Deformed Animals Highlight Risk Of Gene Editing Technology

Deformed piglet resulted from Chinese efforts to resist viral infection. (From WSJ, Dec. 15-16, p. C3)

Crispr - let's reference - stands for "clustered regularly interspaced short palindromic repeats" - serves as the immune system of bacteria, and has been used to fend off viral attacks.  RNA or ribonucleic acid is a molecule found in all cells.  Like DNA, RNA is a polymer, but much simpler. This is because the latter’s ribose sugar molecules are smaller than DNA’s deoxyribose sugar molecules. So there are fewer monomer units in all to make polymer chains. At the same time, while there is only one type of DNA, there are three types of RNA:

- Messenger RNA or mRNA: really a complementary copy of a DNA segment that conveys information from cell nucleus to cytoplasm. Once there it acts as a template for synthesis of a protein molecule.

- Transfer RNA or tRNA: as the name implies, it couples information in the nucleic acid base sequence to the amino acid base sequence in synthesized proteins.

- Ribosomal RNA or rRNA: forms the ribosomes of cells, from which proteins are made.

The diagram below summarizes the process:






Back in October, 2017,  we learned that scientists had created a new Crispr- based system to edit RNA instead of DNA in human cells,. This offered  a way around some of the ethical and scientific challenges to do with editing the human genome.  This was first reported in the popular press  (WSJ Technology section, Oct. 26, 2017) and referenced a paper published in Science the day before in which the research was described. The new RNA -editing system was dubbed "Repair" by the researchers (based at the Broad Institute of MIT and Harvard)  and was designed to correct a common mutation known to play a role in a number of diseases.

This came on the heels of  attempts by Junjiu Huang  and colleagues to use CRISPR/Cas9 to fix a gene known as HBB, which causes  the blood disorder beta-thalassemia.   The work was done on 86 very early embryos that weren't viable, in order to minimize some of the ethical concerns. Only 71 of the embryos survived, and just 28 were successfully edited. But the process also frequently created unintended mutations in the embryos' DNA. 


Subsequently, Marcy Darnovsky of the Center for Genetics and Society, a watchdog group, wrote in an email :

"This paper demonstrates the enormous safety risks that any such attempt would entail, and underlines the urgency of working to forestall other such efforts. The social dangers of creating genetically modified human beings cannot be overstated. No researcher should have the moral warrant to flout the globally widespread policy agreement against modifying the human germline,"

George Daley, a stem cell researcher at Harvard, agreed, asserting:

"Their data reinforces the wisdom of the calls for a moratorium on any clinical practice of embryo gene editing, because current methods are too inefficient and unsafe," he wrote in an email. "Further, there needs to be careful consideration not only of the safety but also of the social and ethical implications of applying this technology to alter our germ lines."   

These concerns have not been proffered without justification, as we've since learned of other misfires in the gene editing arena  - most of which concern deformed animals unexpectedly generated by the work. (cf. 'Deformities Shadow Gene-Editing Research', WSJ, Dec, 15-16)

What were some of these misfires? As reported in the WSJ piece:

- When Chinese researchers deleted a gene that limits muscle growth in mammals the creations exhibited enlarged tongues.

- Similar experiments on Chinese pigs led a number of them to develop  additional vertebrae.

- Gene-edited calves died prematurely in Brazil and New Zealand. 

- Merino sheep edited to try to generate new coat colors came out spotted like typical Pandas.

In the case of the deformed piglet highlighted in the image the Chinese scientist - Dr. Kui Li-  didn't even use CRISPR (preferred because it's more economical than older techniques) but an older editing method. That involved deleting a gene called MSTN from Chinese pigs in the embryonic stage. The edited cells were then infused into eggs, chemically fertilized in a lab an implanted into the womb of a surrogate.

The experiment did produce leaner pork (up to 12 percent leaner if both copies of the MSTN gene were deleted) but also 1 in 5 pigs that developed an extra thoracic vertebra.  According to Dr. Li, lab tests showed that the meat of the deformed animals was still safe to eat. Maybe, but I'd warrant you'd have a hard time convincing my Bajan biologist friend John Phillips of that! E.g.  

 He will not even touch any genetically modified plant-based foods especially those which include glyphosate.  John still cites a particular paragraph in the Bulletin of the Food & Water Watch, from 2016:

"Very little is known of the long term effects of what GMOs can do to your health because they've not been properly tested. But what facts we do have are alarming. Real lab rats fed GMOs have suffered kidney and liver damage, cancerous tumors and even premature death."

So there's no doubt he wouldn't look twice at genetically modified pork, especially coming from deformed animals.

This then goes to the other effort, conducted at Nanjing Agricultural University - to use CRISPR to edit MSTN out of rabbits to make them meatier.   The unexpected result? 14 of the 34 engineered offspring were born with enlarged tongues. Would John Phillips eat any of these?  "No way, not now or ever!"

The bizarre result led the participating scientists to warn of "abnormalities"  from gene editing in a 2016 research paper on their project as cited in the WSJ piece. As the authors wrote:

"Safety issues need to be addressed in future studies before the technology can be utilized."

To be sure, these Chinese scientists weren't worried about humans getting enlarged tongues if they ate the rabbits. But rather the potential effects from consuming meat from an animal that had been "genetically messed up" to use Mr. Phillip's parlance.  As Phillips put it in a recent email: 

"Would you eat a flying fish I produced from Crispr/Cas9 gene editing that had two heads, two stomachs, and no scales?  How about just one head, but no dorsal fin and crossed eyes? No, I didn't think so! So why eat pork from a genetically-edited  deformed pig, never mind it's just one additional vertebra?"

Still, the beat goes on, and the WSJ article reports that globally "at least a dozen gene -edited livestock projects are aiming to reach consumer markets."

There is also a very good chance we won't know exactly what the condition of the animals was (in terms of deformities)  when their meat is  offered for the markets. After all, in the piglet experiment that went awry the Chinese scientist (Dr. Li) insisted it "was safe to eat".   But what does that mean? Safe in terms of not causing kidney or liver disease, say like glyphosates in GMO foods?  What about the precautionary principle? Is that embraced? (I.e. it is the responsibility of the creators to prove their meat is safe, not for consumers to prove it isn't.)


So I guess we should just  take his word for it,  being a geneticist and all.  As for me, I believe I will stick to the non-gene edited meats,  originating from non-deformed pigs, chickens, beef etc.  - which I am sure will disappoint the Neoliberal market mavens. The same lot who assert Americans need to have many more babies, never mind they can't afford them.

Appreciating The Dangers Of "Benevolent" Gene Editing Of DNA Using Crispr

Back in October we learned that scientists had created a new Crispr- based system to edit RNA instead of DNA in human cells offering a way around some of the ethical and scientific challenges to do with editing the genome.  This was first reported in the WSJ Technology section on Oct. 26 this year and referenced a paper published in Science the day before in which the research was described. The new RNA editing system was dubbed "Repair" by the researchers (based at the Broad Institute of MIT and Harvard)  and was designed to correct a common mutation known to play a role in a number of diseases.

Crispr - let's reference - stands for "clustered regularly interspaced short palindromic repeats" - serves as the immune system of bacteria, and has been used to fend off viral attacks.  RNA or ribonucleic acid is a molecule found in all cells.

Like DNA, RNA is a polymer, but much simpler. This is because the latter’s ribose sugar molecules are smaller than DNA’s deoxyribose sugar molecules. So there are fewer monomer units in all to make polymer chains. At the same time, while there is only one type of DNA, there are three types of RNA:

- Messenger RNA or mRNA: really a complementary copy of a DNA segment that conveys information from cell nucleus to cytoplasm. Once there it acts as a template for synthesis of a protein molecule.

- Transfer RNA or tRNA: as the name implies, it couples information in the nucleic acid base sequence to the amino acid base sequence in synthesized proteins.

- Ribosomal RNA or rRNA: forms the ribosomes of cells, from which proteins are made.

The diagram below summarizes the process:











Bio-Scientists, while they've long believed RNA to be a good therapeutic target, have actually focused on a less known enzyme -  Cas9 - for making edits in DNA, in their initial work with the Crispr system.

Let's note here RNA editing in general carries fewer potential scientific risks and - unlike DNA, cells constantly produce more RNA.  What kind of risks are we referring to in respect of DNA editing?  Well, not too long ago, Junjiu Huang  and his team of researchers attempted to make use of
a new technique called CRISPR/Cas9, which many scientists are excited about it because it makes it much easier to edit DNA.  In their process the Chinese team tried using CRISPR/Cas9 to fix a gene known as the HBB gene, which causes  the blood disorder beta-thalassemia.   The work was done on 86 very early embryos that weren't viable, in order to minimize some of the ethical concerns. Only 71 of the embryos survived, and just 28 were successfully edited. But the process also frequently created unintended mutations in the embryos' DNA. 

The worrisome aspect of the research is that  some of those mutations are critical to development and survival.   The report, in the journal Protein & Cell, was immediately condemned by other scientists and watchdog groups, who argued the research was unsafe, premature and raised disturbing ethical concerns.

Subsequently, Marcy Darnovsky of the Center for Genetics and Society, a watchdog group, wrote in an email :

"This paper demonstrates the enormous safety risks that any such attempt would entail, and underlines the urgency of working to forestall other such efforts. The social dangers of creating genetically modified human beings cannot be overstated. No researcher should have the moral warrant to flout the globally widespread policy agreement against modifying the human germline,"

George Daley, a stem cell researcher at Harvard, agreed, asserting:

"Their data reinforces the wisdom of the calls for a moratorium on any clinical practice of embryo gene editing, because current methods are too inefficient and unsafe," he wrote in an email. "Further, there needs to be careful consideration not only of the safety but also of the social and ethical implications of applying this technology to alter our germ lines."

Scientists have been able to manipulate DNA for years. But it's long been considered taboo to make changes in the DNA in a human egg, sperm or embryo because those changes could become a permanent part of the human genetic blueprint. One concern is that it would be unsafe: Scientists could make a mistake, which could introduce a new disease that would be passed down for generations. And there's also fears it this could lead to socially troubling developments, such as "designer babies," in which parents can pick and choose the traits of their children.

All of this in tandem explains the current switch to RNA editing - given that if "off target" changes occur the RNA edits are potentially reversible.  RNA editing also allows researchers to revise the therapy more easily as more research is done.

In the words of Elizabeth McNally, Director of the Center for Genetic Medicine at Northwester University's Feinberg School of Medicine:

"Crispr editing of RNA creates more opportunities for things we can do therapeutically"

But let no one think this editing is easy. Omar Abudayyeh, one of the co-authors of the Science paper,  noted the research team spent more than 18 months studying different Cas13 enzymes  to find the most effective to adapt for RNA editing in human cells.  (In nature there are many different Cas13 enzymes that bacteria use to target and cut RNA as art of their defense system.)

For her own part, Dr. McNally is hard at work on efforts to use the Crispr system to treat forms of muscular dystrophy. Meanwhile in September, Alnylam Pharmaceuticals reported positive results in a trial of a so-called RNA interference drug called patisiran. This drug stops production of a disease -causing protein to treat a rare nerve disorder: familial amyloid polyneuropathy.

There are new treatment fronts ready to open with the Crispr technique, we just have to be patient. Alnylam spent 15 years developing its RNA editing therapy.

Asteroids: May Not Be Exclusive Life Destroyers - But Also 'Creators'

Recent experiments published in the Proceedings of the National Academy of Sciences evidently show that asteroids may not be only destroyers of life, but also "creators" - well, ok, triggers for its later development. Basically, scientists in a lab used a powerful (1 billion kw)  laser to re-create what could have been the primordial spark of life on Earth.

They zapped a mix of clay and a chemical soup with the laser to simulate the energy of an incoming asteroid which usually enters the atmosphere of a speeds of 11,000 m/s.  to 30,000 m/s. The result? Crucial building blocks of life emerged - which bears similarity to the Miller-Urey experiment.

Recall the Miller and Urey experiment basically applied an electrical discharge to a chemical brew resembling the Earth’s primitive reducing atmosphere. This brew included ammonia and methane, as well as hydrogen and water vapor. The effect of the discharge transformed the mix into a diverse yield of organic compounds. These included amino acids, as well as substances such as formic acid and urea that normally occur in living organisms. 

In the latest study, the researchers concluded:

"These findings suggest that the emergence of terrestrial life is not the result of an accident but a direct consequence of the conditions on the primordial Earth and its surroundings."

Is that extravagant? Well, the experiment produced all four chemical bases: A (adenine), C (cytosine), G (guanine), and T (thymine), to make RNA, a simpler relative of DNA which is the blueprint for all terrestrial life. Like DNA, RNA is a polymer, but much simpler. This is because the latter’s ribose sugar molecules are smaller than DNA’s deoxyribose sugar molecules. So there are fewer monomer units in all to make polymer chains

What is notable about this laser experiment is that it's the first to test the theory that the energy from an asteroid impact could trigger the crucial chemical reactions.. By contrast, the Miller-Urey experiment presumed monstrous electrical discharges say from lightning strikes - to incept the amino acids in the primitive Earth.

According to lead author, Svatopluk Civis, his team used a laser almost 500 ft. long which operated for only a fraction of a second in order to impart the chemical soup with the requisite energy.  Civis averred that the power was so intense and concentrated that for less than a billionth of a second it equaled the output from "a couple of nuclear power plants".

Specifically, the laser beam produced roughly one billion kilowatts over a fraction of an inch generating heat of more than 7,600 F. The energy equivalent would be mind boggling and interested readers can work it out for themselves using the equation relating power to energy and time: E = P/t.

(Bear in mind 1 watt = 1 joule/ second or 1 J/s). The time t, meanwhile, is a billionth of a second or t = 0.000000001 s.

Why use an experiment based on an asteroid impact? Basically, because the emergence of the earliest life forms on Earth coincided with  period called the 'Late Heavy Bombardment' - when the asteroid belt was significantly larger and asteroids struck the Earth more often.  This is according to the study's co-author, David Nesvorny, of the Southwest Research Institute in Boulder, Colorado.  He notes that in the epoch referred to asteroids bombarded Earth ten times more frequently than before or after.

Of course, there are also the inevitable naysayers. For example, one researcher - John Sutherland - of the MRC Laboratory of Molecular Biology in Cambridge, England, insisted the amount produced of one base was so small that "the results don't seem relevant".

But Steve Benner, a prominent biological chemist, has maintained that the experiment is quite relevant because it produced the starting material that would have been around in the early Earth - irrespective of amount.

As standard for scientific practice, we will have to await further confirmation of the laser experiment to see if similar results can be obtained - or even enhanced.