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Since the end of the 20th century, CRISPR has been the most discussed topic in the scientific community. There are consistent predictions that this gene-editing technology has the potential to transform our planet. Compared to older and other popular tools used for genetic engineering, CRISPR-Cas9 is cheap, easy to use, precise, and remarkably powerful. The applicability of CRISPR is also wide-ranging from human biology, agriculture, and microbiology.

Researchers are still trying to figure out how CRISPR can be used to change the world for the better. However, the power to alter DNA brings along many ethical questions and concerns. In this article, we will be looking at the latest CRISPR Research Trends and tips on how to build up a successful career in CRISPR.

1. Use of CRISPR To Treat Genetic Errors That Cause Disease

One of the rare, genetic, heart diseases is Hypertrophic cardiomyopathy (HCM), affecting roughly 1 in every 500 people worldwide. This occurs because of mutations in a number of dominant genes, which leads the heart tissues to stiffen, and gives rise to chest pain, weakness, and, in severe cases, sudden cardiac arrest. Still, there is a hope, that in the due course of time, we will be able to use gene editing to cure this disease once and for all.

Attempts had been made in 2017 when the Oregon Health and Science University researchers used CRISPR to delete one of these defective genes in a number of viable human embryos. They injected 54 embryos with the CRISPR-Cas9 machinery 18 hours after fertilization. The results indicated that 36 of the embryos did not show any chance of developing the disease. Other 13 were having 50 percent chances of inheriting HCM. On further reducing the chances of inheritance, it was found that only one was a mosaic.

2. Algorithm-based approach to improve efficacy

As per the report published in November 2018 in the leading journal Nature Biotechnology, CRISPR-Cas9 will enable scientists to predict the best sequences as a target to make gene editing more reliable, cheaper, less time consuming and more efficient. In this attempt, the researchers analyzed over 40,000 pairs of different target DNA and guide RNA deeply, and carried out CRISPR-Cas9 gene editing in different cells. It was analyzed that the repair depended on the exact sequence of DNA and guide and discovered that it was reproducible within the same sequence. Thereafter, a machine learning computational tool, called FORECasT was generated, that enabled them to predict the repaired sequence, using the targeted DNA sequence alone.

In a similar attempt, a machine-learning algorithm in Delphi has been created that predicts how human and mouse cells respond to CRISPR-induced breaks in DNA. It was discovered that cells themselves repair broken genes in precise and inevitable manners, typically even returning mutated genes back to their healthy version. In fact, the researchers were able to successfully correct mutations in cells taken from patients with one of two rare genetic disorders.

3. Elimination of the Pathogens via CRISPR Cas9

Even after 38 years of discovery of HIV, scientists still haven’t found a cure. CRISPR can turn out to be a hopeful measure in this regard. In 2017, an experiment was carried out on mice to increase resistance to HIV. A team of Chinese researchers successfully implemented this by replicating a mutation of a gene that effectively prevents the virus from entering cells. This mutation has been found to occur naturally in a small percentage of people. Artificially it can be induced using CRISPR, which will considerably bolster HIV resistance in humans in the future. In another approach, scientists from North Carolina used CRISPR to engineer bacteriophages, to develop a proven, safe method for treating harmful bacteria.

4. Recreation of species using CRISPR

Harvard geneticist George Church and his team have invested their valuable 11 years to recreate the DNA blueprint of the extinct mammoth. DNA from mammoths preserved in Arctic permafrost has been used to discover that only 44 genes are separating mammoths from elephants.

It is an attempt to save endangered elephants they are trying to create hardier mammoth hybrids that is much more cold-resistant. The researchers also look forward to inserting non-mammoth genes, which will prevent them from growing tusks, to prevent poaching, as well as new DNA to allow them to eat a wider diet.

5. CRISPR Could Create New, Better & Healthier Foods

Mushrooms that do not turn brown have already been created with the help of CRISPR technology. Attempts have been made by the scientists from Cold Spring Harbor Laboratory in New York to edit the genes which determine fruit size, branching architecture and, the shape of the plant for a greater harvest in tomato.

Before that, we should know the difference between GM crops and gene-edited crops. While traditional GMOs are made by inserting foreign DNA sequences into a crop’s genome, makes precise alterations to genes in specific locations of the native genome.

The leading company, DuPont Pioneer is trying to bring its “waxy” gene-edited corn into the U.S. market by 2020. The U.S. Department of Agriculture regulation has already approved gene-edited mushroom, since it doesn’t contain genomes from viruses or bacteria, declaring it the first CRISPR-edited organism to be green-lit.

6. Possibility to Eradicate Mosquitoes

Discoveries have given access to tackle the sleep menace ‘mosquito’ in the most astonishing ways. Scientists at the University of California, Riverside have disrupted target genes in multiple locations of the mosquito’s genes, using gene drive technology, by impairing the mosquito’s flight and vision. Success was achieved in postponing mosquito development, shortening the animal’s lifespan, retarding egg development, and diminishing fat accumulation. Other researchers at the Imperial College London, have tried another unique way to get rid of a female mosquito, by interfering with how they reproduce.

Current approaches in arthropods are based on delivering the gene-editing Cas9 on to eggs by embryonic microinjection, a tough and inefficient method that works in a mere a tiny low range of species. As per Jason Rasgon, faculty member of zoological science and unwellness medicine, Penn State school of Agricultural Sciences, microinjection will harm the eggs, and it needs costly instrumentality and coaching to implement.

To address these limitations, the researchers developed ReMOT Control—Receptor-Mediated Ovary Transduction of Cargo. This is the technique that will deliver a Cas9 system to the target by a simple injection into the blood of feminine arthropods, wherever it is often introduced into the developing eggs via receptors within the ovary. Rasgon explained that in ovary and egg maturation, mosquitoes and alternative arthropods synthesize food proteins, that are secreted into the blood and brought up into the ovaries. The assumption is to bypass the requirement for embryonic microinjection to attain ordering gene editing within the embryo.

7. Designer babies

The first edit in the genes of a human embryo in vitro was made by China in 2015 using CRISPR + IVF, which sparked global outcry and pleas not to make a baby using the technology. As per the reports, a team at the Southern University of Science and Technology lead by He Jiankui had been recruiting couples so as to create the designer babies. They had planned to eliminate a gene called CCR5 hoping to make the offspring resistant to HIV, smallpox, and cholera. They have successfully brought two healthy, little Chinese girls named Lulu and Nana into the globe in November 2018. He Jiankui said that his team performed “gene surgery” on embryos created from their parents’ sperm cell and eggs to shield the youngsters from the human immunological disorder virus, HIV, that causes AIDS. Their father is HIV-positive.

Indian Research Community is all set to adapt for this trending gene editing technology of CRISPR Cas9. In fact, a lot of research is already being conducted at Govt research labs and private biopharma companies with respect to CRISPR Research Trends – to tackle one re more issues faced globally related to health, agriculture and Food technology.

List of Research Institutes in India Where CRISPR research is being conducted:

• IARI – Indian Agricultural Research Institute (IARI)
• NIPGR – National Institute of Plant Genome Research
• JNU – Jawaharlal Nehru University
• DU – Delhi University
• NBRI – National Botanical Research Institute
• NABI – National Agri-Food Biotechnology Institute
• JUIT – Jaypee University of Information Technology
• ILS – Institute of Life Sciences, BBSR
• NII – National Institute of Immunology
• NBRC – National Brain Research Centre
• IIT – Indian Institute of technology – All Locations
• IIsc – Indian Institute of Science, Bengaluru
• NRCPB – National Research Centre on Plant Biotechnology
• CSIR-NEIST – CSIR North East Institute of Science and Technology
• CSIR-IICB – CSIR Indian Institute of Chemical Biology
• CSIR NCL – CSIR-National Chemical Laboratory
• TIFR – Tata Institute of Fundamental Research
• NCBS – National Center For Biological Sciences
• CCMB – Centre for Cellular & Molecular Biology

List of Biopharma companies in India Where CRISPR research is being conducted:

• Reliance LifeScience
• Thermo Fisher
• Bayer
• Syngene
• Merck
• GE Healthcare

A lot of job opportunities in this field exists for which you need to apply for vacancies in CRISPR related research projects at either research institutes or Biotech companies. But all of these jobs require a minimum qualification of being a postgraduate with NET / GATE qualification if you are joining a research lab or preferably a PhD degree (given priority).

• So, the first step here is to grab a PhD position for yourself after clearing any of the exams that can help you get admission as well as guarantee you a fellowship. E.g. CSIR-JRF, ICMR-JRF, etc.
• You need to select for a potential guide, who himself is well aware of this new technique and can provide you with necessary facilities and guidance.
• Next, one should develop a research problem which has novel applications, that includes the use of CRISPR and which has not been standardized properly. But always keep in mind that you require something to look upon, when in doubt. So, try to select a research problem, which may have been successfully implemented in some other way in some related model organism.
• Now, you should also have a basic understanding of genetic engineering and molecular biology protocols as well. E.g. like cloning, PCR, qPCR, RNA extraction, cDNA synthesis, immunochemistry techniques like western blotting, immunofluorescence. If there is no possibility to keep all these techniques in your research work, you can always opt for hands-on training courses or workshops which can be availed from any of the reputed laboratories.
• It may require additional hand-on-expertise on mammalian cell culture- experience handling multiple cell lines, transfection, electroporation, transduction, generation of stable cell lines, single cell clonal selection, RNAi mediated gene knockdown, transient and stable expression of proteins in mammalian cells.
• Again, in order to get experience in related techniques or CRISPR itself, you can join as a Junior Research Fellow in reputed laboratories.
• Once you have completed your M.Sc. / Ph.D. research work successfully, you can apply for a patent as well, along with the publications in reputed journals. These publications will give you an upper hand while applying for the jobs.
• Moreover, you can apply for Postdoctoral positions in similar fields, as limited knowledge may give you perfection in a particular field, but it will not give you exposure. The more you learn, the better are the chances for you to get a salary hike or promotion or possibly a government job as a scientist.

Let me remind you that CRISPR will offer you highly paid jobs (Research associate Salary US $ 40000). Since his field is still in its infancy, you will get ample opportunities to prove yourself.

Recent developments have proven CRISPR to extremely versatile, precise and safe. However, the technology is still in its adolescence. Till date, we are facing technological and ethical hurdles that are not allowing us to feed the hunger-stricken planet, remove genetic disorders, or conserve extinct animal species back to life. But we are making progress in a satisfactory manner. But one thing is clear- CRISPR is a technology which in near future will be adapted globally looking to its diverse applications but of course, the practice should be done under strict guidelines to avoid its misuse.

So are you CRISPR job ready yet?

Millions of people suffer from devastating genetic disorders like cancer, sickle cell anemia, cystic fibrosis, muscular dystrophy, Huntington’s disease and many more. Can you imagine the pain and suffering that could be avoided if these diseases could simply be cured by rewriting the genetic code of patients? That is how promising the CRISPR-Cas9 gene-editing technology is!

One of the hottest developments in recent years has been the CRISPR gene editing process. CRISPR stands for “clustered regularly interspaced short palindromic repeats” and it’s a way to delete or insert certain genes, or even chemically repair DNA. Its possible applications are arguably endless and there’s a lot of current research happening in chemistry departments all over the world. But there’s one factor some might be missing, and that is the legal side of CRISPR.

The US Patent Office is currently conducting hearings on a CRISPR Patent War – a dispute between the University of California and the Broad Institute at Harvard and MIT. Two biochemists at UC Berkeley filed a patent for the process in 2012 and published their work showcasing their process. Feng Zhang from the Broad Institute was also working on CRISPR and filed a patent after the Berkeley team but he paid to have the US Patent Office expedite his review (which is legal) and thus was issued the first CRISPR patent. The debate has become the definition of CRISPR. The Patent Office has said it’s specific to eukaryotes, cells in plants animals and humans, which Zhang’s work involved. The Berkeley team’s work involved prokaryotes, which are only in bacteria and do not have the potential applications that eukaryotes do. The debate between the two uses of CRISPR could shape the future of technology. A patent dispute of this magnitude is likely to affect research globally.

CRISPR in a very short period of time became one of the most exciting scientific breakthroughs in today’s world. If we look back at the early days of CRISPR discovery, we come to know, that neither Francisco Mihika nor Philipp Horvath had the setup to undertake groundbreaking research. They were merely going about doing routine tasks as part of their jobs or investigating something that just caught the eye. The truth is breakthroughs are most of the times the summation of countless small discoveries. Even you might be working on any small but crucial discovery right now; the manuscript that you write for journals may be picked up someone somewhere in the scientific community to eventually build upon it.

Multiple discoveries about CRISPR were made by many scientists throughout the years and the reason why CRISPR can grow so fast today is that each discovery was communicated to everyone effectively through journals and publications. The scientific community shares one another’s knowledge, successes and failures and science advances as a whole. While Collective progression is an excellent way to accelerate scientific development, it does cause a unique problem – in the discovery of CRISPR it is exactly because so many scientists participated in CRISPR’s development with each having made significant contributions it is impossible to pinpoint a particular person and declare he or she discovered CRISPR. That is the problem that has been unfolding in recent years as scientists scramble to claim patent rights to CRISPR.

Many scientists consider their work to be the significant milestone in CRISPR’s discovery but so many important discoveries were made based on previous discoveries which were based on even earlier discoveries. Instead of a single scientist discovery of CRISPR, the situation is more of a group of scientists in the discovery of CRISPR. This makes it extremely difficult to conclude who should have the patent rights for CRISPR! Since the technology has so much potential, whoever owns the rights to the process will likely have a billion dollar asset in their pocket.

The CRISPR Patent History

• The CRISPR mechanism was first published in the year December 1987 by Amemura, Ishino, Makino, Nakata, Shinagawa, Takase, Wachi at Osaka University.
• On 18 Jan 2000, more clustered repeats of DNA were identified in other bacteria and archaea, termed as Short Regularly Spaced Repeats (SRSR) by Mojica, Diez-Villasenor, Soria, Juez at the University of Alicante and University Miguel Hernandez. The term CRISPR-Cas9 was published for the first time by Mojica, Jansen, Embden, Gaastra, Schouls at Utrecht University in the year March 2002.
• Later Jennifer Doudna and Jillian Banfield started investigating CRISPR at the University of California Berkeley Regulators debate about what restrictions should be enforced with CRISPR/Cas9, the technology that has become the subject of a major patent dispute.
• Dupont had filed the first application to patent the technology in March 2007 (WO/2007/025097). The patent application covers the use of technology to develop phage-resistant bacterial strains for food production, feeds, personal care products, cosmetics, and veterinary products. Since then, patents have been filed by 3 heavily financed Biotechnology start-up companies and half a dozen universities.

UC Berkeley vs MIT – CRISPR Patent War

In the US, 2 major competing patent claims have been filed. One on 25 May 2015, grounded in the work led by Jennifer Doudna at the University of California, Berkeley, and Emmanuelle Charpentier, originally at the University of Vienna and presently at the Helmholtz Centre for Infectious Research in Germany. The application has 155 claims and covers numerous applications for a variety of cell types (US Patent Application No. PCT/US2013/032589).

The second was filed for the work of Feng Zhang by MIT-Harvard Broad Institute on 12 December 2012 which focused on the use of CRISPR/Cas9 for genome editing in eukaryotic cells. A fast-track status was given and the patent was granted on 15 April 2014 (US Patent No. 8,697,359). Charpentier and the UC and Vienna filed a challenge to the patent with the US Patent and Trademark Office, in April 2015. Four years after it entered into a legal battle with the Broad Institute due to a crossover between patents filed by the two parties, the University of California (UC) will soon gain its third patent on the gene-editing technology known as CRISPR.

The US Patent and Trademark Office at Alexandria Virginia, Vienna has had hearings about the CRISPR Cas 9 patent interference, with attorneys representing the University of California on one side and the Broad Institute of MIT and Harvard on the other side just give their case for why they should hold the CRISPR patent. This is a genome editing technology that people have said will change the world. CRISPR potentially can engineer super crops, snip out genetic diseases from humans and even make designer animal models for research. Billions of dollars are at stake now, according to experts. The litigious timeline starts way back in May 2012 when a group of biochemist and molecular biologists including Jennifer Doudna and Emmanuelle Charpentier filed their first CRISPR patent.

A month later, they published their work in a scientific journal, thus officially kicking off the CRISPR craze. While Doudna and Charpentier are quickly rising to CRISPR fame so was Feng Zhang from the Broad Institute. Zhang filed his first CRISPR patent in December 2012, and then quickly published a paper famously demonstrating the first use of CRISPR and mouse in human cells. In a perfectly legal move, the Broad Institute paid to expedite the review of their patent and in April 2014, Zhang was the first to receive an approved patent for CRISPR mediated cell editing, even though he submitted his application after the Berkeley group. Berkeley countered by petitioning for a patent interference, which is a legal proceeding where federal patent court judges determine who made the invention first. The Patent Office did approve Berkeley’s petition but defining CRISPR as a system specifically used in eukaryotes; which are organisms whose chromosomes are found inside a nucleus; such as plants, animals and humans; Not to forget what experts think; that eukaryotes are where the major buck lies at.

Doudna and Charpentier’s patent had CRISPR technology used only in prokaryotes, such as bacteria. Attorneys argued that the judges would have messed over the understanding of the word eukaryote. They said that if CRISPR has worked for prokaryotes, it would obviously work in eukaryotes too. The UC patent even cited the potential for using CRISPR and cells dozens of times. Berkeley’s attorney in their exact words has said that “no special sauce required” to move CRISPR into eukaryotes. Doudna’s own words were turned against her, in a defend by the Broad Institute. She had told the press that she expected many frustrations moving CRISPR from prokaryotes to eukaryotes and that she was unsure if it would even work. Zhang’s patent specifically showed how to use CRISPR in eukaryotic cells. There’s a spectrum of possible outcomes from this hearing the simplest is that one side wins the other side loses.

CRISPR Patent War Possible Outcomes

The losing team is likely to appeal though and an appeal would take the case out of the hands of patent judges with molecular biology backgrounds and send it to federal circuit judges that are unlikely to have any science training at all which may be an unappealing prospect for CRISPR stakeholders. But if Broad institute gets to win this, Berkeley could still get its patent approved, just that it would only cover the prokaryotic uses of CRISPR which look way less lucrative.

Another possible outcome is a sort of tie. The judges declared that both sides filed patents for the same invention and this would kick off a second phase of the case where both sides will have to submit lab notebooks and testimonies to figure out who actually had first thought of the CRISPR technology. There is also a possibility that CRISPR or other gene editing tools will improve so much by the time that judges decide their decision will not really matter.

So, what lies ahead? That depends partly on the appetite of those bankrolling the fight. Neither the UCB nor the Broad Institute is directly paying for this battle. It is being funded by biotech companies instead, which have taken out licenses to the key intellectual property assets in each organization. They have already spent tens of millions of dollars and they are not even in the most expensive phase yet. The costs could easily shoot up to hundreds of millions if the two organizations start to sue each other on grounds of infringement of patent claims. If the companies involved aren’t certain about how large those fees will ultimately be, and who they will need to pay licensing fees to, this may stand in the way of a much-needed innovation. This ongoing uncertainty is a big problem for commercial applications of gene-editing.

Impact of CRISPR Patent War On Us!

Why should all these CRISPR Patent War matters to a layperson? Of course, it should! We all eat food medicine and get treatments which are products of Biotechnology. Considering that CRISPR’s immense potential for real-world applications such as possibly rectifying mutations that cause cancer and hence curing cancer without chemotherapy. So obviously, whoever manages to claim rights to CRISPR; will be able to control the direction of its development deployment and access to the public. Who knows maybe one day we will depend on medicine or treatments enabled by CRISPR.

So whether you will eventually pay $13.50 a pill for an essential drug or 750 dollars a pill will depend on who is granted the patent. Take, for example, the case of price increase of 5000 percent for Daraprim in 2015, a drug patented and FDA approved in 1953 and it becomes all too easy to see how grants on CRISPR in the 2020s now will affect you and your family’s medical treatments in the 2040s. This is what makes the result of this patent dispute so highly anticipated.

We are still in the developing days of CRISPR technology and there is a lot more to understand and work both in and out of the lab. With the patents, licensing CRISPR may become more straightforward or at least clearer and more organized.

The current atmosphere surrounding CRISPR IP rights may be having a lot of litigation and contention around the core patents. But, one thing is certain, that the pace of innovation to create new nucleases, new ways to edit and more predictability means that the future of these technologies certainly looks promising. The future still remains a little way off—the dust is still settling down.

It all started when a Chinese researcher claimed that he had used the gene-editing technique CRISPR–Cas9 to modify human embryos in order to make them HIV resistant and after implanting them into a woman, have made the first CRISPR-edited babies, twin girls named Lulu and Nana. Those two became the first human beings born with genetic modifications that were directed by human beings. It provoked an international outcry; it is like a bombshell revelation.

And it just gets worse by the day. It seems that there were a lot of technical errors and ethical blunders in this alleged creation of the world’s first gene-edited infants.

You may have heard about him in my other podcast on CRISPR’s potential uses, where I had mentioned him to be using the CRISPR/Cas9 system to modify the genes of twin girls, so that they are born with a deletion in the CCR5 gene, thus conferring resistance to HIV infection.

He Jiankui ! He became the centre of a global firestorm on November 25th, 2018, when he made this fact known to the world.

Although he has been sharply criticized for flouting international recommendations and going against ethics rules, it is notable that he did not keep his gene-editing ambitions entirely secret.  Infact, the young researcher had widely telegraphed his plans and shared aspects of his plans among senior American scientists and ethicists, hoping to win their endorsement, and seeking advice, and help with publications. Dr. He had explained the experiment behind it at an international gene-editing summit in Hong Kong. In fact, he also revealed that yet another early pregnancy is also under way.

Chinese scientist He Jiankui speaks at a human genome editing summit in Hong Kong on Nov. 28, 2018. He announced an experiment on twins that raised a range of ethical questions.

Although it is still unclear, if his claims are correct, as there is no definitive evidence, if he was actually successful in modifying the girl’s genes, yet he was flooded with a swift series of strong reactions and negative comments. CSIRSPR pioneer Jennifer Doudna, was actually “horrified” on hearing this, and according to NIH Director Francis Collins, this experiment was “profoundly disturbing”.  Even Julian Savulescu, an ethicist, who feels that gene-editing research is a “moral responsibility”, has described Dr, He’s work to be “monstrous”.

Dr. Kiran Musunuru from University of Pennsylvania, an expert who has no role in the experiment considers it unethical. He feels that we still have a lot of work to do to proof and establish that the procedure is actually safe. He said that no babies should be born at this point of time following the use of this technology, and that this is too early and too immature. He and many mainstream scientists say that this type of gene editing should not be attempted yet because it could make permanent damages to DNA that could affect future generations or be harmful if other genes are affected.

It is extremely important to balance the potential benefits with the potential risks for the people involved. In cases where the potential risks are substantially higher than the benefits, which are in the case of Dr. He’s incident, is not ethical at all.

He’s choice to alter the particular CCR5 gene has also been criticized by many scientists, partly because of the other ways available to stop people from contracting HIV, like using caesarean sections to deliver the babies of mothers with the virus. Critics feel that other diseases, like Huntington’s disease of Tay-Sachs disease would be more obvious targets for elimination through editing embryonic genomes.

Dr. He was aiming at mimicking a mutation existing in about 10% of Europeans, which helps to protect them from HIV infection. But it may be possible that He might have caused mutations in other parts of the genome, which could lead to unpredictable health consequences. Also, CCR5 is believed to be helping people fight off the effects of various other infections, such as West Nile virus. So, if the gene is disabled, the twins could actually become vulnerable, although it may years to identify those health effects. But if they do suffer, and a link is found with He’s experiment, then He could be sentenced to imprisonment on charges of practising medicine illegally, says Zhang Peng, a criminal-law scholar at Beijing Wuzi University.

Gene editing has been attempted to treat serious diseases in people, but those changes are only in that person. It is not allowed in the US and many other countries on embryos intended for pregnancy.

Shoukhrat Mitalipov, a reproductive biologist at the Oregon Health & Science University in Portland, fears that this controversy may affect factors such as funding and regulatory approvals. Mitalipov works on repairing mutated genes in human embryos, and hopes to edit out heritable diseases once day with this approach. The US government prohibits federal funding for such experiments, still Mitalipov and a few other US researchers have somehow managed to get other grant money for their work. This incident makes Mitalipov cautious, like whether this will cause them to face a backlash

He’s experiment leaves an array of other unanswered questions, like whether the prospective parents were properly informed of the risks; why He chose CCR5 modification when clearly there were other, proven methods for HIV prevention; why he chose to do the experiment with couples in which the men have HIV, as it’s a known fact that women with HIV have a higher chance of passing the virus on to their children; and what were the risks of knocking out CCR5, a gene which is normally present in people, and which could have some  necessary but still unknown functions. All these points actually outweighed the benefits in this case.

Chinese authorities’ investigation found that He Jiankui had broken national regulations in his controversial work on gene-edited babies. He was even fired by his university. The decision was announced by the Southern University of Science and Technology in Shenzhen, in China’s Guandong province on 21st January 2019. This was followed by an investigation by the provincial health authorities, the Guagdong health ministry, which were tasked by the national health ministry to examine the He affair. They had found that He broke the national regulations against usage of gene-editing for reproductive purposes. They even found that He’s experiment went against to the. During the recruitment of participants, Dr. He had used forged ethics-review documents and had swapped blood samples to skirt laws and national regulations which forbids people with HIV from using assisted reproductive technologies. This allegation was reported for the first time in the Xinhua article. He is fully blamed for this, and is criticized for doing all these in pursuit of “fame and fortune”, deliberately evading oversight.

The Xinhua article states that He’s gene-editing activities were “clearly prohibited by the state”, although it doesn’t mention any punishable crimes and exactly which specific laws or regulations have been broken. The article also reports that He and others implicated in the case are going to be dealt with seriously and that the police might now get involved.

The article even confirms many details of the case for the first time, like starting from June 2016.  According to it, Dr. He had put together a team that, from March 2017, recruited eight couples, which consisted of an HIV-positive father and an HIV-negative mother. After which He’s team had edited the genes of embryos from at least two couples. In also reported that one other woman involved in the experiment is currently pregnant with a gene-edited embryo.

Keeping in mind the conclusions by Guangdong team, China’s education ministry have announced that they are going to collaborate with other ministries and departments in building a better regulatory system for overseeing science that will help in improving the ethical review of scientific projects. Stricter laws could be around the corner.

Officials at Stanford University are also enquiring some of the several high-profile scientists whom He had consulted, they want to understand what liabilities or risks Stanford may have in connection with the controversial medical experiment.

Few scientists, including the CRISPR pioneer Feng Zhang and stem-cell biologist Paul Knoepfler, have also called for a temporary moratorium on such embryonic gene editing experiments, in the wake of He’s bombshell.

What Dr. He is actually criticized for, is not for his attempt of germ-line editing, but because he had neglected adequate safety testing and did not follow standard procedures in procuring participants. He hadn’t even responded to Nature’s multiple attempts to contact him.

So what’s next for Human Gene-Editing? Dr. He have taken a giant leap into an era in which science could rewrite the gene pool of future generations, by engineering mutations into human embryos and by altering the human germ line. He has also flouted established norms for safety and human protections along the way. The experiments have attracted so much attention, that it could easily alter research in the coming years. He’s actions could practically stall the responsible development of gene editing babies.

There is also a concern about how the future of this field can be affected by the public scrutiny, i.e whether or not researchers will look at altering the germ-line anymore. Negative focus is of course not good. Yet some predict that this incident may in fact propel human gene editing forwards. According to Jonathan Kimmelman, who specializes in human trials of gene therapies at McGill University in Montreal, the definitive actions of this scandal can expedite global cooperation on the science and its oversight, which would definitely stimulate meaningful advances in this area.

On the other hand if you see, there is actually nothing wrong in helping human beings!This is evolution happening right before our eyes! The Future of the human race is here! Scientists are the celebrities of the future! To be clear, no researcher or scientist ever say that CRISPR is bad for baby for sure. It is more of a question of whether there’s unknown side effect. But someone has to get this started, the sooner the better, so may be He’s attempt should be appreciate that he took the risk and got it done.

Many brand new technologies come with small unknown risk, but we still have to look at the overall pro and cons. Even now there are people who still argue that electromagnetic wave is very bad for health. Imagine if those people had the upper hand back then, and banned the release of radio technology. There would be no radio, GPS, wifi, cellular communications, etc.

But in the end, it’s the society who will decide how to move forward. The more people can afford to have their genes changed; more funds can go to the project. The more receptive society is towards the programme, the more flexible it is for the rich to alter their genes. It’s a win win scenario for both rich and poor, There should also be a government subsidy helping the poor family to have their babies’ gene changed!

So what do you feel about this? Is it even justified to be called as a scandal after all? Or you completely support the negative reviews raised on Dr. He’s work.

Making scientific headlines for the past few years, all we can say about CRISPR technology is that, it is no less than a medical revolution bestowed upon mankind. It is by far the most efficient and accurate method to edit a cell’s genome.

CRISPR technology uses very specific DNA scissors, which can be an invaluable tool for correcting genetic mutations that cause deadly diseases like HIV, cystic fibrosis, and cancer, just to name a few. CRISPR can absolutely work wonders. On one hand, it opens up a myriad of new avenues for gene engineering, and on other, CRISPR/Cas9 gene editing method could bring frightening ethical challenges into healthcare in the future.

CRISPR, an integral part of the bacteria defence system, are made up of short palindromic DNA sequences that are repeated along the CRISPR molecule and are regularly-spaced.

Present between these sequences are “spacers”, which are foreign DNA sequences from organisms that have previously attacked the bacteria.

Bacteria produce enzymes to fight off the constant assault from viruses. Whenever an invading virus is killed by the bacterial enzymes, the viral DNA is processed into little pieces, which is then stored in those CRISPR spaces between the repeats, thus serving as genetic memory.

It is like a “molecular most-wanted gallery” having records of all the enemies the microbe has encountered through its short life.

You may wonder why it is present only in bacteria. It is because of the oldest war on Earth – bacteria versus viruses.

The cleverest part of this technology is that the bacteria use the genetic information stored in these CRISPR spaces to fend off future attacks. In case there is a new infection, the CRISPR sequence undergoes transcription, including spacers and Cas genes, creating a single-stranded RNA, called as CRISPR RNA, which contains copies of the invading viral DNA sequence in its spacers.

When these CRISPR RNAs come across a virus, they see if the virus’s genome matches to that of the stored information. And if there is a match, the Cas9 enzyme starts cleaving the virus’s DNA to destroy the targeted viral material and thus neutralize the threat.

The trick is to make use of this CRISPR-Cas9 systems’ recognition of specific DNA sequences and apply it several purposes. Instead of viral DNA as spacers, scientists use their specific gene of interest. So, if any gene’s sequence known, it can act just like a spacer for the system and guide the Cas9 protein to a DNA matching sequence.

Crispr-Cas9 makes it easy, cheap, and fast to move genes around, literally any genes, in any living organism, from bacteria to humans.

So what can CRISPR be used for?

So many things!!

According to food scientist Rodolphe Barrangou, CRISPR-Cas 9 is like a word processing software. It is like correcting genetic typos, you can remove or replace a wrong gene; just how you would correct a misspelled word, or even add a whole sentence or cut out a whole paragraph.

CRISPR-cas 9 system allows researchers to perform gene knock-out, gene insertions or knock-ins, DNA free gene editing (using only RNA or protein components, and transient gene silencing

One of Crispr’s biggest advantages is that it can work on every living entity. That kind of power makes Jennifer Doudna, the world-famous scientist behind CRISPR, feel like she has opened Pandora’s box.

At the most basic level, CRISPR can knock out individual genes and see which traits are affected accordingly, making it much easier for researchers to figure out what different genes in different organisms actually. Although we have had a complete map of the human genome since 2003, we don’t really know all those gene’s functions yet. CRISPR can aid in speeding up the genome screening, and genetics research could advance massively as a result.

The most interesting fact is that CRISPR is actually a pretty broad term. Scientists have discovered that there are numerous CRISPRs, not just one. So when people talk about CRISPR, they usually refer to the CRISPR/Cas9 system. In recent years, researchers like Feng Zhang have found other types of CRISPR proteins which also work as gene editors. Cas13, for example, can edit RNA.

The real fun and, potentially, the real risks can arise using CRISPRs to edit various plants and animals. That is what a recent paper in Nature Biotechnology by Barrangou and Doudna talks about, in which a flurry of potential future applications of CRISPRs is listed:

AGRICULTURE

Let’s talk about CRISPR uses in editing crops to be tastier, more nutritious, or even better survivors of heat and stress.

Since an early time, it was realized that this technology can be put to use in crops to improve traits, such as yield, plant architecture, plant aesthetics, and disease tolerance.

Wang and team from Syngenta Biotechnology, China have been editing rice genome by designing several CRISPR sgRNAs and have successfully deleted fragments of the dense and erect panicle gene in an Indica rice line.

Yupeng Cai , leading a team of researchers in the Chinese Academy of Agricultural Sciences also used the CRISPR-Cas9 system to induce mutations on an integrator in the photoperiod flowering pathway of soybean. The modified soybean plants showed late flowering, resulting in increased vegetative size. Moreover, the mutation was also found to be stably inherited in the following generation.

A research team led by Shouwei Tian in Beijing Key Laboratory of Vegetable Germplasm Improvement, used CRISPR-Cas9 to target the phytoene desaturase in watermelon, to achieve the albino phenotype.

Through CRISPR-Cas9, researchers from the Chinese Academy of Agricultural Sciences and National Centre for Citrus Variety Improvement and Southwest University have developed citrus plants resistant to a serious disease of citrus, citrus canker caused by Xanthomonas citri. Here, CRISPR is used to target the promoter of the gene which promotes canker development in citrus.

Cold Spring Harbor Laboratory, in collaboration with various other research institutions, achieved mutations in tomato’s flowering suppressor gene to manipulate photoperiod response, resulting in a rapid flowering and enhanced growth habit of field tomatoes, thus having a quick burst of flower production and early yield.

CRISPR has been successfully used to solve a range of food-related concerns for both consumers and growers such as reduced-gluten wheat that could be tolerated by those with sensitivities, a mushroom that doesn’t brown when bruised or cut, soybeans lower in unhealthy fats, and even protecting the global chocolate supply—candymaker Mars is trying to bolster cacao’s ability in fighting off a virus causing devastation of the crop in West Africa.

Korean scientists are working to see if CRISPR could help bananas survive a deadly fungal disease.

How many of you love peanut butter, but cannot have it, as you are allergic to them? Well, CRISPR can also potentially be used to snip out the allergens in peanuts.

Gene editing is relatively easy for those with proper training and basic lab facilities and not tightly controlled by a few companies, hence it might allow developing nations to grow drought-free corn or nutrient-enriched vegetables without buying expensive seeds from large multinational firms. It saves time for growers trying to  methodically cross generations of plant species to eventually get the desired trait—Crispr helps to cut off years from that process.

Some researchers have shown that CRISPR can even create hornless dairy cows which are a huge advance for animal welfare.

Major companies like Monsanto and DuPont have recently begun licensing CRISPR technology, hoping to develop valuable new crop varieties. This technique may not entirely replace traditional GMO techniques, which aim at transplanting genes from one organism to another. But CRISPR can definitely allow scientists to identify genes for certain traits and also to insert desired traits into crops more precisely than traditional breeding, which is a much messier way of swapping in genes.

However successfully breeding new varieties could still take years of testing; according to Pamela Ronald, a plant geneticist at the University of California Davis.

USE IN MEDICINE

Researchers are now using CRISPR/Cas9 to edit the human genome and trying to knock out genetic diseases like hypertrophic cardiomyopathy, as shown in the new Nature paper. Using it on mutations that cause Huntington’s disease or cystic fibrosis are also looked at, as well as treating breast and ovarian cancers by targeting  it on the BRCA-1 and 2 mutations are also talked about.

However, these have only been tested on cells in the lab. It will be a while to overcome few hurdles before anyone starts clinical trials on actual humans.

One of the hurdles is that the Cas9 enzymes can occasionally “misfire” and edit DNA in unexpected places, which might lead to cancer or even create new diseases in human cells. CRISPR’s ability to wreak havoc on DNA has been “seriously underestimated”; as righty said by geneticist Allan Bradley of England’s Wellcome Sanger Institute,

Although there have been major advances in improving CRISPR precision and reducing these off-target effects, still scientists are urging caution on human testing. There is still plenty of challenging work left to be done on the delivery of the editing molecules to particular cells.

Scientists around the world are already using this rapidly emerging technique as gene-editing tools in several of their projects. Several global research and development companies, for example, Columbia University Medical Center (CUMC) and University of Iowa scientists have used CRISPR to repair a genetic mutation causing retinitis pigmentosa (RP), which is an inherited condition causing the retina to degrade and leading to blindness in at least 1.5 million cases worldwide.

CRISPR’s use in gene-editing is just the tip of the iceberg. CRISPR can be used as a tool to turn genes on and off. Stanley Qi, working at Stanford University found a way to “mess up” the working of the DNA scissors, actually blunting them, and thus creating a “dead” version of Cas9 that can’t cut anything at all.

The CRISPR-Cas9 system has been adapted to generate technologies called CRISPRi (CRISPR interference) and CRISPRa (CRISPR activation), or to tune their activity over a 1,000-fold range. These utilize nuclease-deactivated Cas9 (dCas9) that instead of generating a DSB, targets the genomic regions resulting in RNA-directed transcriptional control. CRISPRi utilizes dCas9 that complexes with gRNA to target promoter regions for transcriptional repression, or knockdown, of the gene.

CRISPRa on the other hand uses dCas9 fused to different transcriptional activation domains, to direct them to promoter regions by special gRNAs that recruit additional transcriptional activation domains to upregulate expression of the target gene.

So now instead of a precise set of scissors, which cuts a specific gene, we also have a precise delivery system, which can also CONTROL a specific gene. So basically it not only acts as an editor but can also act as a controller of a tiny entity from outside. Genius, and scary as well, isn’t it?

There also is an option to use Cas9 for live / in-vivo fluorescent imaging of cells, specifically chromosome dynamics.

Several such Cas9 modifications can be impactful like Cas9 can be made less toxic in stem cells; HDR can be increased with comparison to NHEJ and specificity can be increased to the point that single nucleotide differences can be discriminated.

Recently, there was a breakthrough in this technology with the use of a CRISPR that is capable of breaking RNA. This RNA version of CRISPR, which is based on a certain enzyme known as C2c2 was developed by researchers at the Massachusetts Institute of Technology (MIT).

Researchers could influence gene activity as well as the production of protein in the body, by manipulating the RNA. This not only allows them to effectively turn the process up or down, it also allows them to switch it on or off without affecting the genetic codes stored in the RNA. Now better forms of treatment can be developed for targeting specific malignancies in the body, such as Huntington’s disease.

A newer CRISPR tool, named SHERLOCK, can detect tiny amounts of viruses. It is a new diagnostic system that can detect attomolar levels of viruses in a sample and also distinguish Zika from its close relative, dengue. This exquisitely sensitive and specific tool exploits a variation of CRISPR, and promises to help detect diseases that other diagnostics miss, and it’s simple and cheap to use.

About 3.2 billion people – nearly half of the world’s population – are at risk of malaria, according to The World Health Organisation (WHO) estimates. Hence it is overtly important to fight and prevent the disease, by fighting off its primary transmitter: the infected mosquitoes.

Therefore, scientists have used CSIRPR to create mosquitoes that are almost entirely resistant to the parasite that causes malaria. A segment of mosquito DNA was removed, and during the repairing of the genome, it was tricked into getting replaced with a specially engineered DNA construct.

CRISPR is the ultimate weapon against cancer, which occurs when cells refuse to die and keep multiplying in various places in our bodies, while hiding from our immune system. With CRISPR, our immune cells can be edited to improve them against cancer cells and to help them kill these malevolent entities in time. In the future, Getting rid of cancer could mean just an injection in the future. Amazing isn’t it!

Recently, there was a miraculous incident. As a last resort to save a girl suffering from lymphoblastic leukemia, doctors had decided to use gene-editing technologies, where they altered the T-cells of a donor so that they can effectively locate and kill leukemia cells without attacking the patient’s organism. They had actually used another method, TALEN, but in any case, it turned out to be a huge success.

Duchenne ’s muscular dystrophy is caused by a mutation which prevents the body to produce a critical protein in the development of muscle tissue, the dystrophin protein. Patients with this syndrome lose the ability to walk at an early age, and often die from respiratory complications or heart failure.

Thankfully, this syndrome is retraceable to one specific mutation of a gene, thus scientists are experimenting with the use of CRISPR in finding an effective treatment and has been successful in treating mice models. Definitely looks quite promising

CRISPR has been used in University of California for studies in gene therapy. They were able to correct mutations associated with the genetic disease, β-thalassemia by creating induced pluripotent stem cells (iPSCs) from the β-thalassemia patients. The team corrected mutations in the human hemoglobin beta (HBB) in patient iPSCs, resulting in gene-corrected iPSCs with restored expression of the HBB gene, which can be used for gene therapy.  CRISPR also has potential to make algae produce sustainable energy.

From curing genetic diseases and perhaps even remove vulnerability to other illnesses like HIV it could even make cancers unable to attack the cells they would normally affect.

As we all know, that bacteria evolve continuously and gains resistance to antibiotics. There is a desperate requirement for effective antibiotics, but, it’s difficult and costly to develop fresh antibiotics for deadly infections. But CRISPR/Cas9 systems may be used to eradicate certain bacteria more precisely than ever. While others researchers are working on CRISPR systems to target viruses such as HIV and herpes.

An astounding use of CRISPR is in an unnerving concept called as gene drives, which are used to modify not just a single organism but an entire species.

So this is how it works:  When any organism mates, there’s a 50-50 chance that it will pass on a particular gene to its offspring. Using CRISPR, these odds can be altered so that there’s a nearly 100 percent chance that a particular gene gets passed on. Thus it will ensure that an altered gene propagates throughout an entire population.

Through this technique, mosquitoes can be genetically modified to produce only male offsprings and then use gene drive to push that trait through an entire population. Over time, this could wipe out an entire population of malaria-spreading mosquitoes.

What Could CRISPR Do Tomorrow? Where could CRISPR get us in the future

Boosting Human Intelligence? Editing humans?

With eradication of diseases, are we close to designing perfectly healthy humans? Or may be with identifying and cutting out an aging gene, are we going to have eternal youth and live for 2-3 hundred years?

It seems that CRISPR could mean the ultimate solution to cure HIV and ultimately AIDS, as scientists have used it to cut HIV cells out of living cells of patients.

CRISPR/CAS9 could also develop new-age drugs, meaning a revolution for the pharmaceutical industry. A $300 million joint venture has been recently announced by the company Bayer AG and start-up CRISPR Therapeutics for developing CRISPR-based drugs to treat heart disease, blood disorders, and blindness.

Who knows, it may be possible to treat cancer or AIDS through a pill or an injection, in a couple of decades.

You may also expect arrival of super plants, like jasmines blossoming the whole year or harvesting seasonal fruits all year

In the era of smart phones, smart homes, smart cars and artificial intelligence, it seems to be a smart idea to try to boost the intelligence of actual people as well.

We might also one day use CRISPR to create Designer-babies to may be select for athleticism or superior intelligence. It’s not entirely farfetched.

However, researchers aren’t even close to being able to do this. According to Doudna, the reality is we don’t understand enough yet about the human genome, how genes interact, which genes give rise to certain traits. She however believes that it will change over time.

Another intriguing question is can CRISPR be used to grow tails in humans. 

It would be hard, but not totally unimaginable. Just that we are unaware of which genes are actually involved in the development of a tail, if the disappearance of the tail is required for normal development and if the presence of a tail would affect other aspects of development.

CRISPR has become such a fast-moving field that Jennifer Doudna herself finds it difficult to keep up with the updates.

CRISPR is also helping Geneticist George Church and his team to put mammoth DNA into elephant cells, with the end goal of creating some sort of mammoth-like elephant, bringing back a version of an extinct species.

Scientists at Columbia University Medical Centre have created the world’s smallest tape recorder out of E.coli. The state-of-the-art recording device is from modified pieces of DNA called plasmids. It can not only record their interactions with the environment but time-stamp the events

Very recently, a researcher in China, He Jankui, used the CRISPR/Cas9 system to modify the genes of twin girls, so that they are born with a deletion in the CCR5 gene, thus confering resistance to HIV infection. Those two became the first human beings born with genetic modifications that were directed by human beings.

Humans have started to direct their own evolution!

In her book named “A crack in creation”, Jennifer Doudna, talks about CRISPR’s monumental discovery and describes its power to reshape the future of all life and warns of its use. Thanks to this startling and unprecedented discovery, the dreams of genetic manipulation have become a stark reality now.

The hardest part is moving from the theoretical potential to real, practical applications, moving them into a clinical setting.

Though it is a Herculean task, there’s a lot of motivation both on the part of academic scientists as well as companies in figuring out the steps to make it possible.

Three public companies; Crispr Therapeutics AG, Editas Medicine Inc. and Intellia Therapeutics Inc. have seen their shares skyrocket over the past year even before an actual clinical trial. It is even forecasted by Citi analysts, that the global market for Crispr technologies may reach $10 billion by 2025.

CRISPR technology slides almost frictionlessly into our modern culture attracting swarms of investors to bring genetically engineered creations to market.

As a relatively young technique, it has already received a lot of attention in recent years due to its range of applications, including biological research, breeding and development of agricultural crops and animals, and human health applications.

Wheat is rendered invulnerable to killer fungi like powdery mildew, hinting at using engineered staple crops.  Crispr has been used to alter the DNA of yeast so that it consumes plant matter and excretes ethanol, promising an end to reliance on petrochemicals. Several start-ups  devoted to Crispr have been launched. Crispr R&D has spun up in international pharmaceutical and agricultural companies. Crispr makes you see a either a gleaming world of the future,or a Nobel medallion, or may be dollar signs, depending on what kind of a person you are.

Yes, CRISPR is revolutionary, and like all revolutions, it’s perilous.

It could make the long thought idea of designer babies, invasive mutants, species-specific bioweapons, and a dozen other apocalyptic sci-fi tropes come true, allowing genetics researchers to conjure everything anyone has ever worried they would. CRISPR brings with it all new rules for the actual practice of research in life sciences.

Thousands of scientists are working on Crispr. But all of them will not be as cautious.  And as Doudna’s postdoc, Martin Jinek quotes: You can’t stop science from progressing, Science is what it is. I totally agree:  Science gives people power. And power IS unpredictable.