transferred to the recipient female cat, after 62 days of gestation, a forest cat somatic cell cloned offspring was successfully obtained. After the appraisal of the Beijing Zhongzheng Judicial Appraisal Institute, it was determined that the forest cat was a cloned individual. ”
62 days of gestation requires a lot of work before and after. Recalling the process of cloning a cat from scratch, Zhao Minghui introduced: "To put it simply, the cloning technology of a cat requires first to get somatic cells containing local genetic material from the cloned body, and then obtain the oocytes of the conceived cat, which contain cytoplasm and nucleus. On this basis, we need to remove the nucleus from the oocyte and replace it with the somatic cell of the body, so that the genetic material and genetic information of the body are introduced into the egg. After introduction, it is artificially activated to make it a cloned embryo similar to a fertilized egg, which is then placed in the oviduct of the female cat for normal development, and the cloned cat is obtained through breeding. The last link is the care and breeding of cloned cats, and the general cloned animals are such a process. ”
The research project on cloning cats at Qingdao Agricultural University was prepared in 2019 and implemented in July 2020. "In 2016, I independently tackled the development of somatic cell cloning dog technology, and I hoped to make a new breakthrough, so I chose to clone cats. Zhao Minghui said that the body of the cloned cat "Ping'an" is an adult female forest cat crossed by a wild forest cat and a Chinese pastoral cat. At that time, in the animal hospital, the owner of this cat took it to be neutered, and we found that this cat has a flat nose bridge, a relatively large face, and a long-haired cat, which is a good choice as a cloning object. We obtained the consent of the cat owner, took a little ** from this cat, isolated some fibroblasts from it, and obtained a large number of cells through sub-amplification for cryopreservation. ”
Recalling the difficulties encountered throughout the project, Dr. Zhao Minghui said: "The first bottleneck encountered in the experiment was the in vitro maturation of cat eggs. "Since the cat eggs come from the ovaries that were cut during sterilization in the pet hospital, the cat eggs collected on these ovaries cannot be used directly and must go through a period of in vitro maturation. For the in vitro maturation of cat eggs, Zhao Minghui and his team members conducted a lot of experiments, and finally overcame this problem by adding appropriate amounts of hormones and growth factors. "The birth of 'Ping'an' also proves that we have succeeded in the in vitro maturation of cat eggs. Zhao Minghui said.
About me: In September, I received higher education at birth, engaged in volunteer work such as embryo engineering and somatic cell cloning, and lived in a large villa at Agricultural University. i can speak english。"About you: British longhair, silver gradient, green eyeballs, pure London accent first, disease-free and moss-free. Together, we can study the topic of breeding cloned cats. On August 27, 2021, Zhao Minghui posted a "cat high-quality women" courtship information on WeChat Moments on the Internet, and "peace" once again appeared in the public eye. "In October, 'Ping'an' came into heat, which is a sign of sexual maturity, indicating that 'Ping'an' reproductive function is normal. Since there were no male cats in the laboratory, I wanted to find a cat for 'Ping'an' to match, but I didn't expect to be discovered by reporters, and things fermented. ”
After the information was released in the circle of friends, many enthusiastic netizens hurriedly posted their cats' ** "application", and the comment area once became a cat drying area. Zhao Minghui said that there are more than 20 cats that have come to the 'blind date', and some of them are very good, and they have also helped the candidate cats to be tested for genetic diseases and infectious diseases. On December 24, 2021, the team members of the clone cat team of Qingdao Agricultural University found a suitable breeding partner for "Ping'an", and "officially announced" that it would get off the list on its birthday. Team members introduced: "The other party's name is 'Hanamaki', and he is also a long-haired cat, healthy, good-looking, gentle and has passed the checks. ”
The story of the success of the cloned cat "blind date" spread all over the Internet and amused many netizens. For Zhao Minghui, on the one hand, he hopes to be a mother "safely", and on the other hand, he wants to verify that his fertility is normal. Zhao Minghui believes that strictly speaking, it cannot be said that the cloning of cats is successful, because the fertility has not yet been verified. "There are three important evaluation criteria for cloning animals, one is the absence of defects in appearance and bodily functions, the second is longevity, and the third is fertility. Fertility is a very important indicator. Zhao Minghui said.
What does it mean to be born with a cloned cat?The birth of "peace" has brought everyone deep thinking. In this regard, Zhao Minghui believes that the establishment of cat somatic cell cloning technology is not only conducive to promoting the development of animal disease models and drugs, but also plays an important role in the field of biomedicine, the pet industry and the protection of endangered species.
When it comes to medical research, cats are important laboratory animals, disease model animals, and companion animals. "According to the statistics of the OMIA database, 361 genetic diseases have been found in felines, of which 227 are expected to develop animal models of similar diseases, and the pathogenesis of feline epilepsy, hyperlipidemia, chondroplasia and some tumors, AIDS and other diseases is particularly similar to that of humans. What has attracted more attention from researchers is that cats have ACE2 receptors that are highly similar to human structures, and this receptor is the main receptor for SARS and the new crown pneumonia virus to infect the human body, which makes cats have the potential to be infected with SARS and new crown pneumonia. Using cat cloning technology and gene editing technology, individuals carrying specific diseases can be mass-produced for use in medical research and drug development.
In terms of pet market development, the data of China's pet industry in 2019 shows that the domestic pet (cat, dog) market size has reached 202.4 billion yuan in 2019, a year-on-year increase of 185%。Among them, the size of the pet cat consumer market was 78 billion yuan, a year-on-year increase of 196%, exceeding the growth rate of the dog consumer market. Although the current market for cat somatic cell cloning is between 15 and 250,000 yuan, there are still more and more pet owners who choose to "reborn" their pets through somatic cell cloning after their pets are sick and cannot be treated or pass away. In addition, the early embryo gene editing technology derived from cat somatic cell cloning also brings hope for pet genetic diseases.
In addition, according to Zhao Minghui, cat somatic cell cloning also plays an important role in the preservation of endangered species. Felines are an important part of the earth's animal resources. With the exception of domestic cats, all other cats are endangered or threatened. According to data released by the International Union for Conservation of Nature, three of China's 21 wild cat species are endangered, two are critically endangered, and seven are vulnerable or near threatened. When an animal loses its fertility in old age, it means the functional extinction of the species. The use of domestic cat eggs for xenocellular cloning is the only technical means that can reproduce endangered animals in large quantities.
The whole process of domestic cloning cat technology is upgraded
Twenty years of technological upgrading, breakthrough in the "neck" technology of imported drugs, and the birth of "nationally produced" cloned cats
Three years after the birth of the cloned cat "Ping'an", there was good news from the mammalian somatic cell cloning base of Qingdao Agricultural University - a lively and cute little orange cat was born. Different from "Ping'an" three years ago, this little orange cat is currently the first somatic cell cloning cat in China that uses domestic equipment, reagents and consumables in the whole process, marking a new breakthrough in the field of animal cloning technology in China. Yuan Bao, vice chairman of the Animal Reproduction Branch of the Chinese Association of Animal Husbandry and Veterinary Medicine and professor of the College of Animal Science of Jilin University, said that this achievement has fundamentally changed the situation of using imported products for animal cloning process equipment and reagents, and provided the possibility for reducing the cost of animal cloning. The birth of this "purely domestic" cloned cat is a key breakthrough in the localization of the whole process of cloning animals in Qingdao Agricultural University.
Qingdao Agricultural University "pure domestic" cloned cats. Following the birth of "Heian", on November 19 this year, another cloned cat with which it was not related by blood was born. "Ping'an" is small and skinny, and the newly cloned wild orange cat is a "big fat boy" who weighs 141 grams. If "Ping'an" is the first healthy and surviving clone cat in a domestic university, then this newborn orange cat is even more remarkable, it has become the first full-process localization of the clone cat in China.
According to Zhao Minghui, a doctor from the College of Life Sciences of Qingdao Agricultural University, this cloned orange cat is the crystallization of the cooperation between Qingdao Agricultural University and Qingdao Panji Science and Technology, and it is a commercial clone. "The breakthrough of this technology has greatly reduced the cost of cloning consumables and shortened the road to cloning industrialization. Zhao Minghui introduced that as early as 2020, the team successfully obtained the cloned descendants of the somatic cells of the British long-haired cat through tackling key problems, but at that time, most of the reagents and core equipment used to clone "Ping An" were "made in foreign countries". Later, when Zhao Minghui's team was cloning pig somatic cells, many embryos died "en masse", which made them try to use domestic drugs.
In the past, the reagents used in the development of cloning technology and the two most important large-scale instruments, the micromanipulator and the electrofusion instrument, had to be imported from abroad, which was high and the channel was unstable. Zhao Minghui said that the team's determination to do the localization of the cloning industry chain stemmed from a failed experiment. When they cloned pigs, there were quality problems with the foreign reagents they bought. Originally, I had the mentality of trying it out, but I didn't expect that the domestic drugs worked very well, so Zhao Minghui's team made a bold attempt - simply replace all the drugs and equipment with domestic ones. In the meantime, because some drugs could not be replaced locally, they changed the formula, optimized the fixture and fusion instrument of the micromanipulator, and finally realized the localization of the whole process of cloning.
From October 25th to November 2nd, Zhao Minghui's team cloned the first batch of 20 somatic male Licha black pigs with the whole process of localized equipment and drugs. On November 19, the whole process of localized somatic cell cloning of the little orange cat was also born.
Clone the Black Pig. On the road of scientific research of the "national" cloning technology, Zhao Minghui's team found a way to clone the black pig without using imported instruments and reagents from the project of cloning the black pig.
In China, the industrialization of cloned pigs has been quite mature. In the past few decades, due to the influence of marketization, people have long pursued the high lean meat rate and high growth rate of commercial pig breeds, ignoring the protection and utilization of germplasm resources of Licha black pigs with excellent traits, resulting in a sharp decline in the number of Licha black pigs. Zhao Minghui introduced that the pig seed industry innovation team of the College of Life Sciences of Qingdao Agricultural University cooperated with Jiaozhou Maohua Licha Black Pig Breeding Farm, Qingdao Panji Science and Technology and other units to carry out relevant research work on the protection and innovative utilization of Licha black pig germplasm resources.
A total of 1093 cloned embryos were obtained and transferred to 5 ** sows, with a pregnancy rate of 100%. 5 ** sows successfully gave birth to 20 male Licha black pigs from October 25 to November 2. Zhao Minghui said. On November 9, the pig seed industry innovation team of the College of Life Sciences of Qingdao Agricultural University successfully completed the cloning of China's excellent local pig breed Licha black pig, and realized the localization of equipment and reagents required for the whole process of the work.
Compared with pigs, somatic cell cloning in cats requires higher equipment, reagents, and consumables, and is more delicate to operate. In the previous animal cloning, most of the reagents and core equipment were purchased from abroad, which was subject to technical limitations.
The cloned cat is an animal born from the somatic cell cloning operation, without a father, and the mother is just a mother, the genetic material is exactly the same as the body, but its lifespan after birth is calculated from 0, and there will be no premature aging and the like. Zhao Minghui introduced that it is no different from the appearance of the female cat of the body, and the difference is in the different life experience. Talking about the birth of this "purely domestic" cloned cat, Zhao Minghui introduced, "We have screened some domestic easy-to-use reagents, and then improved the reagents that are not available in China." In June this year, Zhao Minghui's team basically completed the localization and substitution of reagents. At the same time, the team inspected and searched for manufacturers of micromanipulators and fusion instruments for cloning core equipment in China for many times, and finally established contact with a domestic instrument company.
Zhao Minghui told reporters, "The birth of this cloned orange cat shows that there is no significant difference in the performance of our domestic reagents and foreign reagents, and it can even be said that they have reached the advanced level." Through this project, we found that the domestic equipment is better controlled than foreign countries in terms of details, indicating that China's manufacturing of high-end equipment in the field of cloning has also reached the world's advanced level. ”
The successful birth of this cloned cat is not only a major breakthrough in the field of animal cloning technology in China, but also a microcosm of the rapid development of China's high-end equipment manufacturing, precision machining and chemical industry, which proves that China has a complete industrial chain in the field of animal cloning.
In the field of biology, animal somatic cell cloning refers to the asexual reproduction of organisms through somatic cell nuclear transfer, as well as the offspring individuals with the same genotype formed by asexual reproduction, and the research of cloning technology is of great significance in the field of biomedicine, pet industry and endangered species protection. In 1996, the birth of Dolly the cloned sheep caused a sensation around the world. Today, cloning is no longer a secret. In China, cloned fish, cloned mice, cloned rabbits, etc. have been introduced, cloning technology has been updated and iterative, and in Qingdao Agricultural University, with the efforts of a group of scientific researchers, in the past 20 years, cats, pigs have been successfully cloned.
In 2001, China's first and second healthy living cell cloned cattle "Kangkang" and "Shuangshuang" were born in Qingdao Agricultural University. In 2019, a joint research team of Shandong Academy of Agricultural Sciences and Qingdao Agricultural University successfully cloned a long-haired somatic cell clone dog "Lulu". In 2020, Zhao Minghui successfully obtained feline somatic cell cloning offspring, making Qingdao Agricultural University the first university in China to successfully complete feline somatic cell cloning. In 2023, 20 male Licha black pigs cloned by national production equipment and reagents will be born in Qingdao.
After more than 20 years of research, the cloning technology of Qingdao Agricultural University has gradually stabilized and efficient, the probability of success is controllable, and the instruments and reagents are no longer dependent on imports. For a long time, Qingdao Agricultural University has been carrying out basic and applied research for production practice. In the next step, Zhao's team will continue to focus on gene editing in cats.
The post-85 team will carry out the "cat" business to the end
Dr. Zhao Minghui's team has improved its independent innovation ability and made major breakthroughs in cloning cat technology
Young, shy, and mingling with team members is the first impression of Dr. Zhao Minghui by Qingdao Morning Post reporters. When he put on his lab coat and sat in front of the microscope, this "cat expert" had a serious and rigorous attitude. From 3 years ago to the present, in Zhao Minghui's view, the birth of cloned cats not only represents a major breakthrough in science and technology, but also witnesses the growth of a hard-core "clone team" of Qingdao Agricultural University.
Zhao Minghui, Ph.D., College of Life Sciences, Qingdao Agricultural University. Back then, "Ping An" became popular all over the Internet, and in the laboratory of Qingdao Agricultural University, it also became a "group pet" because of its special identity. Not only is the place where you live cultural, but you are also very particular about your food. "Afraid that 'Ping'an would get sick, we let him live in an air-conditioned house with purified air, bought the best cat food, gave him mineral water, and usually brought cooked chicken breasts for him to eat while it was hot. Zhao Minghui introduced, when he was born, "Ping'an" was at the lower value of 75 grams of normal cat weight, and when he was 4 months old, the weight was "off the table", and now seven or eight pounds is normal. "Don't look at 'Ping'an' with big hair, it's actually not much different from ordinary cats, both docile and honest. Zhao Minghui introduced that if you don't take the initiative to touch it during the day, it will stick to your leg and rub left and right in a circle, which is a unique trick of "peace" to hug.
When the weather is good, "Ping'an" will be carried to bask in the sun, and only after jumping up and down and playing with his favorite paper ball will he reluctantly return to the nest. Although "Ping'an" is a clone of Zhao Minghui's team, what few people know is that Zhao Minghui himself is severely allergic to cat hair. "I have medicine at home, in my car and in all the offices, and I take it whenever I feel like I have a reaction. Even so, Zhao Minghui was still willing to be a "cat father". In order to clone the cat "safe", Zhao Minghui also prepared a pink nest for it, and often carried it around the campus.
"Ping An" has had its own traffic since birth, and behind this attention is the doubling of Zhao Minghui's workload. "Originally, it was okay to concentrate on one thing in the laboratory, but after the birth of 'Ping An', there were many companies that came to cooperate, and our workload actually increased many times, so we were very busy. Zhao Minghui introduced that they used the platform established by cloning "Ping'an" to make some gene-edited cats, and also used the results to establish a connection with the enterprise and promote the transformation of some achievements in the laboratory.
If 'Ping'an is not born, the progress of the transformation of results may be slightly slower. But now many companies have taken the initiative to come to the door and cooperate with us very happily. Zhao Minghui introduced that he is now committed to the efficient reproduction of pets, the design and manufacture of disease models.
Zhao Minghui said, "When I was admitted to graduate school, I wanted to make cloned animals, and now I have achieved part of my dream." For Zhao Minghui's team of cloned cats, "Ping'an" will no longer play a research role, and it will live a carefree life in the future.
The team leader Zhao Minghui was born in 1986, and he led a group of "post-90s" to carry out scientific research.
The team has long been committed to canine and feline somatic cell cloning and disease model preparation. In the past 5 years, systematic research has been carried out on issues such as canine cloning, estrus determination, in vitro culture and maturation of eggs, and 16 SCI articles have been published in international mainstream academic journals as the first or corresponding author. Zhao Minghui has successfully cloned a number of elite working dogs, and in 2018, he took the lead in creating an in vitro culture method for early embryos in dogs, which has laid an important foundation for the research of canine reproductive physiology. The technology and experience accumulated by the team in the field of canine cloning provided an important technical guarantee for the smooth birth of the "Ping'an" cloned cat that year.
In addition to solid academic accumulation, the spirit of continuous pursuit of innovation also keeps this young team alive. Zhao Minghui said, "Behind the achievements is the high attention that the school attaches to us young talents. With the support of the school's active academic atmosphere and good scientific research conditions, the team continues to explore the construction of the best scientific experiment environment, "We have installed a variety of smart home devices in the laboratory, including temperature, humidity, lighting, smoke alarm, environmental control systems and face recognition systems, etc., so that the experimental environment can be intelligently controlled." Zhao Minghui said. "In the next step, we will focus our research on the gene editing of cats and carry out the cause of 'cat petting' to the end. ”
What exactly is the point of cloning animals?In this regard, Zhao Minghui told reporters that the general public's misunderstanding of cloning technology may be more from some foreign blockbusters, in fact, in the United States now, the beef on the table may be cloned, and the scientific research team will select the best cattle for cloning to produce more delicious beef.
In terms of cloning pigs, because the internal organs of pigs are similar to those of humans, after certain gene editing, the organs of the pigs, such as the heart, liver, and kidneys, can be used in the human body. This is called 'xenotransplantation', and it is also the focus of research now, because after all, there is still a need for a large number of transplanted organs in medicine. Zhao Minghui said, "Another aspect of the study of cloned pigs is reproduction, which can improve production efficiency to a certain extent." ”
In Zhao's view, animal cloning is not an ethical issue. Cloning technology is different from gene editing in that it does not remove the gene pool of the species. It is a misconception that cloning technology leads to gene damage and the insertion of foreign genes. In addition, cloning technology will play an important role in the pet industry and in facilitating the development of animal disease models and drugs. "And for endangered wild animals, cloning technology will also be helpful, you can collect its cells for cloning, then the next generation will be a new and fertile child, so that you can expand the population through mating." Zhao Minghui said.
Reporter:Technically speaking, what are the main difficulties of cloning technology?
Zhao Minghui:When it comes to cloning technology, most people may think that it is difficult to operate in the laboratory. In my opinion, the most important thing in the whole experiment is the breeding and management of animals, which many people may overlook, which is why many laboratories cannot develop cloned animals. At this point, I would like to thank my team, some of whom have Xi in animal hospitals and are more experienced in animal husbandry and management, which makes up for the shortcomings of research.
Reporter:Will the clone carry the memories of the original?
Zhao Minghui:Memories are acquired. The animals we clone do not contain proper memory. Pet owners will say that after getting the cloned cat themselves, they will raise it and feel the same as the original cat. In this case, it is not that the cloned cat has a memory, but because their genetic material is the same, resulting in the same personality, so the pet owner will have a sense of déjà vu for the cloned cat.
Cloning technology is widely used in the field of pets, and cloning cats and dogs have begun to enter the market in recent years. In the case of pets entering their twilight years or just dying, the cell state can be restored to the animal state through animal cloning technology, and the genetic material can be kept consistent with the original organism, so that the pet owner can find spiritual comfort.
Reporter:What progress has been made in cloning technology to protect endangered species?Will you and your team experiment with other species in the future?
Zhao Minghui:There is still a lot of room for development in cloning technology in the conservation of endangered species. Taking cats as an example, there is currently an operational idea that considering the close relationship between cats and tigers, cat eggs can be used to combine with somatic cells of Siberian tigers, and Siberian tigers may be cloned after transplantation. Our team currently does not plan to clone other species, after the establishment of the cloned cat platform, more want to use the platform to do some clone cat gene editing work, as an agricultural college, pet disease research is also the focus of our attention.
* Qingdao Morning Post reporter Zhong Shanglei **Courtesy of Qingdao Agricultural University.