New advancements in cloning.

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‘DE-EXTINCTION’ CLONING

Scientists have been on a mission to ‘resurrect’ extinct species of animals, as part of ‘de-extinction’ initiatives that seek to ‘create new versions of lost species’.

The first extinct species was cloned in 2009. The bucardo, a type of Spanish mountain goat, went extinct in 2000. The bucardo was cloned using goats as egg donors and surrogates yet died soon after birth.

Now, ‘breakthroughs’ are being met where the cloned animals are surviving past their birth.

Maya, the first successful arctic wolf clone, turned 100 days old recently, after being bred by bioengineers at Beijing’s Sinogene Biotechnology.

The cloned wolf made her debut in September 2022 at Harbin in the Heilongjiang Province of China, with her beagle surrogate mother.

“After two years of painstaking efforts, the Arctic wolf was cloned successfully. It is the first case of its kind in the world.”

Maya’s donor cell came from the skin sample of a wild female arctic wolf also named Maya, who died at 16 years old, which is considered old age for a wolf.

Sinogene took genetic material from Maya 1.0 and created 137 embryos using eggs from female dogs.

They then implanted 85 of those eggs into 7 beagle surrogates.

In July of 2022, ‘Maya 2.0’ was born.

Currently, she lives with her beagle mother at Sinogene. She may have to live there for the rest of her life, because she didn’t have early socialisation with other wolves and therefore couldn’t live in the wild.

“The newly born wolf has the same genome as the original wolf, but the cloned wolf hasn’t lived with other wolves, but with a dog,” said Zhao Jianping, Sinogene’s Deputy General Manager.

It isn’t just artic wolves that bioengineers are fascinated with ‘resurrecting’.

In fact, one of Australia’s most iconic extinct species is also on this list: The Tasmanian Tiger.

In the late-19th century, European colonisers in Tasmania blamed the dog-sized, tiger-striped, carnivorous marsupial for killing their sheep and chickens, and slaughtered thylacines by the thousands.

The last known thylacine spent its days pacing a zoo cage in Hobart, Tasmania, and died of neglect in 1936. Now the Tasmanian tiger is poised to become an emblem of de-extinction.

Australian scientists have been hoping since 1999 to use emerging genetic technologies to try to bring the thylacine back from the dead.

In March, a Melbourne University team established the Thylacine Integrated Genetic Restoration Research Lab with a philanthropic gift of five million Australian dollars (about $3.6 million).

They are using DNA from a preserved thylacine pup (pictured right) that has been stored in hopes of new advancements in technology one day.

Now, with new technologies at their disposal, along with the de-regulating of gene-editing techniques in Australia, the scientists are ‘working hard’ to ‘bring back’ such a national icon.

And this is not all.

Bioengineers are looking even further back in their efforts to ‘de-extinct’ species with technology. Back to ‘pre-historic’ ages where massive creatures walked the plan. Creatures like Woolly Mammoths.

One Texas de-extinction company, Colossal Biosciences, is working toward just this goal.

The CIA is investing in the biotechnology company in an effort to see Woolly Mammoths be resurrected from the dead and bring ‘thunder upon the tundra’ once again.

In-Q-Tel, Colossal’s new investor, is a non-profit venture capital firm that receives funds from the CIA.

In a blog post in late September, the company shared why it decided to become a Colossal investor.

“Why the interest in a company like Colossal, which was founded with a mission to ‘de-extinct’ the woolly mammoth and other species? Strategically, it’s less about the mammoths and more about the capability.”

Along with the CIA, others who have already invested in the Dallas-based company are Peter Thiel, Tony Robbins, Winklevoss Capital and more. 

Colossal Founder, Ben Lamb, spoke about the company’s plans to bring back the woolly mammoths in an email to media outlets:

“Biotechnology and the broader bioeconomy are critical for humanity to further develop. It is important for all facets of our government to develop them and have an understanding of what is possible.”

The concept of ‘de-extinction’ sciences is a new evolution in cloning technologies that have been around for many decades, and if mastered, could eventually move to humans as well — if it hasn’t already.

CLONING ‘REVOLUTION’

The cloning process, in its simplest form, involves producing individuals or cells with identical DNA, either naturally or artificially.

Cloning, in fact, is a natural form of reproduction used by plants, fungi and bacteria that has enabled life forms to spread for as long as known time.

In contrast, scientists have been able to tap into this process to artificially create clones.

According to the NHGRI, there are three different types of artificial cloning:

• Gene cloning: This type involves creating copies of genes or DNA segments.
• Reproductive cloning: This type involves making duplicates of whole animals.
• Therapeutic cloning: This type involves creating embryonic stem cells.

The roots of cloning began in the nineteenth century.

In 1885, German biologist Hans Driesch showed that blastomeres of two-cell sea urchin embryos could be physically separated and two entire embryos formed from each blastomere.

This was the first-ever demonstration of ‘artificial embryo twinning’, the process of separating embryonic cells in early stages of development. Each separated cell continues to grow/be implanted into a surrogate.

Then, in 1902, German embryologist Hans Spemann used this same method to clone salamanders.

Interestingly, the two cells of a salamander embryo are much stickier than sea urchin cells. Therefore, Spemann created a tiny noose from a strand of baby hair and tightened it between the cells until they separated. Yes, I am not making this up.

Sticking with his noose and salamander, Spemann in 1928 essentially performed the first instance of nuclear transfer. At the 16-cell stage, Spemann used his noose to separate a cell from the rest of the embryo. This single cell grew into a new salamander embryo.

This experiment demonstrated that the nucleus from an early embryonic cell could direct the complete growth of a salamander.

In 1953, Robert Briggs and Thomas King (Figure 1) were able to perform the first successful nuclear transfer by transferring the nucleus from an early tadpole embryo into an enucleated frog egg.

This led to the development of a tadpole.

In 1958, British biologist John Gurdon was the first to successfully clone an animal using somatic cell nuclear transfer. In this approach, the nucleus from a somatic cell is removed and placed into the enucleated egg cell of another animal.

Gurdon transplanted the nucleus of a tadpole intestinal cell into an enucleated frog egg. He was able to create tadpoles that were genetically identical to one from which the intestinal cell was taken.

This was one of the first major steps towards cloning.

Established in 1993, The Roslin Institute is an animal sciences research institute, part of the University of Edinburgh. In 1996, at the Institute, British scientists Ian Wilmut and Keith Campbell transferred the nuclei from cultured cells into enucleated sheep egg cells.

All previous cloning experiments used donor nuclei from cells in early embryos.

The lambs born from this experiment were named Megan and Morag and paved the way for Dolly.

This experiment showed that cultured cells could supply donor nuclei and that it might be possible to use modified cells to create transgenic animals.

Then in 1996, in a landmark experiment, Wilmut and Campbell created the first mammal by somatic cell nuclear transfer. A nucleus from an adult sheep’s udder cells was transferred into an enucleated egg.

Of the 277 attempts, only one produced an embryo that was carried to term in a surrogate mother.

The famous lamb, named Dolly (after Dolly Parton), brought cloning to the forefront and into the public light (Figure 2). This experiment showed that differentiated cells, expressing only a distinct subset of genes, could be redesigned to grow an entirely new organism.

This changed our understanding of a basic scientific principle.

Multiple other animals have been cloned since the experiments at Roslin, from mice to camels.

Cloning in popular culture is often depicted as something negative. From bringing back dinosaurs using DNA extracted from fossils, to cloning human counterparts for organ donations, science fiction films have used the concept of cloning to emphasise the dangers associated with such scientific developments.

With everything in popular culture, these portrayals reflect societal fears and questions surrounding advanced technology.

The news of Dolly the sheep and its widespread media attention, captivated the world and made many people question – what if we could clone humans?

Apart from conducting these experiments to show that it was possible and to refine techniques, cloning also has several commercial applications.

The initial hopes were to use cloning to produce animals that made human protein in the milk and blood of transgenic cloned animals.

To date, several transgenic and cloned animals have been harvested in an experimental setting to produce different human proteins, including the human coagulation factor IX, human anti-thrombin and Alfa-1-antitrypsin. This approach, however, has been largely superseded by other more large-scale methods.

One of the most well-known and controversial forms of human-animal cloning involves chimeras.

Recent advances in stem cells and gene engineering have paved the way for the generation of interspecies chimeras, such as animals bearing an organ from another species.

Human-animal chimeras provide the ability to produce human organs in other species using autologous stem cells which would be patient-specific and immune-matched for transplantation.

Researchers have expressed ethical concerns over the practice, including “…the risks of consciousness and of human features in the chimeric animal due to a too high contribution of human cells to the brain, in the first case, or for instance to limbs, in the second.”

Conversations about human cloning often relate to fears of eugenics and loss of human individuality.

Allopathic medicine and the advancements of science are pathed with a bizarre history.

Are these the types of people we want in charge of steering humanity’s future?

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