Tag Archives: gene editing

The CRISPR ((clustered regularly interspaced short palindromic repeats)-CAS9 gene-editing technique may cause new genetic damage kerfuffle

Setting the stage

Not unexpectedly, CRISPR-Cas9  or clustered regularly interspaced short palindromic repeats-CRISPR-associated protein 9 can be dangerous as these scientists note in a July 16, 2018 news item on phys.org,

Scientists at the Wellcome Sanger Institute have discovered that CRISPR/Cas9 gene editing can cause greater genetic damage in cells than was previously thought. These results create safety implications for gene therapies using CRISPR/Cas9 in the future as the unexpected damage could lead to dangerous changes in some cells.

Reported today (16 July 2018) in the journal Nature Biotechnology, the study also revealed that standard tests for detecting DNA changes miss finding this genetic damage, and that caution and specific testing will be required for any potential gene therapies.

This CRISPR-Cas9 image reminds me of popcorn,

CRISPR-associated protein Cas9 (white) from Staphylococcus aureus based on Protein Database ID 5AXW. Credit: Thomas Splettstoesser (Wikipedia, CC BY-SA 4.0)[ downloaded from https://phys.org/news/2018-07-genome-crisprcas9-gene-higher-thought.html#jCp]

A July 16, 2018 Wellcome Sanger Institute press release (also on EurekAlert), which originated the news item, offers a little more explanation,

CRISPR/Cas9 is one of the newest genome editing tools. It can alter sections of DNA in cells by cutting at specific points and introducing changes at that location. Already extensively used in scientific research, CRISPR/Cas9 has also been seen as a promising way to create potential genome editing treatments for diseases such as HIV, cancer or sickle cell disease. Such therapeutics could inactivate a disease-causing gene, or correct a genetic mutation. However, any potential treatments would have to prove that they were safe.

Previous research had not shown many unforeseen mutations from CRISPR/Cas9 in the DNA at the genome editing target site. To investigate this further the researchers carried out a full systematic study in both mouse and human cells and discovered that CRISPR/Cas9 frequently caused extensive mutations, but at a greater distance from the target site.

The researchers found many of the cells had large genetic rearrangements such as DNA deletions and insertions. These could lead to important genes being switched on or off, which could have major implications for CRISPR/Cas9 use in therapies. In addition, some of these changes were too far away from the target site to be seen with standard genotyping methods.

Prof Allan Bradley, corresponding author on the study from the Wellcome Sanger Institute, said: “This is the first systematic assessment of unexpected events resulting from CRISPR/Cas9 editing in therapeutically relevant cells, and we found that changes in the DNA have been seriously underestimated before now. It is important that anyone thinking of using this technology for gene therapy proceeds with caution, and looks very carefully to check for possible harmful effects.”

Michael Kosicki, the first author from the Wellcome Sanger Institute, said: “My initial experiment used CRISPR/Cas9 as a tool to study gene activity, however it became clear that something unexpected was happening. Once we realised the extent of the genetic rearrangements we studied it systematically, looking at different genes and different therapeutically relevant cell lines, and showed that the CRISPR/Cas9 effects held true.”

The work has implications for how CRISPR/Cas9 is used therapeutically and is likely to re-spark researchers’ interest in finding alternatives to the standard CRISPR/Cas9 method for gene editing.

Prof Maria Jasin, an independent researcher from Memorial Slone Kettering Cancer Centre, New York, who was not involved in the study said: “This study is the first to assess the repertoire of genomic damage arising at a CRISPR/Cas9 cleavage site. While it is not known if genomic sites in other cell lines will be affected in the same way, this study shows that further research and specific testing is needed before CRISPR/Cas9 is used clinically.”

For anyone who’d like to better understand the terms gene editing and CRISPR-Cas9, the Wellcome Sanger Institute provides these explanatory webpages, What is genome editing? and What is CRISPR-Cas9?

For the more advanced, here’s a link and a citation for the paper,

Repair of double-strand breaks induced by CRISPR–Cas9 leads to large deletions and complex rearrangements by Michael Kosicki, Kärt Tomberg, & Allan Bradley. Nature Biotechnology DOI: https://doi.org/10.1038/nbt.4192 Published 16 July 2018

This paper appears to be open access.

The kerfuffle

It seems this news has affected the CRISPR market. From a July 16, 2018 article by Cale Guthrie Weissman for Fast Company,

… CRISPR could unknowingly delete or alter non-targeted genes, which could lead to myriad unintended consequences. This is especially frightening, since the technology is going to be used in human clinical trials.

Meanwhile, other scientists working with CRISPR are trying to downplay the findings, telling STAT [a life sciences and business journalism website] that there have been no reported adverse effects similar to what the study describes. The news, however, has brought about a market reaction–at least three publicly traded companies that focus on CRISPR-based therapies are in stock nosedive. Crispr Therapeutics is down by over 6%; Editas fell by over 3%; and Intellia Therapeutics dropped by over 5%. [emphasis mine]

Damage control

Gaetan Burgio (geneticist, Australian National University)  in a July 16, 2018 essay on phys.org (originating from The Conversation) suggests some calm (Note: Links have been removed),

But a new study has called into question the precision of the technique [CRISPR gene editing technology].

The hope for gene editing is that it will be able to cure and correct diseases. To date, many successes have been reported, including curing deafness in mice, and in altering cells to cure cancer.

Some 17 clinical trials in human patients are registered [emphasis mine] testing gene editing on leukaemias, brain cancers and sickle cell anaemia (where red blood cells are misshaped, causing them to die). Before implementing CRISPR technology in clinics to treat cancer or congenital disorders, we must address whether the technique is safe and accurate.

There are a few options for getting around this problem. One option is to isolate the cells we wish to edit from the body and reinject only the ones we know have been correctly edited.

For example, lymphocytes (white blood cells) that are crucial to killing cancer cells could be taken out of the body, then modified using CRISPR to heighten their cancer-killing properties. The DNA of these cells could be sequenced in detail, and only the cells accurately and specifically gene-modified would be selected and delivered back into the body to kill the cancer cells.

While this strategy is valid for cells we can isolate from the body, some cells, such as neurons and muscles, cannot be removed from the body. These types of cells might not be suitable for gene editing using Cas9 scissors.

Fortunately, researchers have discovered other forms of CRISPR systems that don’t require the DNA to be cut. Some CRISPR systems only cut the RNA, not the DNA (DNA contains genetic instructions, RNA convey the instructions on how to synthesise proteins).

As RNA [ribonucleic acid] remains in our cells only for a specific period of time before being degraded, this would allow us to control the timing and duration of the CRISPR system delivery and reverse it (so the scissors are only functional for a short period of time).

This was found to be successful for dementia in mice. Similarly, some CRISPR systems simply change the letters of the DNA, rather than cutting them. This was successful for specific mutations causing diseases such as hereditary deafness in mice.

I agree with Burgio’s conclusion (not included here) that we have a lot more to learn and I can’t help wondering why there are 17 registered human clinical trials at this point.

Yes! Art, genetic modifications, gene editing, and xenotransplantation at the Vancouver Biennale (Canada)

Patricia Piccinini’s Curious Imaginings Courtesy: Vancouver Biennale [downloaded from http://dailyhive.com/vancouver/vancouver-biennale-unsual-public-art-2018/]

Up to this point, I’ve been a little jealous of the Art/Sci Salon’s (Toronto, Canada) January 2018 workshops for artists and discussions about CRISPR ((clustered regularly interspaced short palindromic repeats))/Cas9 and its social implications. (See my January 10, 2018 posting for more about the events.) Now, it seems Vancouver may be in line for its ‘own’ discussion about CRISPR and the implications of gene editing. The image you saw (above) represents one of the installations being hosted by the 2018 – 2020 edition of the Vancouver Biennale.

While this posting is mostly about the Biennale and Piccinini’s work, there is a ‘science’ subsection featuring the science of CRISPR and xenotransplantation. Getting back to the Biennale and Piccinini: A major public art event since 1988, the Vancouver Biennale has hosted over 91 outdoor sculptures and new media works by more than 78 participating artists from over 25 countries and from 4 continents.

Quickie description of the 2018 – 2020 Vancouver Biennale

The latest edition of the Vancouver Biennale was featured in a June 6, 2018 news item on the Daily Hive (Vancouver),

The Vancouver Biennale will be bringing new —and unusual— works of public art to the city beginning this June.

The theme for this season’s Vancouver Biennale exhibition is “re-IMAGE-n” and it kicks off on June 20 [2018] in Vanier Park with Saudi artist Ajlan Gharem’s Paradise Has Many Gates.

Gharem’s architectural chain-link sculpture resembles a traditional mosque, the piece is meant to challenge the notions of religious orthodoxy and encourages individuals to image a space free of Islamophobia.

Melbourne artist Patricia Piccinini’s Curious Imaginings is expected to be one of the most talked about installations of the exhibit. Her style of “oddly captivating, somewhat grotesque, human-animal hybrid creature” is meant to be shocking and thought-provoking.

Piccinini’s interactive [emphasis mine] experience will “challenge us to explore the social impacts of emerging biotechnology and our ethical limits in an age where genetic engineering and digital technologies are already pushing the boundaries of humanity.”

Piccinini’s work will be displayed in the 105-year-old Patricia Hotel in Vancouver’s Strathcona neighbourhood. The 90-day ticketed exhibition [emphasis mine] is scheduled to open this September [2018].

Given that this blog is focused on nanotechnology and other emerging technologies such as CRISPR, I’m focusing on Piccinini’s work and its art/science or sci-art status. This image from the GOMA Gallery where Piccinini’s ‘Curious Affection‘ installation is being shown from March 24 – Aug. 5, 2018 in Brisbane, Queensland, Australia may give you some sense of what one of her installations is like,

Courtesy: Queensland Art Gallery | Gallery of Modern Art (QAGOMA)

I spoke with Serena at the Vancouver Biennale office and asked about the ‘interactive’ aspect of Piccinini’s installation. She suggested the term ‘immersive’ as an alternative. In other words, you won’t be playing with the sculptures or pressing buttons and interacting with computer screens or robots. She also noted that the ticket prices have not been set yet and they are currently developing events focused on the issues raised by the installation. She knew that 2018 is the 200th anniversary of the publication of Mary Shelley’s Frankenstein but I’m not sure how the Biennale folks plan (or don’t plan)  to integrate any recognition of the novle’s impact on the discussions about ‘new’ technologies .They expect Piccinini will visit Vancouver. (Note 1: Piccinini’s work can  also be seen in a group exhibition titled: Frankenstein’s Birthday Party at the Hosfselt Gallery in San Francisco (California, US) from June 23 – August 11, 2018.  Note 2: I featured a number of international events commemorating the 200th anniversary of the publication of Mary Shelley’s novel, Frankenstein, in my Feb. 26, 2018 posting. Note 3: The term ‘Frankenfoods’ helped to shape the discussion of genetically modified organisms and food supply on this planet. It was a wildly successful campaign for activists affecting legislation in some areas of research. Scientists have not been as enthusiastic about the effects. My January 15, 2009 posting briefly traces a history of the term.)

The 2018 – 2020 Vancouver Biennale and science

A June 7, 2018 Vancouver Biennale news release provides more detail about the current series of exhibitions,

The Biennale is also committed to presenting artwork at the cutting edge of discussion and in keeping with the STEAM (science, technology, engineering, arts, math[ematics]) approach to integrating the arts and sciences. In August [2018], Colombian/American visual artist Jessica Angel will present her monumental installation Dogethereum Bridge at Hinge Park in Olympic Village. Inspired by blockchain technology, the artwork’s design was created through the integration of scientific algorithms, new developments in technology, and the arts. This installation, which will serve as an immersive space and collaborative hub for artists and technologists, will host a series of activations with blockchain as the inspirational jumping-off point.

In what is expected to become one of North America’s most talked-about exhibitions of the year, Melbourne artist Patricia Piccinini’s Curious Imaginings will see the intersection of art, science, and ethics. For the first time in the Biennale’s fifteen years of creating transformative experiences, and in keeping with the 2018-2020 theme of “re-IMAGE-n,” the Biennale will explore art in unexpected places by exhibiting in unconventional interior spaces.  The hyperrealist “world of oddly captivating, somewhat grotesque, human-animal hybrid creatures” will be the artist’s first exhibit in a non-museum setting, transforming a wing of the 105-year-old Patricia Hotel. Situated in Vancouver’s oldest neighbourbood of Strathcona, Piccinini’s interactive experience will “challenge us to explore the social impacts of emerging bio-technology and our ethical limits in an age where genetic engineering and digital technologies are already pushing the boundaries of humanity.” In this intimate hotel setting located in a neighborhood continually undergoing its own change, Curious Imaginings will empower visitors to personally consider questions posed by the exhibition, including the promises and consequences of genetic research and human interference. …

There are other pieces being presented at the Biennale but my special interest is in the art/sci pieces and, at this point, CRISPR.

Piccinini in more depth

You can find out more about Patricia Piccinini in her biography on the Vancouver Biennale website but I found this Char Larsson April 7, 2018 article for the Independent (UK) more informative (Note: A link has been removed),

Patricia Piccinini’s sculptures are deeply disquieting. Walking through Curious Affection, her new solo exhibition at Brisbane’s Gallery of Modern Art, is akin to entering a science laboratory full of DNA experiments. Made from silicone, fibreglass and even human hair, her sculptures are breathtakingly lifelike, however, we can’t be sure what life they are like. The artist creates an exuberant parallel universe where transgenic experiments flourish and human evolution has given way to genetic engineering and DNA splicing.

Curious Affection is a timely and welcome recognition of Piccinini’s enormous contribution to reaching back to the mid-1990s. Working across a variety of mediums including photography, video and drawing, she is perhaps best known for her hyperreal creations.

As a genre, hyperrealism depends on the skill of the artist to create the illusion of reality. To be truly successful, it must convince the spectator of its realness. Piccinini acknowledges this demand, but with a delightful twist. The excruciating attention to detail deliberately solicits our desire to look, only to generate unease, as her sculptures are imbued with a fascinating otherness. Part human, part animal, the works are uncannily familiar, but also alarmingly “other”.

Inspired by advances in genetically modified pigs to generate replacement organs for humans [also known as xenotransplantation], we are reminded that Piccinini has always been at the forefront of debates concerning the possibilities of science, technology and DNA cloning. She does so, however, with a warm affection and sense of humour, eschewing the hysterical anxiety frequently accompanying these scientific developments.

Beyond the astonishing level of detail achieved by working with silicon and fibreglass, there is an ethics at work here. Piccinini is asking us not to avert our gaze from the other, and in doing so, to develop empathy and understanding through the encounter.

I encourage anyone who’s interested to read Larsson’s entire piece (April 7, 2018 article).

According to her Wikipedia entry, Piccinini works in a variety of media including video, sound, sculpture, and more. She also has her own website.

Gene editing and xenotransplantation

Sarah Zhang’s June 8, 2018 article for The Atlantic provides a peek at the extraordinary degree of interest and competition in the field of gene editing and CRISPR ((clustered regularly interspaced short palindromic repeats))/Cas9 research (Note: A link has been removed),

China Is Genetically Engineering Monkeys With Brain Disorders

Guoping Feng applied to college the first year that Chinese universities reopened after the Cultural Revolution. It was 1977, and more than a decade’s worth of students—5.7 million—sat for the entrance exams. Feng was the only one in his high school to get in. He was assigned—by chance, essentially—to medical school. Like most of his contemporaries with scientific ambitions, he soon set his sights on graduate studies in the United States. “China was really like 30 to 50 years behind,” he says. “There was no way to do cutting-edge research.” So in 1989, he left for Buffalo, New York, where for the first time he saw snow piled several feet high. He completed his Ph.D. in genetics at the State University of New York at Buffalo.

Feng is short and slim, with a monk-like placidity and a quick smile, and he now holds an endowed chair in neuroscience at MIT, where he focuses on the genetics of brain disorders. His 45-person lab is part of the McGovern Institute for Brain Research, which was established in 2000 with the promise of a $350 million donation, the largest ever received by the university. In short, his lab does not lack for much.

Yet Feng now travels to China several times a year, because there, he can pursue research he has not yet been able to carry out in the United States. [emphasis mine] …

Feng had organized a symposium at SIAT [Shenzhen Institutes of Advanced Technology], and he was not the only scientist who traveled all the way from the United States to attend: He invited several colleagues as symposium speakers, including a fellow MIT neuroscientist interested in tree shrews, a tiny mammal related to primates and native to southern China, and Chinese-born neuroscientists who study addiction at the University of Pittsburgh and SUNY Upstate Medical University. Like Feng, they had left China in the ’80s and ’90s, part of a wave of young scientists in search of better opportunities abroad. Also like Feng, they were back in China to pursue a type of cutting-edge research too expensive and too impractical—and maybe too ethically sensitive—in the United States.

Here’s what precipitated Feng’s work in China, (from Zhang’s article; Note: Links have been removed)

At MIT, Feng’s lab worked on genetically engineering a monkey species called marmosets, which are very small and genuinely bizarre-looking. They are cheaper to keep due to their size, but they are a relatively new lab animal, and they can be difficult to train on lab tasks. For this reason, Feng also wanted to study Shank3 on macaques in China. Scientists have been cataloging the social behavior of macaques for decades, making it an obvious model for studies of disorders like autism that have a strong social component. Macaques are also more closely related to humans than marmosets, making their brains a better stand-in for those of humans.

The process of genetically engineering a macaque is not trivial, even with the advanced tools of CRISPR. Researchers begin by dosing female monkeys with the same hormones used in human in vitro fertilization. They then collect and fertilize the eggs, and inject the resulting embryos with CRISPR proteins using a long, thin glass needle. Monkey embryos are far more sensitive than mice embryos, and can be affected by small changes in the pH of the injection or the concentration of CRISPR proteins. Only some of the embryos will have the desired mutation, and only some will survive once implanted in surrogate mothers. It takes dozens of eggs to get to just one live monkey, so making even a few knockout monkeys required the support of a large breeding colony.

The first Shank3 macaque was born in 2015. Four more soon followed, bringing the total to five.

To visit his research animals, Feng now has to fly 8,000 miles across 12 time zones. It would be a lot more convenient to carry out his macaque research in the United States, of course, but so far, he has not been able to.

He originally inquired about making Shank3 macaques at the New England Primate Research Center, one of eight national primate research centers then funded by the National Institutes of Health in partnership with a local institution (Harvard Medical School, in this case). The center was conveniently located in Southborough, Massachusetts, just 20 miles west of the MIT campus. But in 2013, Harvard decided to shutter the center.

The decision came as a shock to the research community, and it was widely interpreted as a sign of waning interest in primate research in the United States. While the national primate centers have been important hubs of research on HIV, Zika, Ebola, and other diseases, they have also come under intense public scrutiny. Animal-rights groups like the Humane Society of the United States have sent investigators to work undercover in the labs, and the media has reported on monkey deaths in grisly detail. Harvard officially made its decision to close for “financial” reasons. But the announcement also came after the high-profile deaths of four monkeys from improper handling between 2010 and 2012. The deaths sparked a backlash; demonstrators showed up at the gates. The university gave itself two years to wind down their primate work, officially closing the center in 2015.

“They screwed themselves,” Michael Halassa, the MIT neuroscientist who spoke at Feng’s symposium, told me in Shenzhen. Wei-Dong Yao, another one of the speakers, chimed in, noting that just two years later CRISPR has created a new wave of interest in primate research. Yao was one of the researchers at Harvard’s primate center before it closed; he now runs a lab at SUNY Upstate Medical University that uses genetically engineered mouse and human stem cells, and he had come to Shenzhen to talk about restarting his addiction research on primates.

Here’s comes the competition (from Zhang’s article; Note: Links have been removed),

While the U.S. government’s biomedical research budget has been largely flat, both national and local governments in China are eager to raise their international scientific profiles, and they are shoveling money into research. A long-rumored, government-sponsored China Brain Project is supposed to give neuroscience research, and primate models in particular, a big funding boost. Chinese scientists may command larger salaries, too: Thanks to funding from the Shenzhen local government, a new principal investigator returning from overseas can get 3 million yuan—almost half a million U.S. dollars—over his or her first five years. China is even finding success in attracting foreign researchers from top U.S. institutions like Yale.

In the past few years, China has seen a miniature explosion of genetic engineering in monkeys. In Kunming, Shanghai, and Guangzhou, scientists have created monkeys engineered to show signs of Parkinson’s, Duchenne muscular dystrophy, autism, and more. And Feng’s group is not even the only one in China to have created Shank3 monkeys. Another group—a collaboration primarily between researchers at Emory University and scientists in China—has done the same.

Chinese scientists’ enthusiasm for CRISPR also extends to studies of humans, which are moving much more quickly, and in some cases under less oversight, than in the West. The first studies to edit human embryos and first clinical trials for cancer therapies using CRISPR have all happened in China. [emphases mine]

Some ethical issues are also covered (from Zhang’s article),

Parents with severely epileptic children had asked him if it would be possible to study the condition in a monkey. Feng told them what he thought would be technically possible. “But I also said, ‘I’m not sure I want to generate a model like this,’” he recalled. Maybe if there were a drug to control the monkeys’ seizures, he said: “I cannot see them seizure all the time.”

But is it ethical, he continued, to let these babies die without doing anything? Is it ethical to generate thousands or millions of mutant mice for studies of brain disorders, even when you know they will not elucidate much about human conditions?

Primates should only be used if other models do not work, says Feng, and only if a clear path forward is identified. The first step in his work, he says, is to use the Shank3 monkeys to identify the changes the mutations cause in the brain. Then, researchers might use that information to find targets for drugs, which could be tested in the same monkeys. He’s talking with the Oregon National Primate Research Center about carrying out similar work in the United States. ….[Note: I have a three-part series about CRISPR and germline editing* in the US, precipitated by research coming out of Oregon, Part 1, which links to the other parts, is here.]

Zhang’s June 8, 2018 article is excellent and I highly recommend reading it.

I touched on the topic of xenotransplanttaion in a commentary on a book about the science  of the television series, Orphan Black in a January 31,2018 posting (Note: A chimera is what you use to incubate a ‘human’ organ for transplantation or, more accurately, xenotransplantation),

On the subject of chimeras, the Canadian Broadcasting Corporation (CBC) featured a January 26, 2017 article about the pig-human chimeras on its website along with a video,

The end

I am very excited to see Piccinini’s work come to Vancouver. There have been a number of wonderful art and art/science installations and discussions here but this is the first one (I believe) to tackle the emerging gene editing technologies and the issues they raise. (It also fits in rather nicely with the 200th anniversary of the publication of Mary Shelley’s Frankenstein which continues to raise issues and stimulate discussion.)

In addition to the ethical issues raised in Zhang’s article, there are some other philosophical questions:

  • what does it mean to be human
  • if we are going to edit genes to create hybrid human/animals, what are they and how do they fit into our current animal/human schema
  • are you still human if you’ve had an organ transplant where the organ was incubated in a pig

There are also going to be legal issues. In addition to any questions about legal status, there are also fights about intellectual property such as the one involving Harvard & MIT’s [Massachusetts Institute of Technology] Broad Institute vs the University of California at Berkeley (March 15, 2017 posting)..

While I’m thrilled about the Piccinini installation, it should be noted the issues raised by other artworks hosted in this version of the Biennale are important. Happily, they have been broached here in Vancouver before and I suspect this will result in more nuanced  ‘conversations’ than are possible when a ‘new’ issue is introduced.

Bravo 2018 – 2020 Vancouver Biennale!

* Germline editing is when your gene editing will affect subsequent generations as opposed to editing out a mutated gene for the lifetime of a single individual.

Art/sci and CRISPR links

This art/science posting may prove of some interest:

The connectedness of living things: an art/sci project in Saskatchewan: evolutionary biology (February 16, 2018)

A selection of my CRISPR posts:

CRISPR and editing the germline in the US (part 1 of 3): In the beginning (August 15, 2017)

NOTE: An introductory CRISPR video describing how CRISPR/Cas9 works was embedded in part1.

Why don’t you CRISPR yourself? (January 25, 2018)

Editing the genome with CRISPR ((clustered regularly interspaced short palindromic repeats)-carrying nanoparticles (January 26, 2018)

Immune to CRISPR? (April 10, 2018)

CRISPR-Cas12a as a new diagnostic tool

Similar to Cas9, Cas12a is has an added feature as noted in this February 15, 2018 news item on ScienceDaily,

Utilizing an unsuspected activity of the CRISPR-Cas12a protein, researchers created a simple diagnostic system called DETECTR to analyze cells, blood, saliva, urine and stool to detect genetic mutations, cancer and antibiotic resistance and also diagnose bacterial and viral infections. The scientists discovered that when Cas12a binds its double-stranded DNA target, it indiscriminately chews up all single-stranded DNA. They then created reporter molecules attached to single-stranded DNA to signal when Cas12a finds its target.

A February 15, 2018 University of California at Berkeley (UC Berkeley) news release by Robert Sanders and which originated the news item, provides more detail and history,

CRISPR-Cas12a, one of the DNA-cutting proteins revolutionizing biology today, has an unexpected side effect that makes it an ideal enzyme for simple, rapid and accurate disease diagnostics.

blood in test tube

(iStock)

Cas12a, discovered in 2015 and originally called Cpf1, is like the well-known Cas9 protein that UC Berkeley’s Jennifer Doudna and colleague Emmanuelle Charpentier turned into a powerful gene-editing tool in 2012.

CRISPR-Cas9 has supercharged biological research in a mere six years, speeding up exploration of the causes of disease and sparking many potential new therapies. Cas12a was a major addition to the gene-cutting toolbox, able to cut double-stranded DNA at places that Cas9 can’t, and, because it leaves ragged edges, perhaps easier to use when inserting a new gene at the DNA cut.

But co-first authors Janice Chen, Enbo Ma and Lucas Harrington in Doudna’s lab discovered that when Cas12a binds and cuts a targeted double-stranded DNA sequence, it unexpectedly unleashes indiscriminate cutting of all single-stranded DNA in a test tube.

Most of the DNA in a cell is in the form of a double-stranded helix, so this is not necessarily a problem for gene-editing applications. But it does allow researchers to use a single-stranded “reporter” molecule with the CRISPR-Cas12a protein, which produces an unambiguous fluorescent signal when Cas12a has found its target.

“We continue to be fascinated by the functions of bacterial CRISPR systems and how mechanistic understanding leads to opportunities for new technologies,” said Doudna, a professor of molecular and cell biology and of chemistry and a Howard Hughes Medical Institute investigator.

DETECTR diagnostics

The new DETECTR system based on CRISPR-Cas12a can analyze cells, blood, saliva, urine and stool to detect genetic mutations, cancer and antibiotic resistance as well as diagnose bacterial and viral infections. Target DNA is amplified by RPA to make it easier for Cas12a to find it and bind, unleashing indiscriminate cutting of single-stranded DNA, including DNA attached to a fluorescent marker (gold star) that tells researchers that Cas12a has found its target.

The UC Berkeley researchers, along with their colleagues at UC San Francisco, will publish their findings Feb. 15 [2018] via the journal Science’s fast-track service, First Release.

The researchers developed a diagnostic system they dubbed the DNA Endonuclease Targeted CRISPR Trans Reporter, or DETECTR, for quick and easy point-of-care detection of even small amounts of DNA in clinical samples. It involves adding all reagents in a single reaction: CRISPR-Cas12a and its RNA targeting sequence (guide RNA), fluorescent reporter molecule and an isothermal amplification system called recombinase polymerase amplification (RPA), which is similar to polymerase chain reaction (PCR). When warmed to body temperature, RPA rapidly multiplies the number of copies of the target DNA, boosting the chances Cas12a will find one of them, bind and unleash single-strand DNA cutting, resulting in a fluorescent readout.

The UC Berkeley researchers tested this strategy using patient samples containing human papilloma virus (HPV), in collaboration with Joel Palefsky’s lab at UC San Francisco. Using DETECTR, they were able to demonstrate accurate detection of the “high-risk” HPV types 16 and 18 in samples infected with many different HPV types.

“This protein works as a robust tool to detect DNA from a variety of sources,” Chen said. “We want to push the limits of the technology, which is potentially applicable in any point-of-care diagnostic situation where there is a DNA component, including cancer and infectious disease.”

The indiscriminate cutting of all single-stranded DNA, which the researchers discovered holds true for all related Cas12 molecules, but not Cas9, may have unwanted effects in genome editing applications, but more research is needed on this topic, Chen said. During the transcription of genes, for example, the cell briefly creates single strands of DNA that could accidentally be cut by Cas12a.

The activity of the Cas12 proteins is similar to that of another family of CRISPR enzymes, Cas13a, which chew up RNA after binding to a target RNA sequence. Various teams, including Doudna’s, are developing diagnostic tests using Cas13a that could, for example, detect the RNA genome of HIV.

infographic about DETECTR system

(Infographic by the Howard Hughes Medical Institute)

These new tools have been repurposed from their original role in microbes where they serve as adaptive immune systems to fend off viral infections. In these bacteria, Cas proteins store records of past infections and use these “memories” to identify harmful DNA during infections. Cas12a, the protein used in this study, then cuts the invading DNA, saving the bacteria from being taken over by the virus.

The chance discovery of Cas12a’s unusual behavior highlights the importance of basic research, Chen said, since it came from a basic curiosity about the mechanism Cas12a uses to cleave double-stranded DNA.

“It’s cool that, by going after the question of the cleavage mechanism of this protein, we uncovered what we think is a very powerful technology useful in an array of applications,” Chen said.

Here’s a link to and a citation for the paper,

CRISPR-Cas12a target binding unleashes indiscriminate single-stranded DNase activity by Janice S. Chen, Enbo Ma, Lucas B. Harrington, Maria Da Costa, Xinran Tian, Joel M. Palefsky, Jennifer A. Doudna. Science 15 Feb 2018: eaar6245 DOI: 10.1126/science.aar6245

This paper is behind a paywall.

Emerging technology and the law

I have three news bits about legal issues that are arising as a consequence of emerging technologies.

Deep neural networks, art, and copyright

Caption: The rise of automated art opens new creative avenues, coupled with new problems for copyright protection. Credit: Provided by: Alexander Mordvintsev, Christopher Olah and Mike Tyka

Presumably this artwork is a demonstration of automated art although they never really do explain how in the news item/news release. An April 26, 2017 news item on ScienceDaily announces research into copyright and the latest in using neural networks to create art,

In 1968, sociologist Jean Baudrillard wrote on automatism that “contained within it is the dream of a dominated world […] that serves an inert and dreamy humanity.”

With the growing popularity of Deep Neural Networks (DNN’s), this dream is fast becoming a reality.

Dr. Jean-Marc Deltorn, researcher at the Centre d’études internationales de la propriété intellectuelle in Strasbourg, argues that we must remain a responsive and responsible force in this process of automation — not inert dominators. As he demonstrates in a recent Frontiers in Digital Humanities paper, the dream of automation demands a careful study of the legal problems linked to copyright.

An April 26, 2017 Frontiers (publishing) news release on EurekAlert, which originated the news item, describes the research in more detail,

For more than half a century, artists have looked to computational processes as a way of expanding their vision. DNN’s are the culmination of this cross-pollination: by learning to identify a complex number of patterns, they can generate new creations.

These systems are made up of complex algorithms modeled on the transmission of signals between neurons in the brain.

DNN creations rely in equal measure on human inputs and the non-human algorithmic networks that process them.

Inputs are fed into the system, which is layered. Each layer provides an opportunity for a more refined knowledge of the inputs (shape, color, lines). Neural networks compare actual outputs to expected ones, and correct the predictive error through repetition and optimization. They train their own pattern recognition, thereby optimizing their learning curve and producing increasingly accurate outputs.

The deeper the layers are, the higher the level of abstraction. The highest layers are able to identify the contents of a given input with reasonable accuracy, after extended periods of training.

Creation thus becomes increasingly automated through what Deltorn calls “the arcane traceries of deep architecture”. The results are sufficiently abstracted from their sources to produce original creations that have been exhibited in galleries, sold at auction and performed at concerts.

The originality of DNN’s is a combined product of technological automation on one hand, human inputs and decisions on the other.

DNN’s are gaining popularity. Various platforms (such as DeepDream) now allow internet users to generate their very own new creations . This popularization of the automation process calls for a comprehensive legal framework that ensures a creator’s economic and moral rights with regards to his work – copyright protection.

Form, originality and attribution are the three requirements for copyright. And while DNN creations satisfy the first of these three, the claim to originality and attribution will depend largely on a given country legislation and on the traceability of the human creator.

Legislation usually sets a low threshold to originality. As DNN creations could in theory be able to create an endless number of riffs on source materials, the uncurbed creation of original works could inflate the existing number of copyright protections.

Additionally, a small number of national copyright laws confers attribution to what UK legislation defines loosely as “the person by whom the arrangements necessary for the creation of the work are undertaken.” In the case of DNN’s, this could mean anybody from the programmer to the user of a DNN interface.

Combined with an overly supple take on originality, this view on attribution would further increase the number of copyrightable works.

The risk, in both cases, is that artists will be less willing to publish their own works, for fear of infringement of DNN copyright protections.

In order to promote creativity – one seminal aim of copyright protection – the issue must be limited to creations that manifest a personal voice “and not just the electric glint of a computational engine,” to quote Deltorn. A delicate act of discernment.

DNN’s promise new avenues of creative expression for artists – with potential caveats. Copyright protection – a “catalyst to creativity” – must be contained. Many of us gently bask in the glow of an increasingly automated form of technology. But if we want to safeguard the ineffable quality that defines much art, it might be a good idea to hone in more closely on the differences between the electric and the creative spark.

This research is and be will part of a broader Frontiers Research Topic collection of articles on Deep Learning and Digital Humanities.

Here’s a link to and a citation for the paper,

Deep Creations: Intellectual Property and the Automata by Jean-Marc Deltorn. Front. Digit. Humanit., 01 February 2017 | https://doi.org/10.3389/fdigh.2017.00003

This paper is open access.

Conference on governance of emerging technologies

I received an April 17, 2017 notice via email about this upcoming conference. Here’s more from the Fifth Annual Conference on Governance of Emerging Technologies: Law, Policy and Ethics webpage,

The Fifth Annual Conference on Governance of Emerging Technologies:

Law, Policy and Ethics held at the new

Beus Center for Law & Society in Phoenix, AZ

May 17-19, 2017!

Call for Abstracts – Now Closed

The conference will consist of plenary and session presentations and discussions on regulatory, governance, legal, policy, social and ethical aspects of emerging technologies, including (but not limited to) nanotechnology, synthetic biology, gene editing, biotechnology, genomics, personalized medicine, human enhancement technologies, telecommunications, information technologies, surveillance technologies, geoengineering, neuroscience, artificial intelligence, and robotics. The conference is premised on the belief that there is much to be learned and shared from and across the governance experience and proposals for these various emerging technologies.

Keynote Speakers:

Gillian HadfieldRichard L. and Antoinette Schamoi Kirtland Professor of Law and Professor of Economics USC [University of Southern California] Gould School of Law

Shobita Parthasarathy, Associate Professor of Public Policy and Women’s Studies, Director, Science, Technology, and Public Policy Program University of Michigan

Stuart Russell, Professor at [University of California] Berkeley, is a computer scientist known for his contributions to artificial intelligence

Craig Shank, Vice President for Corporate Standards Group in Microsoft’s Corporate, External and Legal Affairs (CELA)

Plenary Panels:

Innovation – Responsible and/or Permissionless

Ellen-Marie Forsberg, Senior Researcher/Research Manager at Oslo and Akershus University College of Applied Sciences

Adam Thierer, Senior Research Fellow with the Technology Policy Program at the Mercatus Center at George Mason University

Wendell Wallach, Consultant, ethicist, and scholar at Yale University’s Interdisciplinary Center for Bioethics

 Gene Drives, Trade and International Regulations

Greg Kaebnick, Director, Editorial Department; Editor, Hastings Center Report; Research Scholar, Hastings Center

Jennifer Kuzma, Goodnight-North Carolina GlaxoSmithKline Foundation Distinguished Professor in Social Sciences in the School of Public and International Affairs (SPIA) and co-director of the Genetic Engineering and Society (GES) Center at North Carolina State University

Andrew Maynard, Senior Sustainability Scholar, Julie Ann Wrigley Global Institute of Sustainability Director, Risk Innovation Lab, School for the Future of Innovation in Society Professor, School for the Future of Innovation in Society, Arizona State University

Gary Marchant, Regents’ Professor of Law, Professor of Law Faculty Director and Faculty Fellow, Center for Law, Science & Innovation, Arizona State University

Marc Saner, Inaugural Director of the Institute for Science, Society and Policy, and Associate Professor, University of Ottawa Department of Geography

Big Data

Anupam Chander, Martin Luther King, Jr. Professor of Law and Director, California International Law Center, UC Davis School of Law

Pilar Ossorio, Professor of Law and Bioethics, University of Wisconsin, School of Law and School of Medicine and Public Health; Morgridge Institute for Research, Ethics Scholar-in-Residence

George Poste, Chief Scientist, Complex Adaptive Systems Initiative (CASI) (http://www.casi.asu.edu/), Regents’ Professor and Del E. Webb Chair in Health Innovation, Arizona State University

Emily Shuckburgh, climate scientist and deputy head of the Polar Oceans Team at the British Antarctic Survey, University of Cambridge

 Responsible Development of AI

Spring Berman, Ira A. Fulton Schools of Engineering, Arizona State University

John Havens, The IEEE [Institute of Electrical and Electronics Engineers] Global Initiative for Ethical Considerations in Artificial Intelligence and Autonomous Systems

Subbarao Kambhampati, Senior Sustainability Scientist, Julie Ann Wrigley Global Institute of Sustainability, Professor, School of Computing, Informatics and Decision Systems Engineering, Ira A. Fulton Schools of Engineering, Arizona State University

Wendell Wallach, Consultant, Ethicist, and Scholar at Yale University’s Interdisciplinary Center for Bioethics

Existential and Catastrophic Ricks [sic]

Tony Barrett, Co-Founder and Director of Research of the Global Catastrophic Risk Institute

Haydn Belfield,  Academic Project Administrator, Centre for the Study of Existential Risk at the University of Cambridge

Margaret E. Kosal Associate Director, Sam Nunn School of International Affairs, Georgia Institute of Technology

Catherine Rhodes,  Academic Project Manager, Centre for the Study of Existential Risk at CSER, University of Cambridge

These were the panels that are of interest to me; there are others on the homepage.

Here’s some information from the Conference registration webpage,

Early Bird Registration – $50 off until May 1! Enter discount code: earlybirdGETs50

New: Group Discount – Register 2+ attendees together and receive an additional 20% off for all group members!

Click Here to Register!

Conference registration fees are as follows:

  • General (non-CLE) Registration: $150.00
  • CLE Registration: $350.00
  • *Current Student / ASU Law Alumni Registration: $50.00
  • ^Cybsersecurity sessions only (May 19): $100 CLE / $50 General / Free for students (registration info coming soon)

There you have it.

Neuro-techno future laws

I’m pretty sure this isn’t the first exploration of potential legal issues arising from research into neuroscience although it’s the first one I’ve stumbled across. From an April 25, 2017 news item on phys.org,

New human rights laws to prepare for advances in neurotechnology that put the ‘freedom of the mind’ at risk have been proposed today in the open access journal Life Sciences, Society and Policy.

The authors of the study suggest four new human rights laws could emerge in the near future to protect against exploitation and loss of privacy. The four laws are: the right to cognitive liberty, the right to mental privacy, the right to mental integrity and the right to psychological continuity.

An April 25, 2017 Biomed Central news release on EurekAlert, which originated the news item, describes the work in more detail,

Marcello Ienca, lead author and PhD student at the Institute for Biomedical Ethics at the University of Basel, said: “The mind is considered to be the last refuge of personal freedom and self-determination, but advances in neural engineering, brain imaging and neurotechnology put the freedom of the mind at risk. Our proposed laws would give people the right to refuse coercive and invasive neurotechnology, protect the privacy of data collected by neurotechnology, and protect the physical and psychological aspects of the mind from damage by the misuse of neurotechnology.”

Advances in neurotechnology, such as sophisticated brain imaging and the development of brain-computer interfaces, have led to these technologies moving away from a clinical setting and into the consumer domain. While these advances may be beneficial for individuals and society, there is a risk that the technology could be misused and create unprecedented threats to personal freedom.

Professor Roberto Andorno, co-author of the research, explained: “Brain imaging technology has already reached a point where there is discussion over its legitimacy in criminal court, for example as a tool for assessing criminal responsibility or even the risk of reoffending. Consumer companies are using brain imaging for ‘neuromarketing’, to understand consumer behaviour and elicit desired responses from customers. There are also tools such as ‘brain decoders’ which can turn brain imaging data into images, text or sound. All of these could pose a threat to personal freedom which we sought to address with the development of four new human rights laws.”

The authors explain that as neurotechnology improves and becomes commonplace, there is a risk that the technology could be hacked, allowing a third-party to ‘eavesdrop’ on someone’s mind. In the future, a brain-computer interface used to control consumer technology could put the user at risk of physical and psychological damage caused by a third-party attack on the technology. There are also ethical and legal concerns over the protection of data generated by these devices that need to be considered.

International human rights laws make no specific mention to neuroscience, although advances in biomedicine have become intertwined with laws, such as those concerning human genetic data. Similar to the historical trajectory of the genetic revolution, the authors state that the on-going neurorevolution will force a reconceptualization of human rights laws and even the creation of new ones.

Marcello Ienca added: “Science-fiction can teach us a lot about the potential threat of technology. Neurotechnology featured in famous stories has in some cases already become a reality, while others are inching ever closer, or exist as military and commercial prototypes. We need to be prepared to deal with the impact these technologies will have on our personal freedom.”

Here’s a link to and a citation for the paper,

Towards new human rights in the age of neuroscience and neurotechnology by Marcello Ienca and Roberto Andorno. Life Sciences, Society and Policy201713:5 DOI: 10.1186/s40504-017-0050-1 Published: 26 April 2017

©  The Author(s). 2017

This paper is open access.

Managing risks in a world of converging technology (the fourth industrial revolution)

Finally there’s an answer to the question: What (!!!) is the fourth industrial revolution? (I took a guess [wrongish] in my Nov. 20, 2015 post about a special presentation at the 2016 World Economic Forum’s IdeasLab.)

Andrew Maynard in a Dec. 3, 2015 think piece (also called a ‘thesis’) for Nature Nanotechnology answers the question,

… an approach that focuses on combining technologies such as additive manufacturing, automation, digital services and the Internet of Things, and … is part of a growing movement towards exploiting the convergence between emerging technologies. This technological convergence is increasingly being referred to as the ‘fourth industrial revolution’, and like its predecessors, it promises to transform the ways we live and the environments we live in. (While there is no universal agreement on what constitutes an ‘industrial revolution’, proponents of the fourth industrial revolution suggest that the first involved harnessing steam power to mechanize production; the second, the use of electricity in mass production; and the third, the use of electronics and information technology to automate production.)

In anticipation of the the 2016 World Economic Forum (WEF), which has the fourth industrial revolution as its theme, Andrew  explains how he sees the situation we are sliding into (from Andrew Maynard’s think piece),

As more people get closer to gaining access to increasingly powerful converging technologies, a complex risk landscape is emerging that lies dangerously far beyond the ken of current regulations and governance frameworks. As a result, we are in danger of creating a global ‘wild west’ of technology innovation, where our good intentions may be among the first casualties.

There are many other examples where converging technologies are increasing the gap between what we can do and our understanding of how to do it responsibly. The convergence between robotics, nanotechnology and cognitive augmentation, for instance, and that between artificial intelligence, gene editing and maker communities both push us into uncertain territory. Yet despite the vulnerabilities inherent with fast-evolving technological capabilities that are tightly coupled, complex and poorly regulated, we lack even the beginnings of national or international conceptual frameworks to think about responsible decision-making and responsive governance.

He also lists some recommendations,

Fostering effective multi-stakeholder dialogues.

Encouraging actionable empathy.

Providing educational opportunities for current and future stakeholders.

Developing next-generation foresight capabilities.

Transforming approaches to risk.

Investing in public–private partnerships.

Andrew concludes with this,

… The good news is that, in fields such as nanotechnology and synthetic biology, we have already begun to develop the skills to do this — albeit in a small way. We now need to learn how to scale up our efforts, so that our convergence in working together to build a better future mirrors the convergence of the technologies that will help achieve this.

It’s always a pleasure to read Andrew’s work as it’s thoughtful. I was surprised (since Andrew is a physicist by training) and happy to see the recommendation for “actionable empathy.”

Although, I don’t always agree with him on this occasion I don’t have any particular disagreements but I think that including a recommendation or two to cover the certainty we will get something wrong and have to work quickly to right things would be a good idea.  I’m thinking primarily of governments which are notoriously slow to respond with legislation for new developments and equally slow to change that legislation when the situation changes.

The technological environment Andrew is describing is dynamic, that is fast-moving and changing at a pace we have yet to properly conceptualize. Governments will need to change so they can respond in an agile fashion. My suggestion is:

Develop policy task forces that can be convened in hours and given the authority to respond to an immediate situation with oversight after the fact

Getting back to Andrew Maynard, you can find his think piece in its entirety via this link and citation,

Navigating the fourth industrial revolution by Andrew D. Maynard. Nature Nanotechnology 10, 1005–1006 (2015) doi:10.1038/nnano.2015.286 Published online 03 December 2015

This paper is behind a paywall.