CRISPR-Cas9

How a Rogue Chinese Experiment Might Affect Gene-Based Therapies in India

Many Indian scientists edit the human genome in cells obtained from their patients.

The gene-editing tool CRISPR Cas9 became a talking point among biologists after He Jiankui, a Chinese researcher, announced he had edited the genomes of two babies in November. In a YouTube video, Jiankui explained he had cut the CCR5 gene out to make them resistant to HIV. The episode set off a furore in the international biologists community.

The genetic editing of human embryos is banned in most of the world. This is partly because scientists are still learning how to use CRISPR to do this.

“CRISPR has certainly made gene-editing easier, but the strategy is not entirely foolproof and may sometimes introduce mistakes at unintended positions” of the genome, Debojyoti Chakraborty, a scientist at the Institute of Genomics and Integrative Biology, New Delhi, told The Wire. What these off-target changes could cause remains uncharted territory.

Sonam Mehrotra, a scientist at the Advanced Centre for Treatment, Research and Education in Cancer, Mumbai, said, “From practical experience, I can tell you that the outcome of CRISPR varies between cases.” Mehrotra uses CRISPR to deliberately introduce mutations in fruit flies (Drosophila melanogaster). Given the vagaries of working with flies, whose genomes are much smaller, she thinks it’s way too early to edit human embryos.

However, both Mehrotra and Chakraborty agree CRISPR has a lot of potential to treat genetic disorders.

Also read: Editing Embryos – Six Steps to an Informed Opinion

The human genome is a long sentence composed of four alphabets: A, T, G and C. Sometimes, when a wrong alphabet appears in a given position in the sentence, it could cause a genetic disease. Armed with CRISPR, scientists are learning how they can correct these mistakes by editing the faulty part.

This is the opposite of what Jiankui did. The embryos he edited didn’t have any genetic errors that made them susceptible to HIV. Instead, Jiankui modified the sequence of letters such that the newborn twins were “ resistant to HIV,” Chakraborty explained. ‘This kind of preventive medicine is not what scientists are aiming for, especially now, while we are still trying to understand CRISPR biology.”

This rather questionable use of CRISPR has since sparked a dialogue on human gene-editing regulations.

Tinkering with genetic material is a sensitive issue because it impinges on one’s identity. At present, there is an international moratorium on the genetic editing of human embryos for reproductive purposes. The practice has even denounced by China’s medical board; it only permits the editing of human embryos less than 14 days old.

In India, the ethical guidelines of the Indian Council of Medical Research (ICMR) disallow any research related to germline genetic engineering or reproductive cloning. But editing the genomes of adult human cells is permissible subject to approval by an ethics committee.

In administrative terms, “we have three levels of regulation,” S.R Rao, senior advisor to the Department of Biotechnology, said at a talk at the second International Summit on Human Genome Editing. It was held in Hong Kong in the last week of November 2018, and counted Jiankui among its participants.

At the first level, an institutional ethical committee screens all proposed projects. The approved ones are then assessed by a risk evaluation body of the Department of Biotechnology. In cases where there could be risks to the environment, the project may require an ‘okay’ from the environment ministry as well.

All clinical trials involving genetic engineering products come under the Drugs and Cosmetics Act, 1940.

Thus far, this labyrinthine process has not deterred scientists from exploring CRISPR as a treatment option for genetic disorders. Many Indian scientists edit the human genome in cells obtained from their patients. Their biggest focus area is developing a therapy for blood disorders due to defects in single genes.

This form of genetic editing is distinct from embryonic editing because it involves adult patient cells, not germ cells. The problem with editing the DNA of embryos is that both somatic and germ cells will carry the mutation, and the edited DNA will be passed on hereditarily.

But when working with adult cells – such as blood cells from patients – CRISPR can be used to correct mutations in the lab. And since the technique is patient-specific, any alterations made in the genome remain confined to one individual and won’t affect the next generation.

In that sense, CRISPR presents a lot of opportunities. China realised this early and eased regulations. And so far, Chinese scientists have used CRISPR to edit the genes of monkeys, human embryos less than 14 days old and are currently testing CRISPR-based therapies in cancer patients.

Things are moving in the US as well. American scientists having embarked on CRISPR-related clinical trials, with the country’s Food and Drug Administration keeping a close watch.

India is yet to start.

At the moment, scientists like Chakraborty are testing proof-of-concept studies in the lab, where he and his colleagues attempt to correct mutations in human blood cells. The next step is to test the technique in animal models, and then begin human trials. The timeline hasn’t been finalised, however.

“Technically, genome editing is not a challenge,” Chakraborty said. But one of CRISPR’s bigger caveats is that one size doesn’t fit all. “The outcome of CRISPR-based therapies will differ from one cell to another, from one gene to another and even from one delivery agent to another,” Chakraborty said. So each clinical trial will have to seek approval on a case-by-case basis. This does suggest officials will pay each proposal the attention it deserves – but it still also falls short.

Also read: Is There More to Gene Editing Than Creating ‘Designer’ Humans?

What India really needs, and lacks at the moment, is a proper framework to regulate CRISPR-based clinical trials. There has been a lot of talk about the need for new policies “but India is unlikely to go the China way in hurrying things up,” Deepti Trivedi, a scientific officer at the National Centre for Biological Sciences, Bengaluru, told The Wire.

Mehrotra is particularly worried about how Jiankui’s brazen foot forward may affect the future of CRISPR Cas9. “If a handful of researchers continue these reckless acts, governments may impose stringent regulations on the use of CRISPR.” This has already happened with stem cells: fearing rampant malpractice, the ICMR banned the use of stem cell technology in 2017 and limited access to them.

Jiankui’s actions prompted a sharp response from the Chinese government. It launched an investigation into his work, his university suspended him and he might be under house arrest. At the same time, scientists rallied to plan a gene-editing meeting in Massachusetts this year, where social scientists and geneticists will discuss the technology’s central issues.

Notwithstanding whether Indian scientists are going to be part of this conversation, Chakraborty believes something similar – “a dialogue between scientists and policymakers” – needs to happen in India. If it doesn’t, this path-breaking tech might never leave the pharmaceutical fringes it currently occupies.

Sarah Iqbal is a freelance science writer.

Chinese Geneticist Says Another ‘Potential’ Gene-Edited Pregnancy

He Jiankui, who has been challenged by several peers, says he is proud of his work.

Hong Kong: A Chinese scientist at the centre of an ethical storm over what he claims are the world’s first genetically edited babies said on Wednesday he is proud of his work and revealed that another there was a second “potential” pregnancy as part of the research.

He Jiankui, an associate professor at Southern University of Science and Technology in Shenzhen, China, addressed a packed hall of around 700 people attending the Human Genome Editing Summit at the University of Hong Kong.

“For this case, I feel proud. I feel proudest,” He said, when challenged by several peers at the conference.

“This study has been submitted to a scientific journal for review,” He said. He did not name the journal and said his university was unaware of his study.

He, who said his work was self-funded, shrugged off concerns that the research was conducted in secrecy, explaining that he had engaged the scientific community over the past three years.

In videos posted online this week, He said he used a gene-editing technology known as CRISPR-Cas9 to alter the embryonic genes of twin girls born this month.

He said gene editing would help protect the girls from infection with HIV, the virus that causes AIDS.

But scientists and the Chinese government have denounced the work that He said he carried out, and a hospital linked to his research suggested its ethical approval had been forged.

The conference moderator, Robin Lovell-Badge, said the summit organisers were unaware of the story until it broke this week.

Also read: The Exhilarating World of Neurogenetics

CRISPR-Cas9 is a technology that allows scientists to essentially cut and paste DNA, raising hope of genetic fixes for disease. However, there are concerns about safety and ethics.

The Chinese Society for Cell Biology in a statement on Tuesday strongly condemned any application of gene editing on human embryos for reproductive purposes and said that it was against the law and medical ethics of China.

More than 100 scientists, most in China, said in an open letter on Tuesday the use of CRISPR-Cas9 technology to edit the genes of human embryos was dangerous and unjustified. “Pandora’s box has been opened,” they said.

He’s research focuses on genome sequencing technology, bioinformatics and genome editing, according to his biography on the summit’s website.

He received his PhD at Rice University in Houston, Texas, and worked as a postdoctoral research fellow in Stephen Quake lab at Stanford University according to the site.

Continued study

He, who said he was against gene enhancement, said eight couples were initially enrolled for his study while one dropped out. The criteria required the father to be HIV positive and the mother to be HIV negative.

David Baltimore, president emeritus, Robert Andrews Millikan Professor of Biology, spoke after He’s speech, saying it was irresponsible to have proceeded until safety issues were in order.

“I don’t think it has been a transparent process. Only found out about it after it happened and the children were born,” he said.

He Jiankui said his results could be used for millions with inherent diseases. He said he would monitor the two newborns for the next 18 years and hoped they would support continued monitoring thereafter.

Shenzhen Harmonicare Medical Holdings Limited, reported by media as being involved in He’s project, sought to distance itself by stating the hospital never participated in any operations relating to the gene-edited babies and no related delivery had taken place.

In a statement published to the Hong Kong stock exchange on Tuesday, the group said preliminary investigations indicated the signatures on the application form circulated on the internet are “suspected to have been forged, and no relevant meeting of the Medical Ethics Committee of the hospital in fact took place”.

Chinese Scientist Apologises for Leak of Baby Gene-Editing Results

He Jiankui, who has claimed to have helped make the world’s first gene-edited babies, said that his study has been submitted to a journal for review.

Hong Kong: A Chinese scientist at the centre of a controversy over what he claims are the world’s first genetically edited children apologised on Wednesday for the result being leaked unexpectedly as he detailed his findings at a conference in Hong Kong.

“First, I must apologise that this result was leaked unexpectedly. This study has been submitted to a scientific journal for review,” He said. He did not name the journal and said his university was unaware of his study.

Also read: CRISPR Gene Editing and How It Works

In videos posted online this week, He said he used a gene-editing technology known as CRISPR-Cas9 to alter the embryonic genes of twin girls born this month.

He defended the work, saying gene editing would help protect the girls from infection with HIV, the virus that causes AIDS.

But scientists and the Chinese government have denounced the work that He said he carried out, and a hospital linked to his research suggested its ethical approval had been forged.

The conference moderator, Robin Lovell-Badge, said the summit organisers were unaware of the story until it broke this week.

Also read: CRISPR Gets Golden Makeover That Could Improve Gene-Editing Tech

CRISPR-Cas9 is a technology that allows scientists to essentially cut and paste DNA, raising hope of genetic fixes for disease. However, there are concerns about safety and ethics.

He’s research focuses on genome sequencing technology, bioinformatics and genome editing, according to his biography on the summit’s website.

He received his PhD at Rice University in Houston, Texas, and worked as a postdoctoral research fellow in Stephen Quake lab at Stanford University according to the site.

Chinese Scientist Claims to Have Helped Make World’s First Gene-Edited Babies

He Jiankui, of the Southern University of Science and Technology, China, used CRISPR in an attempt to remove a gene from cells in a human embryo and confer resistance to HIV.

New Delhi: A researcher in China has claimed that twin girls born this month are the world’s first genetically edited babies – and he helped make them.

According to an Associated Press report, He Jiankui claims to have altered embryos for seven couples during the course of their fertility treatments, and one woman got pregnant. In al, the ‘altered’ embryos, He says he attempted to build in resistance to HIV. He was not trying to cure or prevent an inherited disease, AP reported.

Before speaking to AP, He had also made his research available online (without mentioning the live births) and begun an outreach campaign to find out how the public would respond to such gene editing, according to Technology Review.

Gene editing of this kind is illegal in several countries, including India and the US. According to AP, several scientists have denounced He’s claims – which are yet to be verified – and said he is conducting ‘human experimentation’. The Chinese researcher told AP that he has been practising his process on mice, monkey and human embryos in the lab for several years and has applied for patents.

“I feel a strong responsibility that it’s not just to make a first, but also make it an example,” He said to AP. “Society will decide what to do next.”

He used CRISPR, the revolutionary gene-editing tool, according to Technology Review. He and his team were attempting to remove a gene called CCR5.

The researcher did not reveal the identities of the couples who were involved in his study, saying they wished to remain anonymous. He reportedly contacted them through a Beijing-based AIDS advocacy group named Baihualin.

He studied in the US before returning to China and opened a lab at the Southern University of Science and Technology of China in Shenzhen, AP reported. The researcher also owns two genetics companies. An American scientist helped He with his study, according to the news agency. Michael Deem was He’s advisor at Rice University and is on the board of the two companies.

The specifics

According to AP, He conducted the gene editing during an in vitro fertilisation procedure. All the men in the study were HIV positive and the women were not.

The sperm was separated from the semen because semen can include the HIV virus. A single egg and single sperm were combined to create an embryo. CRISPR was then introduced into this embryo.

Three to five days later, according to He, a few cells from the embryo were removed and checked for editing. Couples were then given a choice about whether or not they wanted to use the edited embryo.

He has claimed that tests show one of the twins has both copies of the intended gene altered, while the other has one copy altered. According to him, there is no evidence of any harm to other genes.

The second twin – the one with only one copy of the intended gene altered – can still get HIV, according to AP. There are a few studies that suggest that her health might decline more slowly than otherwise if she does get it, though.

Scientists who reviewed the test results He gave AP were not convinced of his success – both in terms of whether the editing was actually done and the possibility of other harm.

George Church, a noted geneticist Harvard University, one of the only American scientists quoted who did not raise ethical objections to He’s experiment, told AP that it was almost like “not editing at all” if only some cells were altered.

He’s decision to use the second embryo knowing full well that the altering had not worked completely was also questioned. “In that child, there really was almost nothing to be gained in terms of protection against HIV and yet you’re exposing that child to all the unknown safety risks,” Kiran Musunuru, a gene-editing expert at the University of Pennsylvania, West Philadelphia, and editor of a genetics journal, told AP.

Due process

It is not clear whether He’s research was totally above ground, per AP. For one, he reported his clinical trial long after he had actually started work on it, and when the twins were either already born or about to be. According to China’s registry of clinical trials, He gave official notice only on November 8.

Another question scientists have raised is around the information participants were given about the purpose and possible risks of the project. According to AP, the consent forms called the trial an “AIDS vaccine development” programme – which it is not.

Deem has stood by He and said he was present when the couples consented to the experiment. However, according to AP, neither of the two researchers have any experience conducting clinical trials.

He has said that he will cover the insurance requirements of the twins, and also follow their growth for as long as necessary. According to AP, further pregnancies under the clinical trial are on hold until the results of the first trial are clearly visible.

“I believe this is going to help the families and their children,” He told AP. If it causes unwanted side effects or harm, “I would feel the same pain as they do and it’s going to be my own responsibility.”

The ethics

The ethical questions, which – according to one analysis – could shape CRISPR’s future in solving human health problems to the extent that economics will, take two major shapes.

First, should humans alter life? Because editing one’s genes will effectively alter the template from which the body is constructed. Second, we are not aware of all the consequences that edited genes will have on the body as it ages. Should we still use it because the technique could cure a debilitating illness in the baby?

Priorities could help policymakers and lawmakers make better decisions on both fronts, especially when also informed by cultural norms. For example, using CRISPR to eliminate mosquitoes that spread malaria could have large social and economic benefits for countries ravaged by this disease.

On the other hand, many first-world nations that don’t have to do deal with a large malaria burden might not feel so compelled to resort to this technique.

China itself intends to be a global leader in biotechnology, and is at this time presumably taking advantage of a restrictive research environment in the former leader, the United States. The Asian superpower has eased regulations to the point where its scientists are able to experiment with human embryos.

Researchers around the world do acknowledge that tighter controls will be necessary to keep scientists, and regulators, from getting carried away. A popular example is designer babies: where parents might ask doctors to use CRISPR to ensure their soon-to-be-born child comes to life with certain features built-in, so to speak, and certain others removed.

Such selection could not just interfere with the supposed natural order but – more importantly – lend itself to eugenic tendencies with severe consequences for society.

Could All Shapes Have Their Origin in a Universal Genetic Code of Form?

The renowned architect Haresh Lalvani is thinking of ways in which the convergence of synthetic biology and nanotechnology will enable us to shape our environment based on the fundamental laws of nature.

Haresh Lalvani, a professor at the Pratt School of Architecture, New York, and co-founder and director of its Center for Experimental Structures, has long wondered if the way of the DNA molecule, which encodes all living structures, was a way that human-made designs could follow. He was in India briefly to receive a lifetime achievement award from IIT Kharagpur. While passing through Mumbai on his way back, he agreed to speak with S Ananthanarayanan for The Wire about his work.

Lalvani is an internationally recognised ‘architect-morphologist, artist-inventor and design scientist’ who looks at architecture beyond buildings, to take in shapes and forms of all description. His creations, on display as permanent installations in New York, in museums and in exhibitions, show glimpses of new directions and a breakaway from uniformity, with the use of technology and mathematics, for discovery, invention and industrial production.

The images (below) show some of his creations, a sculpture, a set of columns and a set of fruit platters, fabricated out of laser-cut sheet metal. The sculpture in the first image is made of welded pieces of quarter-inch-thick stainless-steel plate, and is installed at a busy street corner in New York. The second picture is of the AlgoRhythm Columns, 8-12 feet tall in folded sheet metal. They are a series from which a set of four in titanium are at the Museum of Modern Art, New York. The third picture is of the Morphing Platters, made of laser-cut steel, which were shown at an exhibition in 2011 presented by Murray Moss. Computer-generated designs have been combined with a mix of computer-controlled fabrication and traditional forming methods.

Credit: SEED54 (leftmost) by Peter Tannenbaum (Pratt Institute); Morphing Platters (rightmost) by Franklin Getchell

The concept behind the fluid harmony and facility of production of these creations is one that was inspired by the way living things build. Living things are mass-customised, with economy in production, yet each one is harmonious, suits its purpose and is unique. Lalvani conceives of a morphological genome, similar to its biological counterpart, made of DNA and specifies the structure of living things, as a mathematical construct to specify a set of numbers that define a structure. And just as changes in the units along the length of the DNA change the organism, varying the parameters along the morphological genome would alter the form that they represent.

Like the well-known Cartesian coordinates x, y and z, which pinpoint a location in three dimensional space, Lalvani proposes that the many features of a shape or design can be represented by numbers in a multidimensional space – a many dimensional space, one that we cannot physically build, called a hyperspace. Each point in hyperspace would then represent a shape or a form. And a change from one form to another would correspond to a path from one point to another in the space, and the changes in the numbers along the axes of the space. The sequences of these numbers represent the genetic code of form. In simple terms, in the same way we identify a shade of colour by specifying the three numbers for intensity of the primary colours, red, green and blue, we can specify a shape or form by a set of numbers indicated by a location in the morphological hyperspace.

An architect by training, Lalvani brings in a palette of skills and knowledge to guide his thinking. Biology, genetics, mathematics, particularly number theory, geometry and topology, chemistry, the science of materials, computers are all part of his ken. “It was a project we did in the final semester of my undergraduate architecture studies that set me thinking of architecture in a wider sense,” he says. “The assignment, in fact, was no assignment – we were asked to think of our own topic and work on it as we liked. Being set free brought with it a huge responsibility.” And in Lalvani’s case, it brought growth, study and contemplation, and a life-long search.

Haresh Lalvani

As an undergraduate, thanks to the environment at IIT Kharagpur, Lalvani formed a strong connection with mathematics, engineering, chemistry and biology. At the time, he was in contact with Robert Le Ricolais, the French visionary considered the ‘father of space structures’ and who used mathematics and natural structures in his work. There were also others who shaped Lalvani’s thinking, such as the German biologist J.G. Helmcke, Frei Otto – well-known for tensile and membrane structures – and also the turn-of-the-century Catalonian, Antoni Gaudi.

Buckminster Fuller, under whom Lalvani did his doctoral work, was a powerful influence. As an undergraduate student, Lalvani had reconstructed Fuller’s models using matchsticks and marbles, and reading Robert Horne’s article on the similarities between Fuller’s domes and virus structures – the first connection between architecture and nano-design in nature – along with electron micrographs of single-celled organisms sent to him in stereo images by Helmcke, were watershed moments in his development.

In the early 1970s, when he was deeply immersed in the relationship between architecture and the natural world, he was invited to a conference by James Watson, the co-discoverer of the DNA structure. “It was during this period that an idea struck me. The seemingly infinite variety of forms we see in biology, manipulated by just four chemical alphabets of DNA, raised a fundamental question for me. Is there an equivalent for architecture and human-made designs… is there a universal genetic code of form?”

Shaping and making

Over the years, Lalvani has developed the idea to considerable sophistication. The principles of morphogenomics, or morphological informatics, deal with indexing the numbers-based morphological genome so that it could yield families of form, from which architecturally relevant designs can then be derived. Once this indexing is done, similarly classified physical constraints would need to be overlaid so that only practical structures are encoded.

And then, when the limits placed by the program, function, our senses, limits of understanding and cultural constraints are imposed, Lalvani, in his 2003 essay ‘Organic Approach to Architecture’, says that what is left is “a morphologically structured network of information … that permits digital manipulation of form in the design process. When tied to digital manufacturing, this enables mass customisation in industrial production.”

Lalvani describes the egg-shaped sculpture in the street corner in New York, called SEED54, as a tiling structure, a curved surface using geometric shapes like the work of the noted Dutch artist M.C. Escher. SEED54 is made of irregular hexagons welded together, with curving, non-repeating apertures. The structure was inspired by a class of surfaces discovered by the mathematician-physicist Roger Penrose, with whose work Lalvani was deeply involved in the early 1980s, while completing his doctoral work.

SEED54 by Haresh Lalvani. Credit: Peter Tannenbaum (Pratt Institute)

In the AlgoRhythm columns, single, continuous metal sheets were shaped into fluid forms by computer-controlled machines, into geometries that arose from mathematics. The process, which Lalvani patented in collaboration with the company Milgo-Bufkin (which makes all his metal works), allows structures to be modelled and fabricated with greater freedom and flexibility than with moulds and dies, and obviously of lower cost.

“Integration of shaping (morphology) and making (fabrication) into a seamless whole, like it is in nature,” says Lalvani. “Seams are at the cornerstone… Seams take on hyperbolic shapes … and the seams guide the sinuous bends,” says Lisa Iwamoto, a professor at University of California at Berkeley, whose work focuses on fabrication techniques for architecture, in a review published by the Princeton Architectural Press.

Lalvani has since been interested in new technologies that could create self-forming shapes, or structures that erect themselves rapidly using not joints and rivets but continuous surfaces shaped by forces like gravity, of their own weight. One application that interests him is emergency shelters for people displaced by floods and hurricanes.

The Morphing Fruit Platters were laser-cut from stainless steel according to a computer program that generated patterns based on the mathematical form, the Archimedean spiral, and parameterised by the distance between a pair of points. As and when this distance was varied, the program turned out an unending series of surreal patterns, each different from others, harmonious but not symmetric. As the laser cutter was controlled by the computer, each round of cutting needed no fresh die or template and it used the same inches of cut. “We would have liked it if the patrons at the Design Miami 2011 exhibition, where we launched this product, could have ordered a pattern they saw on the computer screen and collected the product at the checkout.”

“The computer scientist Alan Turing had suggested that patterns like a zebra’s stripes, or a leopard’s spots, were based on interacting chemicals,” he continues. “Stephen Wolfram proposed ‘cellular automata’ rules that relate combinations of adjacent black and white cells as a basis for such patterns. But in the Morphing Platters, we generated patterns based on the distance between a pair of points. This was different.”

Coming back to Lalvani’s vision of structures taking shape based on the “DNA of form”, DNA technology is being used to design better crops, food, drugs and biofuels. We also have artificial intelligence, where machines are self-learning to become better at tasks ranging from image recognition to operating driverless cars. Lalvani’s ultimate vision is to see the link between physical building processes, synthetic or organic, to the morphological genome, to create “artificial architecture.”

There is a long way to go, but when this happens, “form, space, material and process will become one”, as in nature. “Objects will begin to design and shape themselves.” In the meantime, he is busy refining aspects of this work, filling in gaps here and there, extending where possible, and thinking of ways in which the convergence of synthetic biology and nanotechnology will enable us shape and make things at larger scales so that they, in turn, will shape our environment based on the fundamental laws of nature.

S. Ananthanarayanan has been writing columns for Indian newspapers on developments in science and technology for lay-person readers, and blogs at simplescience.in.

Explained: US Court Settles Bitter Gene Editing Patent Case, Confusion Lingers

The Broad Institute has retained its patent on using CRISPR-Cas9 in eukaryotes, preserving the dozen-plus licenses it has issued for commercialisation of the technology.

On September 10, a US court settled an increasingly churlish patent dispute between two research institutions in the country, the University of California (UC) in Berkeley and the Broad Institute, Massachusetts, with great consequence for the commercial use of a powerful gene-editing technology called CRISPR-Cas9.

The dispute centred on CRISPR’s usability in two different kinds of biological lifeforms: prokaryotes and eukaryotes. UC had devised a way to edit the genetic makeup of prokaryotes using CRISPR in 2014. Broad followed a year later with a method to use CRISPR in eukaryotes. UC had subsequently contested the patentability of Broad’s method saying that it was a derivative of UC’s method and couldn’t be patented separately.

The court as well as the US patent office have disagreed and upheld Broad’s patent.

The following FAQ breaks the case down to its nuclear components and assesses the verdict’s implications and future courses of action.

What are CRISPR and CRISPR-Cas9?

CRISPR is a natural defensive mechanism that prokaryotes use to protect themselves against viruses. Prokaryotes are smaller, less complex lifeforms and include bacteria and archaea: they are unicellular, the cells lack a membrane as well as membrane-bound organelles. The bigger lifeforms are classified as eukaryotes, which are multicellular, whose cells have a membrane and the cells also contain membrane-bound organelles. Because of these and other differences (see here, p. 8), it wasn’t clear if a CRISPR system built for use in prokaryotes could be adapted by a person of reasonable skill for use in eukaryotes with a reasonable chance of success. This distinction is at the heart of the patents dispute.

CRISPR-Cas9 is essentially a technology that combines CRISPR with a protein called Cas9 to form a molecular tool. This tool can swim to a eukaryote’s DNA, pick out a specific section of the genes and snip it at both ends, removing it from the DNA sequence. Once the section is out, the DNA strand repairs itself to restore the genes. If the original section of genes was faulty (e.g. containing an undesirable mutation), then CRISPR-Cas9 can be used to remove it from the DNA and force the DNA strand to repair itself to a healthier version.

Researchers have already reported that they are close to using this technology to treat a debilitating condition called Duchenne muscular dystrophy.

Of course, there are other gene-editing technologies, including zing-finger nucleases and transcription activator-like effector nucleases, but CRISPR-Cas9 has proved to be more efficient, effective and easier to use. At the same time, a few concerns are starting to emerge about unintended side-effects.

What was the case timeline?

The UC team, led by Jennifer Doudna, published a paper in August 2012 describing how an RNA-based system called CRISPR-Cas9 could be used to edit DNA in prokaryotic cells. Editing DNA is a lucrative prospect – then and now – because it allows us in theory to modify the fundamental constitution of biological life, curing debilitating illnesses as much as modifying crops. And what the UC team had found, together with Emmanuelle Charpentier, then of the University of Umea in Sweden, was the first tool that could achieve this. After their paper was published, the UC team filed a patent with the US Patent and Trademarks Office (USPTO) for the use of CRISPR in prokaryotes.

While UC’s patent was pending, a team led by Feng Zhang from the Broad Institute, setup by the Massachusetts Institute of Technology and Harvard University, Boston, published a paper in 2013 and then built a CRISPR system in 2014 that could work in eukaryotes. Zhang and co. then filed for an expedited patent that was granted in 2017. At this point, UC complained to the USPTO that the Broad patent infringed on its own – that, effectively, Zhang et al’s work was not patentably distinct from Doudna et al’s work. UC’s own patent for CRISPR use in prokaryotes was granted in early 2018.

In late 2017, the Patent Trial and Appeal Board (PTAB) of the USPTO upheld the Broad patent, effectively stating that the DNA-editing technologies used in prokaryotes and eukaryotes were “patentably distinct”. Specifically, it had ruled that there was no interference-in-fact, i.e. that UC’s general description of use of CRISPR in biological systems could not have anticipated, under reasonable circumstances, Broad’s more specific CRISPR invention for use in eukaryotes. An interference-in-fact check is pegged on a so called ‘obviousness review’. A 1966 SCOTUS case defines four factors using which it can be undertaken:

(1) the scope and content of the prior art; (2) the differences between the claims and the prior art; (3) the level of ordinary skill in the art; and (4) objective considerations of non-obviousness

UC decided to appeal the PTAB’s verdict with the US Court of Appeals of the Federal Circuit (CAFC). The latter came to its decision on September 10, ruling in favour of the USPTO and upholding the Broad patent.

What happens next?

A lot of things. Let’s classify them as financial, academic, legal and administrative.

Financial – Where there’s a patent, there’s money. However, there’s more money for Broad than for UC because almost all application of the CRISPR technology will happen in eukaryotes, a domain that includes humans and plants. And because the Broad patent has been upheld, this effectively means the UC patent can apply only to prokaryotes and not to eukaryotes.

Public attitudes to this affirmation were partly reflected in the share values of three companies intent on commercialising CRISPR tech: Crispr Therapeutics AG (cofounded by Charpentier) and Intellia Therapeutics have licenses with UC and their shares fell by 5.3% and 2.5% respectively; Editas Medicine Inc., which has licenses with Broad, climbed by 6.8%.

A review in 2017 stated that although “CRISPR IP ownership is claimed by at least seven different parties”, the Broad patent could be a “blocking patent” because of its ancestral nature. This is one reason why the Broad Institute has already issued 13 licenses, more than any of the other patent-holders. In all, the review estimated that the American gene-editing industry will be worth $3.5 billion by 2019, with CRISPR propelling biotechnology to the status of “second highest funded sector in the United States”.

Academic – The contest between UC and Broad has only worsened the mutual, and deleterious, embitterment between the institutions. In 2015, Broad launched an acrimonious campaign to turn public opinion in its favour, which included attempts to rewrite the history of DNA-editing research and present Zhang’s achievement in stronger light. The possibly most damaging thing Broad did was to quote UC’s Doudna herself as having expressed frustration and doubt about whether a CRISPR system for use in bacteria could be adapted for use in eukaryotic cells.

These quotes were used in Broad’s filings for the patent dispute, undermining UC’s case. However, scientists have argued that science is almost never free of frustration and that Doudna was also right to express doubt because that’s what any good scientist would do: lead with the uncertainty until something to the contrary could be demonstrated. However, Broad effectively penalised Doudna for being a good scientist – an action that Michael Eisen, a biologist in Doudna’s department at UC, has said is rooted in universities being able to profit from patents created with taxpayer dollars.

Legal – It’s important to recognise what UC has actually lost here. UC appealed the PTAB verdict, bringing it to the CAFC, who in turn ruled that the PTAB had not wronged in its conclusion. The judge did not reevaluate the evidence and did not hear arguments from the two parties; no new evidence was presented. The court only affirmed that UC, in the eyes of the law, did not have grounds to contend the PTAB verdict. A salient portion from the judgment follows, where the judge writes that some parts of the CRISPR/Cas9 system as used in prokaryotes could have been adapted for use in eukaryotes but that that’s besides the point (emphasis added):

UC expended substantial time and effort to convince this court that substantial evidence supports the view it would like us to adopt, namely, that a person of ordinary skill would have had a reasonable expectation of success in implementing the CRISPR-Cas9 system in eukaryotes. There is certainly evidence in the record that could support this position. The prior art contained a number of techniques that had been used for adapting prokaryotic systems for use in eukaryotic cells, obstacles adopting other prokaryotic systems had been overcome, and Dr. Carroll suggested using those techniques to implement CRISPR-Cas9 in eukaryotes. We are, however, an appellate body. We do not reweigh the evidence. It is not our role to ask whether substantial evidence supports fact-findings not made by the Board, but instead whether such evidence supports the findings that were in fact made. Here, we conclude that it does.

Therefore, this is a judgment of the law, not a judgment of the science.

According to Jacob Sherkow, a professor at the New York Law School, UC can either petition the CAFC for a rehearing or appeal to the Supreme Court. Sherkow added that neither strategy is likely to work because he doesn’t think “this case presents any *novel* legal issues” (emphasis in the original). This means UC will likely return to the patent office and attempt to “salvage what they can from their patent application”.

There is also another problem. To quote Chemical and Engineering News,

… it recently became clear that another CRISPR scientist, Virginijus Šikšnys of Vilnius University [Lithuania], filed a patent for CRISPR/Cas9 just weeks before UC Berkeley filed its patent in 2012. While UC Berkeley and Broad were entangled in their dispute, the Šikšnys patent was approved and made public, meaning that USPTO can now hold the Šikšnys patent against UC Berkeley. “That has the potential to sink whatever is left from Berkeley’s patent application,” Sherkow says.

Administrative – This part is confusing. In the US, the USPTO upheld the Broad patent in February 2017. But in Europe, the European Patent Office (EPO) ruled in favour of Doudna and Charpentier in March 2017. So depending on the jurisdiction, companies that want to commercialise CRISPR technology (for eukaryotes) will have to work with UC in Europe and the Broad Institute in the US. At least one company, DowDuPont, which is using CRISPR to engineer corn and soybean crops to be cultivable without pesticides, has purchased licenses with both institutions.

The different judgments arise from one difference in how the EPO and the USPTO evaluate ‘no interference-in-fact’. According to a May 2017 report by Sherkow, “In Europe, one is entitled to a broad patent on a new technique, if it demonstrates an ‘inventive step’ over prior methods, even if there [is] no guarantee that it will work for all of its claimed applications.” In the US, on the other hand, each “claimed application” has to be demonstrated and is separately patentable if one application doesn’t follow obviously from the previous. The EPO decision is open to challenge and Broad is likely to use the opportunity to do so.

By the way, the country with the second-most patents related to CRISPR is China, after the US. Chinese research institutions and industry players have been focusing mostly on knockout mechanisms of CRISPR, which control how undesirable genes in a DNA sequence are removed. To quote at length from the 2017 review,

The Chinese government has been actively involved in gene-editing funding. The National Natural Science Foundation of China (NNSF), invested $3.5 million in over 40 CRISPR projects during 2015. Through the NNSF and the National Basic Research Program, the Chinese government has funded the first use of CRISPR for the modification of human embryos. Additionally, Shenzhen Jinjia Color Printing Group Co., a public company, has pledged $0.5 million to fund Sun Yat-sen University for studying CRISPR in embryos.

What do scientists say?

Scientists’ reactions are still coming in, although no consensus is likely to emerge soon. In the meantime, awards make for a reasonable proxy to determine what scientists think is laudable. On this count, Doudna and Charpentier are clear leaders. Since 2014, Doudna has won 20 awards (excluding one from UNESCO), Charpentier has won dozens and Zhang, nine (although must be noted that Doudna and Charpentier have been scientists for longer than Zhang has). Doudna, Charpentier and Šikšnys were also jointly awarded the 2018 Kavli Prize in Nanoscience.

CRISPR Gets Golden Makeover That Could Improve Gene-Editing Tech

While the new version of the tech has room for improvement, scientists say it opens a new way for safer, more accurately controlled delivery of gene-editing tools.

Credit: JohannesW/pixabay

Lakshmi Supriya is a freelance science writer based in Bengaluru.

Snip, snip. The defective genes in your body are cut and replaced with a normal gene, and just like that, genetic diseases can be treated. Now, the new technology of gene editing that has taken therapeutics by storm, called CRISPR, has moved another step forward. Researchers have found a new, more efficiency, way of delivering the different components needed for the editing.

The origin of CRISPR dates back to the early 1990s, when Francisco Mojica discovered that DNA fragments of some microbes called archaea had several copies of a roughly palindromic sequences of bases – the building blocks of DNA – that were different from the sequences that are generally found in archaea. Called clustered regularly interspaced palindromic repeats (CRISPR), he found such sequences in several species of bacteria over the next decade.

More research by several groups suggested that these sequences helped the bacteria fight against attacks by viruses or other foreign bodies by incorporating parts of the viral DNA into the CRISPR sequence. These foreign DNA are called spacers and they act as a memory bank, allowing the bacteria to recognise the virus as an invader when it next attacks. Another key component of the bacteria’s defence system, the Cas9 protein, is responsible for chopping up the DNA of the invader, and is guided to the right position on the DNA by a piece of RNA.

Inspired by this natural defence mechanism in bacteria, CRISPR was modified to serve as a tool to edit genome sequences. By changing the sequence of the guiding RNA, the Cas9 protein could be directed to any desired location on the DNA. Once the particular genome sequence has been cut, either the body’s natural DNA repair mechanisms will fix it by simply gluing back the ends – or a specific sequence can be supplied to fill in the gap. A simple cut and paste that has now edited the DNA.

Although the technology is the simplest yet for gene editing, delivering all the components – Cas9, the guide RNA and the repair DNA – together is a challenge.

Cas9 is a set of proteins produced by bacteria, and delivering them has been a challenge. They cannot be simply isolated and injected into the system. Typically, a particular type of virus – known as the adeno-associated virus – has been used as a delivery vehicle for this protein. The virus was chosen because most humans are immune to it, and modified such that it can produce the protein.

However, there are several disadvantages of using a virus. One of the biggest problems is the chance of editing the genome in the wrong place because the virus does not stop producing the protein once the correction has been done at the desired location. The greater the amount of protein in the system, the greater the chance is that it will cut DNA, even if it is not at the right location. In addition, viruses are small, so a large number is required to produce sufficient editing. This issue becomes worse for human therapies because numerous copies of the virus will be required – often beyond clinically acceptable levels.

In a recent study, researchers found a way around this problem. Instead of using a virus, they used nanoparticles to deliver all the components simultaneously. “This study makes a major advance by demonstrating the delivery of all three [components], with promising results in human cells and in mice,” says Chase Beisel, a professor at North Carolina State University, who works on CRISPR systems and was not involved in the study.

In a method dubbed CRISPR-Gold, all three components can be attached around a gold nanoparticle and enclosed using a polymer, keeping it all together in one package. This package can be delivered into a variety of cells. Once it reaches a cell, the polymer breaks apart to release all the components.

“Gold is particularly easy to work with because of its facile reactivity with thiols,” says Niren Murthy, a professor of bioengineering at the University of California, Berkeley, and one of the authors of the study. Taking advantage of this fact, Murthy and colleagues designed DNA with thiol groups, which are molecules of sulphur and hydrogen. To this, they attached the repair DNA and the Cas9 proteins along with the guide RNA, ensuring that the nanoparticle now has all the components required for gene editing in a single package. Different types of cells easily take up gold nanoparticles, allowing easy delivery of the gene-editing agents.

“CRISPR-Gold and, more broadly, CRISPR-nanoparticles, open a new way for safer, accurately controlled delivery of gene-editing tools,” says Irina Conboy, a professor of bioengineering at the University of California, Berkeley, and one of the authors of the study, in a statement.

Using this system, researchers showed that they could edit genes in human embryonic stem cells and mouse muscle cells in the lab. In addition, they were able to remove a genetic mutation that causes a disease called Duchenne muscular dystrophy. This disease is a perfect candidate for the CRISPR technology because it is caused by a mutation in one specific gene, which prevents the body from producing dystrophin, a protein required for muscular development. It occurs in children and causes extreme muscular weakness. Starting with muscle loss in the legs, it progresses up the body. Within a few years, afflicted children cannot walk or stand, and may also suffer from intellectual disabilities. There is no known cure for the disease.

Injecting CRISPR-Gold into the muscles of mice afflicted with this disease significantly increased their muscle function almost twofold. Analysis showed that the mutated gene had been removed and that very low levels of untargeted DNA had been removed, about 0.005-0.2% for five different genomic DNA sequences tested.

However, the fraction of mutant DNA that was repaired is still very low, only about 0.8%. “While this frequency may be acceptable for diseases that can be reversed by editing a fraction of the cells, other diseases will require much more efficient means of editing,” says Beisel. Murthy speculates that about a 5% efficiency of repair is needed for therapeutic benefits. “We anticipate that we can achieve this by doing multiple injections,” he says.

Another concern is if the addition of a new system into the body would cause any adverse immune reactions. Testing the mice two weeks after injection showed that there had been no adverse immune reaction in them.

Although the system has effectively shown that non-viral delivery of the gene-editing components is possible, Murthy says they are working on improving it. One limitation of the method today is that it requires direct intramuscular injection to the diseased muscles. CRISPR-Gold can potentially be used for treating genetic diseases where tissue can be accessed with a direct injection. “However, developing formulations that can be injected intravenously would increase the types of diseases that could be treated with CRISPR-Gold,” he says.

Murthy also thinks that the size of the gold nanoparticles – about 100-200 nanometers wide – is too big for editing muscle after intravenous delivery. To bring the technology to market, Murthy and colleagues have formed a company called GenEdit and are now focusing on improving the rate of gene repair for muscular dystrophy.

However, Beisel imagines that the technology need not be limited to treating genetic diseases in humans. They could be used in gene therapy in other animals and plants, such as making disease-resistant plants or fatter animals for meat consumption. “Time will tell whether these nanoparticles would be effective for other cells, such as animals, plants, fungi or bacteria.”

The Future of Genetic Enhancement is in China, Not in the West

If the critics are correct that human enhancement is unethical, dangerous or both, then its emergence in China could be worrying.

Credit: tamakisono/Flickr, CC BY 2.0

Would you want to alter your future children’s genes to make them smarter, stronger or better-looking? As the state of the science brings prospects like these closer to reality, an international debate has been raging over the ethics of enhancing human capacities with biotechnologies such as so-called smart pills, brain implants and gene editing. This discussion has only intensified in the past year with the advent of the CRISPR-cas9 gene editing tool, which raises the specter of tinkering with our DNA to improve traits like intelligence, athleticism and even moral reasoning.

So are we on the brink of a brave new world of genetically enhanced humanity? Perhaps. And there’s an interesting wrinkle: It’s reasonable to believe that any seismic shift toward genetic enhancement will not be centered in Western countries like the U.S. or the U.K., where many modern technologies are pioneered. Instead, genetic enhancement is more likely to emerge out of China.

Attitudes toward enhancement

Numerous surveys among Western populations have found significant opposition to many forms of human enhancement. For example, a recent Pew study of 4,726 Americans found that most would not want to use a brain chip to improve their memory, and a plurality view such interventions as morally unacceptable.

A broader review of public opinion studies found significant opposition in countries like Germany, the U.S. and the U.K. to selecting the best embryos for implantation based on nonmedical traits like appearance or intelligence. There is even less support for editing genes directly to improve traits in so-called designer babies.

Opposition to enhancement, especially genetic enhancement, has several sources. The above-mentioned Pew poll found that safety is a big concern – in line with experts who say that tinkering with the human genome carries significant risks. These risks may be accepted when treating medical conditions, but less so for enhancing nonmedical traits like intelligence and appearance. At the same time, ethical objections often arise. Scientists can be seen as “playing God” and tampering with nature. There are also worries about inequality, creating a new generation of enhanced individuals who are heavily advantaged over others. Brave New World is a dystopia, after all.

However, those studies have focused on Western attitudes. There has been much less polling in non-Western countries. There is some evidence that in Japan there is similar opposition to enhancement as in the West. Other countries, such as China and India, are more positive toward enhancement. In China, this may be linked to more generally approving attitudes toward old-fashioned eugenics programs such as selective abortion of fetuses with severe genetic disorders, though more research is needed to fully explain the difference. This has led Darryl Macer of the Eubios Ethics Institute to posit that Asia will be at the forefront of expansion of human enhancement.

Restrictions on gene editing

In the meantime, the biggest barrier to genetic enhancement will be broader statutes banning gene editing. A recent study found bans on germline genetic modification – that is, those that are passed on to descendants – are in effect throughout Europe, Canada and Australia. China, India and other non-Western countries, however, have laxer regulatory regimes – restrictions, if they exist, are often in the form of guidelines rather than statutes.

The U.S. may appear to be an exception to this trend. It lacks legal restriction of gene editing; however, federal funding of germline gene editing research is prohibited. Because most geneticists rely on government grants for their research, this acts as a significant restriction on germline editing studies.

By contrast, it was Chinese government funding that led China to be the first to edit the genes of human embryos using the CRISPR-cas9 tool in 2015. China has also been leading the way in using CRISPR-cas9 for non-germline genetic modifications of human tissue cells for use in treatment of cancer patients.

There are, then, two primary factors contributing to emergence of genetic enhancement technologies – research to develop the technologies and popular opinion to support their deployment. In both areas, Western countries are well behind China.

Different countries have different expectations about working with human genes. Credit: Michael Dalder/Reuters

What makes China a probable petri dish

A further, more political factor may be at play. Western democracies are, by design, sensitive to popular opinion. Elected politicians will be less likely to fund controversial projects, and more likely to restrict them. By contrast, countries like China that lack direct democratic systems are thereby less sensitive to opinion, and officials can play an outsize role in shaping public opinion to align with government priorities. This would include residual opposition to human enhancement, even if it were present. International norms are arguably emerging against genetic enhancement, but in other arenas China has proven willing to reject international norms in order to promote its own interests.

Indeed, if we set ethical and safety objections aside, genetic enhancement has the potential to bring about significant national advantages. Even marginal increases in intelligence via gene editing could have significant effects on a nation’s economic growth. Certain genes could give some athletes an edge in intense international competitions. Other genes may have an effect on violent tendencies, suggesting genetic engineering could reduce crime rates.

Many of these potential benefits of enhancement are speculative, but as research advances they may move into the realm of reality. If further studies bear out the reliability of gene editing in improving such traits, China is well-poised to become a leader in the area of human enhancement.

Does this matter?

Aside from a preoccupation with being the best in everything, is there reason for Westerners to be concerned by the likelihood that genetic enhancement is apt to emerge out of China?

If the critics are correct that human enhancement is unethical, dangerous or both, then yes, emergence in China would be worrying. From this critical perspective, the Chinese people would be subject to an unethical and dangerous intervention – a cause for international concern. Given China’s human rights record in other areas, it is questionable whether international pressure would have much effect. In turn, enhancement of its population may make China more competitive on the world stage. An unenviable dilemma for opponents of enhancement could emerge – fail to enhance and fall behind, or enhance and suffer the moral and physical consequences.

Conversely, if one believes that human enhancement is actually desirable, this trend should be welcomed. As Western governments hem and haw, delaying development of potentially great advances for humanity, China leads the way forward. Their increased competitiveness, in turn, would pressure Western countries to relax restrictions and thereby allow humanity as a whole to progress – becoming healthier, more productive and generally capable.

Either way, this trend is an important development. We will see if it is sustained – public opinion in the U.S. and other countries could shift, or funding could dry up in China. But for now, it appears that China holds the future of genetic enhancement in its hands.

G. Owen Schaefer is Research Fellow in Biomedical Ethics, National University of Singapore.

This article was originally published on The Conversation.

Editing Embryos – Six Steps to an Informed Opinion

For the first time, a government has approved research involving modification of human embryos, but there’s a heavy context underlying this decision that needs to be considered before forming any kind of opinion.

For the first time, a government has approved research involving modification of human embryos, but there’s a heavy context underlying this decision that needs to be considered before forming any kind of opinion.

The UK’s permission for editing human embryos at the Francis Crick Institute doesn’t really mean designer babies. Credit: Wolfgang Moroder/Wikimedia Commons, CC BY-SA 3.0

Step 1: Understand why a scientist studies embryos

Hundreds of little physical, biological and chemical events have to take place before every human being is born. A sperm cell in the female reproductive tract has to overcome fatally acidic atmospheres, attacks from immune cells and multiple physical traps to finally reach the egg, then the nuclei of the egg and sperm must fuse correctly, and finally the resultant zygote cell must develop into a multicellular embryo and follow a fixed blueprint to successfully develop into a complete healthy human being.

Tiny mistakes in this chain of events suffice to give rise to problems like infertility, miscarriages or developmental disorders. One job of developmental biologists is to fully understand the genes without whose functioning healthy embryonic development would not take place.

Step 2: Understand why scientists modify genes

One of the most popular strategies to investigate the role of a gene in an organism is to create a “knock-out” version where that gene is inactivated and see what happens. Any deviation from normal behaviour indicates to the researcher what the function of the knocked-out gene could be.

The challenge with most traditional gene knock-out techniques is their efficacy. “While they work relatively well for simple organisms like bacteria and yeast, editing mammalian genes requires much more precision and control,” said Sunil Laxman, who uses gene modification techniques to study cell fate at the Institute for Stem Cell Biology and Regenerative Medicine (inStem) in Bengaluru. The number of cells that end up successfully incorporated with the desired alteration are far too few, and there is a very real risk of creating unintended mutations.

However, in 2012, a new technology called the CRISPR-Cas9 system burst into the scene and made gene editing easier, more efficient and quicker than ever before. “With CRISPR, the efficiency and performance of gene editing experiments, especially involving single point changes, becomes orders of magnitude better,” said Laxman.

Step 3: Understand the limitations of animal models

Since human embryos aren’t easy to get access to, owing to the ethical and safety implications of mutant humans, most developmental biologists have to make do with experimenting on model organisms like mice. “Mouse embryos are sufficient to answer some of our questions, but they’re not ideal for many others,” said Ramkumar Sambasivan, a developmental biologist at inStem.

For example, a 2015 paper titled ‘Only humans have human placenta’ concluded that the mouse’s is an inadequate representation of human placental development. “Many aspects of human placentation can only be understood on the basis of experiments on human cells and tissues in combination with data collections from human subject studies,” the authors of the paper wrote. In such scenarios, it becomes important for developmental biologists to seek human embryos to conduct research with.

Step 4: Understand why are wary of using human embryos

Studies involving genetic modification have so far mostly been limited to animal cells or non-reproductive human cells that pose no risk of passing on the mutation to future generations. This application is quite common and relatively non-risky, especially now with the advent of CRISPR-Cas9.

Experimentation on human embryos, on the other hand, is highly controversial. The fear is that embryonic research, owing to their heritability, is a slippery slope into the era of designer babies and unsafe and unapproved gene therapy rackets. But Sambasivan reminds us that this fear existed as long as gene modification existed. “Of course, CRISPR may have made this eventuality more imminent,” he acknowledges.

The specific regulations as well as the broader legal system around human embryonic research and gene modification are stringent yet blurry in most countries – since the nature, the technologies and implications of the research are so rapidly evolving. The US government, for example, will not fund research involving modified embryos.

The UK is a bit more relaxed. While it bans modifying embryos for clinical or therapeutic purposes, it leaves it up to its Human Fertilisation and Embryology Authority (HFEA) to approve modification of embryos for research. This is what allowed Kathy Niakan from the Francis Crick Institute to file her application to HFEA last October. And HFEA’s thumbs up earlier this month made this the first ever such governmental approval in the world.

In countries like Japan and India, the practice in restricted, but only by guidelines and not by law – and that’s neither here nor there. This blurriness, coupled with the large number of fertility clinics that exist in these two countries, prompted Nature to predict that India or Japan will be the first to produce live babies with edited genomes.

In fact, there is a dialogue that seems to be happening in India. At an international meet in December 2015, K. VijayRaghavan, the secretary of the Department of Biotechnology, said that a committee meeting discussing gene modification had taken place around the time and that the decision to organise public discourses had been taken. However, no more details were shared.

Step 5: Understand the motive of Kathy Niakan’s research

Once Niakan gets a final approval from one more ethics committee, she will gain access to spare embryos from in vitro fertilisation clinics with the consent of donors. Niakan’s team will then begin to examine the role of a set of genes suspected to be in charge of determining the fate of early embryonic cells.

“When the embryo is only about 16 to 32 cells big, each of these cells faces a fork in the road. They can either develop into the foetus or into the protective placenta and egg sac,” explained Sambasivan, who is not involved in Niakan’s research.

This choice is not random but is dependent on whether certain genes are switched on or off in those cells. One gene for example, OCT4, has been shown to be switched on in cells that become foetal cells but switched off in cells that develop into the placenta.

Niakan’s lab will be studying OCT4 and other genes. This could be fundamental to understanding issues like infertility and miscarriages. ““An imbalance created at this point could be the factor that triggers problems like defective placental development resulting in limited success of assisted reproduction techniques,” according to Sambasivan.

Step 6: Understand the difference between editing for research and editing for clinical purposes

Comprehending this difference is the debate’s keystone. It is the latter case that presents the risk because, here, the changes are introduced in an embryo with the intention of letting it continue to develop into a human baby. The changes probably involve correcting a disease-causing gene, but our level of knowledge is still not advanced enough for us to be fully confident that no unintended mutations will take place.

Creating unintended mutant humans may be inevitable if “clinics” claiming to provide gene therapy sprout up without any regulation. Then, of course, there’d be no reason to keep us from skipping over to the realm of the ‘designer baby’, whose genes have been modified to bestow traits that are considered desirable.

In April 2015, a team of Chinese scientists attempted to correct a gene causing beta thalassaemia in human embryos (without governmental approval). They achieved very limited success and concluded that gene editing as we know it isn’t yet ready for the leap into clinical applications. Though the team used only non-viable embryos, which wouldn’t have resulted in a live birth anyway, this episode sent shockwaves across the community and even prompted an international summit to discuss the implications of the CRISPR ‘revolution’.

On the other hand, editing human embryos for research purposes alone, like Kathy Niaken at the Crick Institute wants to, is generally considered okay. Moreover, it can prove crucial to understanding human development better.

The statement that emerged from December’s International Summit on Human Gene Editing had this to say: “If, in the process of research, early human embryos or germline cells undergo gene editing, the modified cells should not be used to establish a pregnancy.” (Emphasis added.)

In their application, Niaken & co. have guaranteed that the embryos will be destroyed in a week and that they won’t be implanting any of the embryos to establish pregnancies. “I’m quite sure that if the project is fully monitored like it will be at the Crick Institute or at other places, where there is a strong ethics committee, it’s possible to avoid misuse,” concluded Sambasivan.