Bringing Back the Dead: Mammoths pt. 3: A colossal challenge…

In the not-too-distant future.

Echoes abound down the long hallway. Several scientists emerge from the changing room, each wearing identical lab coats, along with gowns, goggles, masks, and gloves. Their chatter dies down as they open the doors into the main chamber, and they are hit with a wave of warm air. The air inside the isolated ward is kept at a stable 35.9o.

Already some of the scientists begin to sweat. If this is because of the temperature, or because of what lay inside the room, it’s too hard to tell. As they step into the room, the lead scientist looks up at the most prominent of the cameras hanging from the ceiling, and she begins to speak, knowing that every word will be recorded for posterity.

“We are here today to mark the birth of Experiment zero-one-one-B, and the return of an organism once thought lost from our planet.” She leads the rest of the scientist through rows upon rows of vacuum sealed plastic containers nearly as tall as them. Each is hooked up to a series of tubes, valves, diagnostic devices, and monitors, along with a heavy skeleton of supports.

If any of the scientists paused to look into the bags and peer through the milky fluid inside, they would see embryos in various states of gestation. The deeper they walk into the room, the more the embryos take shape, starting out as vague curled up rectangles, but becoming something more recognizable, more distinct, more developed, and hairier. Some of the forms and figures are moving, wiggling and shaking the bags from the inside.

“Today we welcome back into the world, a creature who has not roamed the Earth for more than four thousand years.” The head scientists has paused outside one of the largest, most bulging, plastic bags. Its contents obscured by the plastic coating, the figure inside is struggling against the container, as though it is fighting to get out. A camera has been set up in front of this container, and another scientist, sweating and poised, focuses first on the bag and then on the head scientist as she speaks. “With a few more throws of life, lets welcome zero-one-one-B, or Betty as we are calling her, into the world.”

The camera tactfully focuses on the head scientist while the others go about separating various hoses and connections. The creature inside struggles harder, powerful limbs kicking forth with a force and a weight almost surpassing any of the scientists. The camera zooms in on the fresh face, as its eyes open for the first time, closing quickly, as the creature thrashes about. There had been a discussion about using a sedative during the birth, but the risk was deemed too great after what had happened with zero-one-zero-A or Alonzo. So the scientists did their best to remove all of the equipment without getting kicked.

Finally, the last gasket was separated and the skeleton, on hydraulics, was lowered so that the animal inside the bag was near the artificial ground set up to catch the infant as it landed. With a quick nod from the head scientist, another scientist reached out and grabbed a chord on the bottom of the bag, duking a heavy foot swinging inside. The chord had been attached last night, when they had determined that it was time for the baby’s birth, and as the scientist stood, he yanked on it, pulling from corner to corner separating the bottom of the bag.

The milky fluid spilled out of the bag, washing into the grates set into the floor to drain it away, flowing out with the liquid was the massive harry form of Betty, who landed, still thrashing, atop the gentle pallet meant to simulate the hard ground upon which she would have naturally been born. “Thud.”

Stunned by either the short fall, her first entrance into the real world, the heavy creature stopped thrashing for a second. Another second, and Betty did not move at all. Anxiously the scientists waited, the head scientists contemplated telling the cameraman to cut the feed. But before she could utter a word, the Baby coughed up a small bit of liquid and heaved.

For the first time 4,000 years, a Woolly Mammoth took its first breath of fresh air.

End

Not my best work of art, but every now and then I want to remind all of you how lucky we all are that Bailey does all of the illustrations for my book instead of me. His name is Barnabas the Mammoth, and I like him. Wouldn’t it be nice to share the steppe with him one day?

In 1993 a movie came out exposing the world to wonderful new possibilities. Animals long dead may rise again!

The movie bringing this idea to the forefront of popular thought was, of course, Universal Picture’s We’re Back A Dinosaur Story. We're Back! A Dinosaur's Story (film) - Wikipedia. In which a T-Rex, a Parasauralophus, a Triceratops, and a Pterosaur (which is not a dinosaur) are all brought into the future by time traveling aliens (and humans) hoping to deliver them to the Museum of Natural History in 1993 New York. Hijinks ensue, and after the dinosaurs (and pterosaur) get an amped up intelligence from a special cereal, they end up becoming functioning members of 1993 Earth Society, playing golf, rampaging through the Macy’s Day Parade while impersonating animatronics, and saving human children from the inverse cereal which lowers their intelligence.

This movie opened up a brand-new potential to the world. All we would have to do is team up with aliens, go back in time, invent a brain growing cereal, and ask dinosaurs what life was like in the Mesozoic!

I’m kidding. There was another much more influential movie which came out about dinosaurs and also opened the world to new possibilities in 1993. Carnosaur. Actually, I jest again. This horror movie is #3 on the list of my Top Dinosaur Movies to never recommend to anyone, ever. An entirely made-up list, that was developed just to put Carnosaur into the corner it so rightly earned.

No. The movie I’m actually referring to is Jurassic Park based on the 1990 novel written by Michael Crichton (No link provided here because, duh.) In 1993 genetics and biotechnology were broaching new frontiers. Cloning was actively being discussed. The potential in understanding and sequencing DNA was being contemplated, and technology was becoming advanced enough to help us peer deeper into that genetic mess of information and also to allow us to record and store those bits and pieces of information digitally.

This was all as new and exciting as discovering that the computer you needed to operate to save your life was a Unix system. “I know this.” – Lex Murphy, Jurassic Park. And while digital technology was becoming advanced enough to help us delve into the mysteries of DNA it was also helping moviemakers in Hollywood leap to the forefront of what it was possible to put on screen.

While movies like Carnosaur were reliant wholly upon animatronics, and We’re Back A Dinosaur Story was entirely animated, Jurassic Park had some of the best animatronics ever, but it also had some of the best digital effects possible! We could finally, digitally, able to put a Brachiosaurus atop a Hawaiian hill! But what if we want to put a real brachosaurus on that Hawaiian hill?

The same computers which allowed movie makers to put a Giant Sauropod on screen, also promised the potential to look at, and potentially eventually, sequence that Sauropod’s DNA… if only we had a source. (not the exact same computers obviously).

This burgeoning of technology and genetic understanding referenced in Jurassic Park wasn’t just about our growing understanding of DNA. The movie was predicated upon another merging of DNA and technology. An idea long thought science fiction but discussed, finally, as potential science fact. Cloning.

In 1993 we were still a few years away from the first mammal cloned from adult cells: Dolly the sheep. But the methodologies to harvest DNA and use it to copy critters were already being discussed. (and technically cloning had already happened, but it wasn’t headline grabbing because it was stem cell cloning.)

Jurassic Park simply took discussions of DNA , technology, and cloning to their logical conclusion: Let’s bring back dinosaurs.

As it turns out, the movie was a little off. The dinosaurs, accurate at the time, now are a little lacking comparative to our modern understanding of them. Also, we have never found a good source of Dino-DNA.

Bringing back dinosaurs is probably a pipe dream. Studies of extinct moa bones from New Zealand fixed the potential preservation of DNA under optimal conditions at 6.8 million years. https://www.popsci.com/science/article/2013-02/whats-half-life-dna  calculated from that 521-year half-life discussed in the previous article. 6.8 million is a bit shy of the + or – 65 million years necessary to store any Cretaceous Dino-DNA. (much less their Jurassic Ancestors like the stegosaurus which are further separated by time from the T-Rex than we are)

But whereas Jurassic Park fed us the inaccurate hope of using Dino-DNA stored inside the stomach of Jurassic, (and mostly Cretaceous) era mosquitos trapped in Jurassic, (and mostly Cretaceous) era amber being used to resurrect the coolest creatures ever to walk the Earth, the potential to bring back much more recently exticntified animals was also being talked about.

Animals like the Thylacine, (The Tasmanian Tiger,) which had only gone extinct less than 60 years before (from 1993.) Perhaps it could be brought back. People had preserved several bits of the last remaining marsupial tiger-like creatures for posterity. Or another of another recent, (but not as recently,) extinctified animal: the Dodo.

The resurrection of the Dodo was mentioned alongside dinosaurs in one of the most pivotal scenes of sea-bass adjacent discussion ever portrayed on film. In Jurassic Park, John Hammond and his small team of Dino-experts sat around the table to discuss the merits of of not only bringing back dinosaurs, but also the possibility to bring back organisms which humans directly had a hand in bringing to, or past the point of, extinction.

If a method could be found to clone animals, then perhaps it could bring back the Dodo. Perhaps it could bring back the Thylacine. Perhaps it could even bring back the Dino. In 1993 all of these seemed possible. Dinos, as previously mentioned are a little more challenging to bring back then posited by Crichton. And I will leave more of that discussion to another day, lets focus instead on the title of this series of blogs: Mammoths.

Mammals had been cloned before Dolly. The first cloned sheep were generated from embryonic stem cells in 1984. But in 1996, a new method was used to bring Dolly into the world. The nucleus from an adult sheep cell was inserted into an unfertilized egg, and the new embryo was carried to term in another sheep mother. This method meant that we could use DNA from developed ‘adult’ cells to produce clones.

For organisms like the Thylacine and the Dodo, this meant there were still a few more challenges. We could, potentially, harvest Adult DNA from cells in preserved remains of either animal, but the Dodo is a bird, so any fertilized embryo would have to be carried to egg in a close relative of a Dodo. Considering Dodo’s closest living relative is a pidgeon or dove, there’s a bit of a size issue. Dodo eggs were about 9 inches in diameter, which is over twice the size of a pidgeon egg. (yeouch)

We might be a little nearer with the thylacine, it’s a mammal, (a marsupial, but still,) so there’s no egg problem, but it’s closest living relative is the Numbat. (An animal I did not know existed until starting this article.) But there’s estimated to be about 5% difference between Thylacine DNA and a Numbat’s DNA. For reference, Humans and chimps share about 99% of our DNA. So, there’s still a bit of a hurdle there. Also the numbat is quite a bit smaller than the thylacine was, has an entirely differn’t behavioral pattern, and there are other problems.

So what about two animals which share even more DNA? Some kind of mammal which, while extinct, shares enough DNA with something still alive that if both were still around, then they might even be capable of producing viable hybrid offspring?

Mammoths meet all the hypothetical requirements for De-extinction. They are Extinct. (Duh) They may have been brought to extinction by man. (See part one in this series.) Their DNA has been preserved in “ideal conditions.” (See part 2 in this series.) And they have a living relative. 2 in fact. The African Elephant and the more genetically similar Asian Elephant. They also broach a 4th made up category, as wooly mammoth toys are often boxed in with dinosaurs, so despite the difference in age they also meet the prehistoric cool factor.

So, we have an extinct animal, whose DNA has been sequenced, and which shares, (according to some) 99% of the DNA with a modern relative. It may, or may not, have been killed off by man. But it is cool.

Mammoths. Can we bring them back?

Some people want to.

As early as 2008 articles began to appear discussing plans to bring back the wooly mammoth. It was a colossal idea. So colossal in fact that Colossal became the name of a company founded with just such an aim in mind.

(Sci-fi had already brought up the idea of de-extincting Mammoths using various cloning methods before 2008. But it is Colossal upon which I turn our focus, primarily because they have made themselves the primary voice behind the process of bringing the Mammoth back from extinction.)

In 2008, The idea of bringing back the mammoth didn’t seem that far away. In a time still brimming with ideas about the potential for DNA, cloning, and other such fun, it even seemed easily within reach, Maybe only a couple years away.

We had the right ingredients: Preserved wooly Mammoths, (with DNA hypothetically intact) and a close living relative. All we have to do is grab an Asian elephant egg, scoop out the DNA. Insert Mammoth DNA, and then insert the new embryo into an Asian Elephant womb. Bingo. Dolly 2 Wooly Mammoth Edition. One new cloned wooly mammoth, and even more awesome, it would come at a relatively cheap estimation of 10 million dollars. (According to this article, which I remember reading back in 2008 but was incredibly hard to find.) Regenerating a Mammoth for $10 Million - The New York Times (nytimes.com)

Only, there were a few problems. Those listed in the article, and many more.

First, ideal conditions are hardly, well, ideal. It turns out that ice, permafrost, etc are only so good at preserving DNA. Due to several factors I’m too lazy to research, the 6.8 million years potency of DNA is often replaced with a much less forgiving number 60,000 years. Less than 1/10th of that time. Any longer and you can almost guarantee that the DNA is so busted it has to be reconstructed.

And even that 60-thousand-year number is optimistic. As it turns out, the mammoths in the best condition, (frozen in ice or permafrost) are usually much older than the last mammoths to walk the earth. The mammoth baby sited last episode (blog) is about 30 thousand years old. It’s much older than the Wrangle Island Mammoths, but, still better preserved. This 30 thousand years and ‘better preserved’ is actually quite a hurdle.

To see what I mean, let’s talk about a method for bringing back mammoths which seems so obvious that, by rule of common sense, should have been mentioned before, right? Why don’t we just get some mammoth sperm, or eggs from frozen mammoths, and use those?

Well…

While it may seem to be the most obvious way to bring back Mammoths. It, actually, is not. We’d have to luck into viable mammoth sperm, and viable mammoth eggs. Which don’t exist. Those mammoths frozen in ice are dead. (I know, surprising right?) What does that mean? Well, it means that the cells which appear intact in photos haven’t actually been enacting biological processes for a long time. They are damaged by cold, they are damaged by thawing, and they don’t really work.

You need more than just the DNA for 2 gametes to make an embryo, you need the rest of those cells to be functional enough to help that DNA turn into an embryo.

So… we scoop that DNA out of those frozen mammoth gametes, then put them into new Asian elephant gametes and then use those to make an embryo. Several problems. At this time, there are no methods to replace the DNA inside a sperm, or an egg. (before fertilization. Introducing a nucleus after fertilization has been done.) But there’s an even bigger hurdle to this process. We’d need fully viable DNA from both gametes.

We’d probably need fully viable DNA from lots of gametes, actually. Dolly was the only viable embryo from a batch of 270 sheep embryos. (I know that’s cloning, which is different process from the imaginary gametes replacement method I’m dreaming up, but since my hypothetical process is even more experimental, then that probably means it will have an even larger failure rate. And we’re talking about failures involving for 4 gametes every time. (mammoth sperm, Asian elephant sperm, mammoth eggs, and Asian elephant eggs.)

Also, Viable DNA. That’s actually a sticking point too. Because DNA which has been frozen in ice for 30-thousand-years is never 100% intact. In ideal situations ½ of it degrades every 521 years. Bonds breaking, chromosomes shattering, nucleotides nucleotising. We’d be incredibly lucky to find 1 intact chromosome inside a gamete, much less a whole ½ mammoth’s worth. The chances of finding an intact sperm or egg nucleuo’s with all the DNA still together is smaller than winning the lottery. And we’d have to repeat this lottery level of luck thousands of times, invent a process to combine those lucky mammoth chromosomes into a single nucleus and then put that nucleus into new gametes.

Basically, this process would be even more confusing and labor intensive than reading the last four paragraphs.

While this mammoth-cicle may look like a prime sources of Mammoth DNA, unfortunately, time and freezer burn have rendered most of those genetic codes illegible.

So that brings us: Back to cloning. Let’s talk the Dolly Method. Its much easier. It wont work for Mammoths. We get DNA from an Adult Mammoth cell, then we put that into the nucleus of an unfertilized Asian Elephant Egg, and then Bob’s your uncle, you have that mammoth clone I mentioned earlier. Dolly 2 Mammoth Flavor. Except. Viable DNA is still a problem. It’s not like adult Mammoth Cells are any better at holding up to time. DNA falls apart. It gets contaminated with bacterial DNA. Again, finding a fully viable adult mammoth nucleus is going to be nearly impossible.

Then we’d still have to successfully transplant that nucleus to an egg and get the new cell to remain viable long enough to be implanted into an elephant. Lather rinse and repeat and at an incredibly optimistic failure rate of 1:270, that one lottery winning intact adult mammoth nucleus is gone and we just lost our best sequence of mammoth DNA.

Time to win the lottery over and over, and over and over.

Cloning a Mammoth in the method we cloned Dolly is not very likely. This is not an original thought.  Here’s an article about it written way back in 2013. Produce mammoth stem cells, says creator of Dolly the sheep (theconversation.com).

Ok. So the method they hit upon in this article actually provides another potential avenue for bringing back mammoths. Since we aren’t operating with a still living supply of adult mammoth cells, DNA intact, what if we get a single adult mammoth cell, turn it into a stem cell and clone that, over and over again to get lots of mammoth nuclei? Then we use these to fertilize an Asian Elephant embryo.

Sure. Assuming we find a viable adult mammoth cell. Which is still quite a hurdle, and even then, what do we get at the end of this process. Well, assuming we are able to successfully harvest an intact adult mammoth nucleus, transfer it into an Asian elephant cell, convert that into a stem cell, clone it enough times that we don’t have to worry about losing our intact mammoth nucleus anymore, implant it successfully into an Asian elephant egg, (repeat at least 270 times) and implant that into an elephant and raise it to term we get… 1 genetically viable Mammoth, with shortened telomeres (a problem with dolly style adult cloning which may or may not have a negative impact on clones.)

1 genetically viable individual does not a population of mammoths make. It can turn into a couple of half mammoths in a generation, and several more quarter mammoths in 2 generations, but we slowly find ourselves having done all of that work only to slide either towards massive genetic sustainability problems, or our mammoth family quickly becomes Asian elephants who can trumpet about their one hairy ancestor, and their ability to slightly better tolerate the cold. Elephants who can wear shorts in the winter.

Let’s say we find a couple of intact adult mammoth cells, (from different mammoths, otherwise we just run into the same problem but with slightly bigger numbers,) This is still becoming a labor intensive process with a high rate of failure, and that initial discovery of intact adult mammoth nuclei is the single most fragile point of failure in this plan. Is it possible? Sure, why not. I’d bet $1 that there’s an intact mammoth nucleous out there somewhere which can be clone, but I’d bet $100 dollars we’ll never find it. Let’s put Mammoth Stem cell cloning into the Way too much effort, for almost no reward file.

But this article from 2013 brings with it a whole new era of de-extinction discussion.

It’s the 2010’s baby and we don’t need viable nuclei anymore. We don’t even need intact chromosomes. All we need is a gene sequencer, a genetic map, and a fun little editing tool called CRISPR.

Oops…. I was going to get into the modern plans to bring back Mammoths this time, but instead I had way too much fun making seemingly viable plans seem as ridiculously complicated as they turn out to be.

Our biggest problem bringing back any of these old animals is that we do not have intact DNA. Dinosaurs, we’d be lucky to find nucleotides. Dodo’s we’d be fortunate to find relatively intact cells. Thylacines ditto, and mammoths the best preserved, with the most distinct specimens of the group lay in even worse state because of the ravages of time (and freezer burn.)

So have we come to a stumbling block yet? No. Because the present is often the key to the past, and as it turns out the key to the Mammoths might lie much more directly in their presently alive (ie: Not yet extinct) relatives the Asian Elephant. Or maybe even the African Elephant, Why not? At this point I’m willing to throw a mouse into the picture too. (because someone else did in an article I will reference next time.)

Why, a mouse?

How will we bring Mammoth’s Back?

Check out the next episode.

Also, Mammoths. Mammoth Cave is a place that has nothing to do with them, but I did write a book about it. Check it out. The best way to support me, and some awesome places, is to buy The Specters of Mammoth Cave from the stores which carry it. Hidden River Cave, Diamond Caverns, or the Mammoth Cave Book Story, (and hopefully, by the time you are reading this Barnes and Nobel in Bowling Green Ky.) But if you are nowhere near any of those places, you can also support me directly, and (for a limited time get 0.99cent shipping if you buy my book from this link to Ingram Spark: Buy The Specters of Mammoth Cave (ingramspark.com)