Designer Rams and Dire Wolves

At Colossal Biosciences in the Dallas area, three massive canids roam a private enclosure.Romulus, Remus, and Khaleesi appear at first like robust gray wolves. However, their pale coats, heavy jaws, and behavior suggest a deeper lineage. These are the world's first "functionally de-extinct" dire wolves.

"This is what we call functional de-extinction," said Matt James, Chief Animal Officer at Colossal. "It relies on cloning technology, but we are using genetic engineering to try to identify close living relatives to an extinct species.

The closest relative turned out to be the modern gray wolf. From there, scientists edited 20 key genes to recreate traits unique to Canis dirus, such as heavier bone structure, broader skulls, and metabolic characteristics aligned with the ancient apex predators.

"This isn't a Frankenstein experiment. We want to be very measured and deliberate and ensure animal welfare," James said. "Twenty edits was this nice area where we could get the core phenotypes in, but also ensure positive outcomes and optimal welfare for the animals."

James and his team wrote a 160-page animal care manual for a species that had been extinct for over 12,000 years. Their work involved sequencing a 13,000-year-old tooth from a museum in Ohio and a 72,000-year-old skull from Idaho, which provided 500 times more data than had ever been available on dire wolves.

"You can't just clone from 10,000- or 70,000-year-old tissues because the DNA is no longer viable," he explained. "There's a lot of genomic sequencing and computational biology required to piece all the broken fragments of DNA back together."

Although media coverage has generally referred to this as "cloning," there is much more going on here. There are two issues at hand: cloning and gene editing. Cloning involves creating a genetically identical copy of an organism. This process replicates the entire DNA sequence of the original, resulting in an individual with the same genetic makeup as the original. Gene editing, on the other hand, is a precise method of altering specific genes within an organism's DNA. Using tools like CRISPR-Cas9, scientists can add, remove, or modify genetic material to correct mutations, enhance particular traits, or study the functions of genes. Unlike cloning, gene editing doesn't create a copy of an existing organism but instead modifies its genome in targeted ways.

While dire wolves represent a case of deliberate, high-profile de-extinction, another case shows how the same tools can be used with far different motives.

Five-month-old dire wolf pups roam the 2,000-acre preserve in Texas. Courtesy Colossal Biosciences.

In 2024, Arthur "Jack" Schubarth entered into a plea agreement with federal officials for one count of Lacey Act trafficking and one count of conspiracy to violate the Lacey Act. According to Forest Legality, the Lacey Act, initially created in 1900, is a United States law that bans the trafficking of fish, wildlife, or plants that are illegally taken, possessed, transported, or sold. Schubarth and five others, who federal officials labeled in court documents as Persons A-E, were allegedly involved in various activities. Three are from Texas, one from Montana, and another from Minnesota.

Court papers cite the following claims:

• On or about January 25, 2013, Person A entered the United States with biological tissue from a Marco Polo argali sheep that had been hunted in Kyrgyzstan. Person A did not declare the animal parts upon entry.
• On or about October 6, 2015, Schubarth entered into an "Ovine Cloning Contract" to clone an unspecified number of Marco Polo sheep from the tissue. He provided a deposit of $4,200 for the cloning.
•  On or about November 22, 2016, Schubarth received 165 cloned Marco Polo embryos at the Schubarth Ranch.
• On or about May 15, 2017, a pure Marco Polo argali sheep male was born from the cloned embryos Schubarth named "Montana Mountain King."
• Starting in 2018 at the latest, Schubarth harvested semen from M1\.1K to inseminate ewes via LAP-AI to create Marco Polo argali hybrid offspring.

Other notes from the case involved sending semen straws from MMK to Texas as well as transferring its offspring to Texas. This has resulted in numerous Marco Polo sheep hybrids being bred, sold, and possibly hunted.

Montana Mountain King at the time of this writing is still alive. He was cloned from tissue illegally smuggled in from Asia. Courtesy Montana Fish, Wildlife & Parks Photo

Hybridization of wild sheep species is not a new concept. More than 50 years ago, Louis Stumberg of the Patio Ranch in Texas bred Mouflon ewes with Argali rams which at the time were present in a zoo to create what became known as the Stumberg Sheep.

The Texas Dall rams, which are popular in exotic hunting circles, for example, are a hybrid of a mouflon and merino, albeit a distant one. The difference now is these animals don't have to be put in a pen together and bred anymore. They can be cloned from simple tissue or be artificially inseminated with sperm from cloned animals. And the search is on for more tissue, with numerous social media posts showing a desire to get tissue from species like the Altai argali.

The Stumberg is a hybrid sheep breed created by a Texas rancher decades ago but it was done the old fashioned way-by putting animals in a pen and allowing nature to take its course. Photo by Chester Moore.

While cloning wildlife might be a shock to many within the hunting community, it has been a reality at the highest levels in the world of endangered wildlife management for several years.

On December 10, 2020, "Elizabeth Ann," a black-footed ferret, was cloned from the frozen cells of "Willa," a specimen that lived in the 1980s as part of a U.S. Fish & Wildlife Service project that involved scientists at Revive & Restore (a cloning conservation group), ViaGen Pets & Equine, San Diego Zoo Global, and the Association of Zoos and Aquariums.

"The Service sought the expertise of valued recovery partners to help us explore how we might overcome genetic limitations hampering the recovery of the black-footed ferret, and we're proud to make this announcement," said Noreen Walsh, Director of the Service's Mountain-Prairie Region. "Although this research is preliminary, it is the first cloning of a native endangered species in North America, and it provides a promising tool for continued efforts to conserve the black-footed ferret."

In 2024, the Service and its genetic research partners announced the birth of two more black-footed ferret clones – known as "Noreen" and "Antonia" both cloned from the same frozen specimen as Elizabeth Ann, who is alive and thriving at the ferret program's headquarters in Fort Collins, CO. Service officials said the application of this technology to endangered species addresses specific genetic diversity and disease concerns associated with black-footed ferrets.

"The Service views this new potential tool as one of many strategies to aid species recovery alongside efforts to address habitat challenges and other barriers to recovery."

In February 2023, a cloned Przewalski's horse, a native of Mongolia, was born at the ViaGen Pets & Equine cloning facility in Texas and was moved to the San Diego Zoo Safari Park. According to San Diego Zoo officials, "Ollie" is a clone of a male Przewalski's horse stallion whose living cell line was cryopreserved over 40 years ago in the San Diego Zoo Wildlife Alliance Biodiversity Bank's Frozen Zoo.

"The world's first cloned Przewalski's horse, Kurt, also living at the Safari Park, is the genetic twin of the foal, as he was born in August 2020 from the same stallion's living cell line."

Although ferret cloning is groundbreaking in several ways, it is not the first cloning of a wild animal at a governmental level. It happened with wild sheep back in 2015. According to The Guardian, researchers at Iran's Royan Institute cloned a wild Isfahan mouflon using a domestic sheep as a surrogate mother.

"We have been working on the project for around four years," project head Mohammad Hossein Nasr-Esfahan, told the Guardian. "Conservation of wildlife is an important concept in developing countries, and so far, a few successful projects involving the birth of wild animals by interspecies cloning have been achieved worldwide."