Colossal Biosciences has successfully resurrected the dire wolf (Aenocyon dirus), an apex predator that has been extinct for over 10,000 years.

This pioneering project signifies the first successful de-extinction of a species and redefines the possibilities for wildlife conservation.

The team of scientists utilized cutting-edge DNA analysis, CRISPR gene editing, and innovative cloning techniques to carry out this extraordinary feat.

They extracted genetic material from two ancient dire wolf fossils: a 13,000-year-old tooth discovered in Ohio and a 72,000-year-old skull found in Idaho. Unlike findings from the La Brea Tar Pits, where asphalt degradation compromised DNA, these ancient remains provided sufficient genetic material for sequencing.

Through advanced methods, the researchers generated 70 times more genomic data than previous studies, achieving a 12.8-fold coverage of the dire wolf genome. Their analysis revealed a striking 99.5% genetic similarity between dire wolves and modern gray wolves, indicating they diverged evolutionarily over 5.7 million years ago.

By comparing the genomes of dire wolves and gray wolves, the scientists identified 20 critical genetic variants across 14 genes responsible for traits unique to dire wolves, such as their larger size and distinct white coat.

To mitigate potential health risks linked to pigmentation genes, the team selectively edited genes like MC1R and MFSD12 to safely recreate the iconic light coat.

In an innovative cloning breakthrough, the team developed a new method using endothelial progenitor cells (EPCs) sourced from gray wolf blood, a less invasive alternative to traditional tissue sampling.

These cells were edited with dire wolf variants and implanted into denucleated gray wolf eggs, which were then transferred to surrogate domestic dogs, resulting in three healthy pups named Romulus, Remus, and Khaleesi.

The successful revival of the dire wolf represents a significant step towards addressing biodiversity loss, with Colossal’s techniques integrating ancient DNA sequencing, multiplex gene editing, and EPC-based cloning—paving the way for the revival of other extinct species and the enhancement of endangered populations.

According to CEO Ben Lamm, the return of the dire wolf is “just the beginning” of utilizing synthetic biology for ecological restoration, marking a hopeful future for conservation efforts.