How Dire Wolf that had become extinct 13,000 years ago have been brought back to life

History has been created as three pups of the Dire Wolf, a giant wolf species that had become extinct some 13,000 years ago from the face of the earth, has been brought back to life in a glorious feat of genetic engineering by an American company Colossal Biosciences. Salute!

Chances are you must have seen the famous American fantasy drama HBO TV series ‘Game of Thrones’, based on the novels of George Martin. After all, over one billion people out of the world’s eight billion people have seen it! Set on the fictional continents of Westeros and Essos, it follows a web of political conflicts among the noble families either vying to claim the throne or fighting for independence from whoever sits on it. In this fantasy series, you find an extinct animal called the Dire Wolf that has; now hold your breath, come back to life!

How Dire Wolves became pop-culture icons

Just recall that romantic setting in Game of Thrones: a frozen forest, a noble family called the Starks, and a litter of orphaned wolf pups stumbled upon them like a gift from the gods. In Game of Thrones, Dire Wolves aren’t just pets—they’re symbols, protectors, and extensions of the Stark children’s souls. They are the sigil (an inscribed or painted symbol considered to have magical power) of the Starks. They are no ordinary wolves (grey wolves); they are Dire Wolves.

Now, Martin had not just whip up Dire Wolves from a cauldron of pure imagination. Dire Wolves (Aenocyon dirus) were actually real beasts—massive, muscular wolves that roamed the Americas from about 125,000 to 10,000 years ago. Much bigger and brawnier than today’s grey wolves, they were apex predators with very strong jaws built to crush bone. Martin gave these real-life majestic animals an emotional twist—powerful, protective, fiercely intelligent, and bonded to humans.

Dire Wolves took deep roots in public imagination because in the Game of Thrones, each pup bonds with a Stark kid, mirroring their personalities and fates. There’s Grey Wind, fast and ferocious like Robb, the young king; Lady, gentle and doomed like Sansa; Nymeria, wild and untamed like Arya; Summer, brave and watchful like Bran; Shaggydog, chaotic like little Rickon; and Ghost, silent and mysterious like Jon. These Dire Wolves fight; rip out throats; guard their humans from assassins; and even lead wolf packs in the wild after being separated. They’re not invincible, though—some meet tragic ends, reflecting the brutal stakes of the story. Imagine them as furry guardian angels with teeth, stalking through a tale of ice and fire. No wonder, people liked them so much.

The miracle that happened

Just a few days back, Colossal Biosciences Inc., an American biotechnology and genetic engineering company, revealed that six months ago, they had ‘created’ three genetically engineered Dire Wolf pups, christened Romulus, Remus, and Khaleesi. Naming them Romulus and Remus is a nod to Roman mythology. The twin founders of Rome by these names are believed to have been suckled by a she-wolf after being abandoned. You may recall that in Rudyard Kipling’s memorable novel ‘The Jungle Book’ also, the boy Mowgli is raised by a pack of wolves in an Indian jungle after being orphaned as a baby. Millions of people can still hum the title song ‘Jungle jungle baat chali hai pata chala hai; chaddi pahan ke phool khila hai’ of the hugely popular TV series based on it.

Naming the genetically engineered pups Romus and Remulus is a juicy hint—perhaps the scientists wanted to suggest that the legendary she-wolf of Roman mythology was a Dire Wolf, a prehistoric titan nurturing the story of Rome’s origin. It’s not a stretch. Real Dire Wolves roamed Europe long ago too, and their size fits the mythic vibe. The third pup Khaleesi, a female, gets her name from the dragon queen in Game of Thrones.

If you recall, the 1993 blockbuster film Jurassic Park had a similar theme. A wealthy businessman and a team of genetic scientists ‘created’ dinosaurs and kept them in a wildlife park on an island near Costa Rica. In fact, most people learnt the word de-extinction for the first time in that film. De-extinction (also known as resurrection biology, or species revivalism) is the process of generating an organism that either resembles or is an extinct organism. Following Jurassic Park, there have been five more films in the Jurassic Park franchise, the last one coming in 2022.

“They’re beautiful, they’re cute, they’re incredible,” said Ben Lamm, co-founder and CEO of Colossal Biosciences, sounding every bit the enthusiastic new dog parent. “We took a 13,000-year-old tooth and a 72,000-year-old skull, and made puppies.” Wonderful!

Motivation behind the project

Colossal Biosciences, a Dallas-based biotech company founded in 2021 by Ben Lamm and geneticist George Church, aims to “make extinction optional” through advanced genetic engineering. Their mission blends conservation with profit-driven innovation, targeting species like the woolly mammoth, dodo, and the Dire Wolf (Aenocyon dirus), which went extinct some 13,000 years ago. The Dire Wolf project, unveiled on April 8, 2025, was motivated by a desire to showcase their “end-to-end de-extinction technology stack” and prove its potential for both resurrecting lost species and aiding endangered ones, like the red wolf. The Dire Wolf, a culturally iconic predator from North America (popularized by Game of Thrones), offered a compelling proof-of-concept due to its close genetic relation to the grey wolf (Canis lupus) and abundant fossil record.

The fossils of the Dire Wolf

In 2021, a separate team of scientists managed to retrieve DNA from the fossils of Dire Wolves.  Colossal sourced DNA from two Dire Wolf fossils: a 13,000-year-old tooth, excavated from Sheridan Pit, Ohio, part of a natural history museum collection; and a 72,000-year-old ear bone, found in American Falls, Idaho, identified as a petrous bone (part of the skull near the ear). These samples were chosen for their dense structures—teeth and petrous bones are known to trap and protect DNA better than other bones due to their low porosity and hydroxyapatite matrix. Marvel at the antiquity of the fossils and you will have nothing but respect for the work. 

How the DNA was extracted from the Fossils?

Extracting ancient DNA (aDNA) from these fossils involved meticulous lab work:

• Cleaning: Fossils were UV-irradiated and sandblasted in a clean room to remove surface contaminants.
• Sampling: Powder was drilled from the tooth’s dentin and the petrous bone’s cochlear region—areas rich in cellular DNA.
• Decalcification: Samples were soaked in EDTA to dissolve mineral content, releasing DNA.
• Purification: Using a silica-based method, DNA was isolated from inhibitors like tar or soil residues.
• Sequencing: Next-generation sequencing (NGS) produced short reads (20-100 base pairs), assembled into genomes with 3.4-fold coverage (tooth) and 12.8-fold coverage (ear bone), far exceeding prior Dire Wolf data.

The petrous bone yielded higher-quality DNA due to its density, while the tooth’s DNA, though fragmented, was sufficient for mtDNA and nuclear sequences. Damage patterns (e.g., cytosine deamination) confirmed authenticity.

The gene editing technology used for ‘creating’ the pups

In simple words, the Colossal Biosciences researchers edited 20 genes of grey wolves to imbue the animals with key features of Dire Wolves. They made 20 strategic edits across 14 genes, with 15 of the edits recreating extinct gene variants. The nuclei were then removed from the resultant cells and transferred into donor egg cells, creating embryos. These embryos from the edited grey-wolf cells were implanted in surrogate dog mothers and waited for them to give birth. Forty-five embryos were cultured in vitro, then implanted into two surrogates. Surrogates were chosen for size and health to handle large pups. One embryo per surrogate succeeded. Just as in human fertilisation treatments, not every embryo resulted in a pregnancy. Colossal made multiple attempts in an effort to increase the odds of success, completing eight embryo transfers, involving 30 to 45 specimens each.

Since Dire Wolves are extinct, Colossal used domestic dogs (large mixed-breed hounds) as surrogates. This wasn’t cloning with pure Dire Wolf DNA but a hybrid approach, rewriting grey wolf genes to mimic Dire Wolf traits. Brilliant!

They used CRISPR-Cas9, a precise gene-editing tool, to modify grey wolf DNA. CRISPR stands for Clustered Regularly Interspaced Short Palindromic Repeats.  The system consists of two key components: the Cas9 enzyme and a guide RNA (gRNA).

And you got a Wolf that disappeared 13000 years ago

The result was three healthy and extremely cute wolf pups—two males that are 6 months old and one female that is 2 months old. Romulus and Remus (males) were born via C-section on October 1, 2024, after 65 days gestation. Khaleesi (female) followed on January 30, 2025, from a third surrogate. The pups were bottle-fed after a few days to manage surrogate bonding. They are big, for one thing, and have dense, pale coats not found in grey wolves. Colossal is keeping the wolves on a private 2,000-acre facility at an undisclosed location in the northern United States.

Some scientists argue they’re “hybrid animals and not true Dire Wolves. Others call them “functional copies” of Dire Wolves, emphasizing phenotypic similarity over genetic purity. In any case, Dire Wolves and grey wolves share 99.5% of DNA. It is only the 0.5% that makes all the difference.

What this research means to the world

Colossal’s team extracted and sequenced Dire Wolf DNA, assembled genomes, then compared the results to that of living canines. This enabled them to pinpoint where Dire Wolves’ genomes differ from that of other species. They also discovered that grey wolves share 99.5% of their DNA with Dire Wolves, making them the extinct animal’s closest living relative. The methodology used was similar to the one in their woolly mouse project. Revealed in March, Colossal’s woolly mice were created by editing the genome of normal mice to display physical traits similar to those found in woolly mammoths—particularly their long, fluffy fur. Colossal states that these three wolf pups are proof that a “standardized toolkit for de-extinction” is possible.

Inferiority complex in Indian media

The most pathetic state of research in India is well-known. There is no significant invention or discovery in the world that could be traced to Indian talent working in India since independence. This has led to a huge inferiority complex amongst Indians. Things have reached such a deplorable pass that instead of appreciating this tremendous scientific achievement, a section of the Indian media has ridiculed it. In an editorial titled ‘The DeExtinction Con’, a paper wrote that it’s about human entertainment, not animal conservation! It also lamented the fact that the pups are said to be enjoying a Ritz Carlton lifestyle (that is, replete with luxury experiences, personalized service, and exquisite environments). This is not just ridiculous; it is a classic Freudian slip. Little realizing that the pups born out of ancient DNA do need extra care to monitor their growth, behaviour, and the impact of the genetic edits, ensure their welfare, and plan for their future, including potential re-wilding efforts, these intellectually bankrupt people are making fun of it.

By God, this is sci-fi fairy-tale of de-extinction having become a reality. A 72,000 year old bone fragment yielding a full living body! And, instead of marvelling at it or citing it as an example for Indian researchers to take inspiration from, Indian media ridiculed it. Pathetic! No wonder, Indian science is doomed.