TLDR: This is thoroughly informative book at the right era when Genetic engineering, CRISPR is at the forefront of COVID fight. Also celebrating two extraordinary women nobel laureates and shows intricacies including political wrangling between top universities such as Broad vs Harvard vs Berkeley. Fascinating read.
Rating 9/10 .
Summary
Jennifer Doudna in 2020 when she picked her son Andy from the cancelled Robotics contest at Berkley due to outbreak of Covid decided something. She will use CRISPR the gene editing tool that she pioneered alongside other famous bio scientists worldwide to find a cure to the COVID menace . In 2018 her chinese counterpart did a groudbreaking innovation with CRISPR synthesizing lab designed twin babies who were removed of the HIV causing gene from the embryo been passed typically from mother. CRISPR is a natural phenomenon in bacteria over several millenia a defense mechanism they use to fight viruses. Author notes how 'Wonderful nature truly is' which is a theme of this book and CRISPR research. This though raised ethical conundrum , COVID was a perfect storm to push the CRISPR boundaries further. Doudna as with other notable scientists had a very difficult childhood , her father who had a PHd himself moved the kids to a small Hawaii town and Douda as an introverted blonde felt out of place and was constantly derided by the local kids. Doudna in 3rd grade thankfully moved to a new forgiving township and environment and a smaller class of only 20 from the 60 earlier . This proved the 'Big fish in a small pond' hypothesis adequately . Doudna first excelled in social studies related to american history and later built confidence into her natural habitats of Maths, sciences . On advise of teachers she jumped a grade skipping 5th in the process due to her precocity. Doudna wanted and craved for a stronger ally and in Lisa Henley she found one. Lisa showed her how to look directly at bullies and counter them head on . Lisa was also an outdoorsy gutsy girl into hiking , trekking , skiing and beach . She proudly announced to her class her ambition to be a world class sky diver and this boosted Doudna's confidence further and she learnt to challenge status quo and look beyond the regular. One example is playing half back in soccer team that needed stamina and which most girls in her age did not want to play. Jennifer's dad noted her extraordinary capacity to absorb things and decided to harness her unlimited potential through books. He introduced several good library books from his lists that specifically focused on Female achievers and scientists like Madame curie and the 1950 Gene revolutionary findings where Denise Barlow played a significant part . This inspired Doudna further in her scientific quest.
The book now moves to some lessons / vignettes from history , focusing on how Charles Darwin through his 'Galapagos' missions at age 22 made groundbreaking discoveries . Darwin found species of 'Finches' that certainly evolved and mutated from 'Fruit seeking' to 'Nut / worm ' seeking and what he noticed as 'Natural selection' or Sports as he termed. He felt his theories of evolutions might be deemed heretic and hesitated to publish them and only when another scientist Alfred Russel Wallace in the same era had similar observations. Both Darwin and Wallace decided to exchange notes and publish findings in the very same conference. Gregor mendel solidified Darwins findings further by coming up with the concept of 'Gene' through his Austrian farm exploits. Mendel found 'Dominant' traits in flowers overpowering 'Recessive' traits in a series of interesting experiments and Gene answered Darwin's natural selection and evolution perfectly. After computers, Internet / cloud / mobile , 2020 and beyond is seen as the century of Biology and human genome thanks to the myriad possibilities of virology , CRISPR etc.
Watson Kirik at cambridge worked on DnA under grant from Sir. Hugh lawrence. After involvement of Franklin a lady Crystallized Xray expert from Kings disproving much of the initial thesis of Watson, Kirik .. Lawrence lost patience and asked them to move away from DnA research . Franklin as watson put it was atypical in an era where sexism was at its peak and women were actively looked down for Scientific contributions. Franklin wore asexual , neutral clothes and Watson consistently believed she was attractive if only she took interest in clothes and make up ! Watson referred to her as Rosy and Franklin who had enough to deal already actively refused to collaborate with Watson despite her Xray niche expertise . Linus Pauling who had then won a nobel for discovering Protein anatomy wrote a paper on DnA confirming its helical structure but much else on the paper reeked of patch work . Watson Kirik latched on to this paper confirming their helical suspicion but wanted Franklin's xrays to confirm . Watson barged into Kings campus and in a way stole some critical xrays from franklin thanks to his involvment with another Phd researcher named Wilkins who wanted a cut on the project. Watson kirik and Wikins published a game changing 950 page paper on DnA confirming its double helical structure and properties that confirmed it was the key for copying genes in organisms. This finding was game changing for biology probably next best thing after Darwin's theory of evolution . In the 50s alongside turing's maths and computer revolution thanks to bits , DnAs ACTG sub components helped enter a field of 'Codes' and computerization of literally every information . Biology suddenly was buzzing and Kirik, watson, wilkins won a nobel in 1958 . The noble committee avoided embarassment as they can only award 3 ppl at a time and since Franklin died a year earlier to ovarian cancer at a young age of 37 - everyone forgot about her involvement conveniently. Franklin's achievements though lived in memories of many erudite young girls most notably Doudner herself in present day.
Doudner got inspired by the amazing story of Kirik, Watson and Franklin . The story had everything power , politics, rivalry, innovation and groundbreaking research - If there were any doubts on doudners mind the book made her believe women can be scientists. On her own journey she got encouragement from her French teacher to pursue chemistry after her hyper masculine conservative japanese high school teacher asked her not to do Chemistry exams . Luckily sanity prevailed and doudner did ace her high school chemistry exams and got a physics-chemistry undergrad program at caltech. She ended up in one of the most quiry labs of Don who never told students the 'Process' and allowed them to arrive at their own learnings. This right place at the right time got doudner absorbed with 'Discovering' things, one such experiments of bacteria in a pencillin petri dish led to a world class paper and accolade for doudner. This in no small measures paved the way for a reluctant PG admission into Harvard. At Harvard doudner got the shock of her life seeing how the chemistry faculty were full of young latino , spanish speakers breaking the myth of old white men. Doudner was allowed space to challenge the norm and that is how she radically cloned a bacteria with her 'own approach' . Watson by now started the famous 'Human genome' project in the prestigious long island cold spring harbor labs / university. Cold spring harbor did not exactly have a full checkered past due to their involvement in 'Eugenics' research in the early 20th century. Watson himself had a personal agenda with 'Human genome research' i.e mapping all the 3 billion plus Dna strand in the human body since his son Rufus though intelligent was a 'Schrizophenic' child who tried to commit suicide in WTC towers. Watson wanted to understand how 'genes' contribute to mental health through this. Doudner joined the Human genome project in the 90s and so start another exciting phase for Biology.
The human genome project got great attention from Bill clinton who thought they were close to understanding 'God's design'. Craig venter the celera founder whose company had made groundbreaking innovations around DNA sequencing was now a celebrity due to his media savvy persona. Francis collins meanwhile inherited the human genome project from Watson in the cold harbor labs and took it few notches up , however venter & collins each had parts of puzzle and dint want to collaborate , it took clinton and white house to bring them together for 'Public good of humanity' . They eventually uncorked and claimed the project could read all of 3billion DNA strands and the coded instructions it contained - this ability to read was seen as a marvel and collins, venter won the Nobel. Venter reluctantly got collins to share the Times cover with him. Meanwhile the ability to write or modify the DnA codes is seen as the ultimate holy grail and not much traction happened on it via DnA research . It turned out the clue was with RnA as a 1988 nobel discovery confirmed some RnA (Ribo nuclei acid) strands could replicate themselves and become enzymes which were coined as Ribozymes. Doudner was fascinated about RnA since these were the messengers that read the codes (DnA) in a cell's nuclei and helped form proteins which actually was the catalyst for every human life action . Proteins formed tissues, skin, bone, muscles etc. Proteins helped transact chemical reactions into biomechanical actions and the key catalyst being enzymes.
Doudner divorced Griffith in her pursuit of RnA research as both though scientists had very different demeanours and purposes. Doudner wanted scientific breakthrough as the sole motive of her life but Griffith was a 9-5 science guy and broad swathe of interests including books, music etc. Doudner was becoming a big name in RnA research circle for her post graduate work and publications at Harvard under the notable Jack Szostak and they devoted time and energy to read RnA and find out how it functions, replicates and more importantly can we chemically synthesize it. Jack Szostak famously believed in not following a research which had 1000 ppl in it and that appealed to Doudner's philosophy. Suddenly due to their work and insights RnA became focus in the 90s. There was however a question of how the first RnA came into being - most prevalent theory was that earth produced the right set of chemicals to tune into the first RnA that was the fulcrum for DnA and life itself. Jack and Doudner published several papers on RnA mutations and potential synthesizes of Ribozymes . However Doudner was crippled by what she saw was lack of visuals in the research with Jack and so she found an alternate collaborator at Boulder colorado. Tomas Cech was a reputed researcher on RnA and expert in Xray crystallography. Cech needed Doudna's ideas and Doudna wanted cech's visuals and both together made several breakthroughs on the RnA crystal landscape. Doundna had a trusted student named Jamie cate who moved with her across to Yale and they jointly synthesized and preserved RnA crystals (Ribozymes) through Yales Cryogenic techniques. Cate got romantically involved with Doudna post her divorce with Griffith and he was exactly the same type as her , could go hours together researching or running experiments as opposed to Griffith's 9-5 dedication. Cate married Doudna in 2000 at Hawaii. Ribozymes could now be synthesized in lab and understood lot better. Doudna keeps a letter from James watson sent before her Phd in 1987 framed to this day. Doudna feels Watson though has a bad reputation worldwide for his 'Eugenics' blabbering , she holds him in high esteem for mentoring and taking special attention in women scientists. Doudna post 1996 when she and cate revealed how RNA can be crystallized and synthesized in labs spent considerable time with her ailing dad - Doudna says she is who she is because of her dad's unflinching belief in her and nudge for her persistence to science. Doudna's dad kept the scientific curiousity alive till his last breath and showed remarkable interest and grasp in Doudna's work. Doudna , Lisa (Her best friend), first biology teacher in Hawaii all took Martin Doudna's ashes and scattered at helo a small town in hawaii in a river. Lisa mentions there was an endemic hawk circling the ashes , which depicted gods according to Native american myth.
Doudna was seduced by several universities notably yale which offered multiple incentives and roles to keep both Doudna and cate in , also harvard where doudna started off offered her a plum position as head researcher in the newly started chemistry and chemical biology dept. Doudna settled for Berkely in California and decided not to go to Boston as Cate was pursuing research at MIT , for conflict of interest reasons. Doudna started her research in Berkely on the RnA side first focusing on the 1998 groundbreaking revelations by two biologists that won them nobel . The 1998 paper stated how petunia's DNA genes could not be modified despite injecting necessary MrNA enzymes. The phenomenon that led to 'RnA interference' i.e MRNA being sabatoged or the 'Gene part' being silenced could spark revolution in Gene editing and curing gene related ailments. Doudna started studying the 'Dicer' part of the new RNA that was interfering with the MrNA and applied crystal biology on it. However there was something far more revolutionary happening in Gene editing unknown to doudna and mainstream biologists. Francis Mojica who Phd in US and several universities returned to his native spain to study Bacteria in lab . In his pursuit of E coli in the spanish beaches and studying their 'Patterned RNA' , Mojica stumbled onto something significant and earth shaking. E coli used these patterns and a 'Gene associate' as immunity against viruses with similar code . Some bacteria even showed capability of altering the 'codes' according to their invaders. This patterned repeats was called by Mojica as CRISPR (Clustered Regulatory Interspersed Short Palindromic repeats ) after corresponding with a Dutch biologist and vetting the name with his spanish wife (who said it was fun like how a pup would be called) . Mojica honestly only pursued it since he marvelled nature and was curious how bacteria countered virus for millions even billions of years . He struggled to publish his findings which were not taken seriously by leading editors including nature and ended up publishing in an obscure molecular biology paper after much prodding . However his paper was found by other notable researchers including Jill bamfield a genial Australian who contacted Doudna promptly and others prompting dozens of CRISPR researches worldwide.
In 2006, geomicrobiologist Jill Banfield was puzzled by a pattern in the genes of bacteria that had been reassembled in her lab. A repetitive sequence of genetic code bookended stretches of DNA that matched DNA from bacteria-attacking viruses. Other researchers had dubbed the peculiar genetic pattern CRISPR (“clustered regularly interspaced short palindromic repeats”), and Banfield suspected that CRISPR could be an anti-viral defense system.
A professor in earth and planetary science and environmental science, policy, and management, Banfield looked online for fellow Berkeley faculty interested in RNA interference, used by plant and animal cells in their immune response. She located biochemist Jennifer Doudna, a professor of chemistry and molecular cell biology.
Doudna’s lab studied RNA, a chemical cousin of DNA, which is the molecule that codes for life. Banfield phoned her. Doudna didn’t know Banfield and had never heard of “crisper” but was intrigued by her colleague’s intensity. As Doudna recalls in the book A Crack in Creation, “… her excitement was palpable even over the phone, so I agreed to meet her for coffee the following week.”During their conservation at the Free Speech Movement Café, Doudna grew more curious about CRISPR’s unknown function. Her lab could design experiments to test Banfield’s hunch. “At the time,” says Doudna, “there was no experimental evidence that this was in fact an immunity pathway in bacteria, but Jill, with her incredible scientific sense, had the idea that this would be something very interesting for a biochemist like me to investigate.” Doudna followed her curiosity and started probing into the biology of CRISPR.
Some Dutch scientists even visited Yellowstone to look at Bacteria living in the Hot springs for CRISPR. The associate gene to CRISPR was termed CAS by Mojica.
Doudna was very interested to look at the molecular implications of CAS and CRISPR and wanted some solid associates at Berkely. When Blake Wiedenheft started studying microbes, his work was both remote and obscure.
He spent his PhD sampling hot springs in Yellowstone National Park, then created artificial versions in the laboratory to study the microorganisms that lived in the inhospitable water. "We wanted to understand how life could survive in boiling acid," he says.Over time, Wiedenheft became more interested in how microbes fend off viruses. He read around, and came across a peculiar bacterial immune system called CRISPR. In 2007, he approached Jennifer Doudna, a molecular biologist at the University of California, Berkeley, and found that she shared his interest. Wiedenheft demonstrated wit and persuasion showing Doudna all the contemporary research happening at CRISPR at that time including his tryst with researchers in Yellowstone. That is how he Joined the lab and revolutionized the field. Over the next five years, he studied the structure and biochemistry of CRISPR systems, landing a first-author publication in Nature. Wiendenheft used to cycle in the mornings as he was a great athlete could skii, wrestle , row etc. In the afternoons and evenings he was researching in Doudna's lab. Wiedenheft was mainly motivated by the crystal biology i.e how to map the CRISPR and CAS genes and see their folds, twists that would reveal much more than mere sequences as in Montana research. He was joined by Martin Jinek a czech scientist and they formed a great duo for Doudna , sharing humor and perspectives. Jinek and Wiedenheft were instrumental in setting up conferences in Berkelys the annual summits definitely helped in forging connections in the CRISPR community and advancing research. They both could painstakingly map CAS1 gene accurately . Meantime developments by french food scientist and biologist Barrangou helped advance gene editing he and his team first proved the Bacteria immune theory of Coonan and Mojica through a series of findings and later tried painstakingly to insert spaces into CAS genes of a Bacteria and making it 'Immune' to selected virus patterns through editing . This historic breakthrough paved the way for Gene editing revolution and solving virulent diseases by altering CAS. Barrangou was the first author on a paper published in Science providing experimental proof for the immune function of CRISPR, he later started collaborating with Doudna.
Doudna flirted with Industry outside Berkely wooed by the success of the firm Genentech which started in 1972 and pioneered lot of breakthrough in Scientific and pharma research such as insulin productivity. Doudna realized within 6 months into her stinct with Genentech that she was not cut for such politics and became clinically depressed. In 2008 /09 her chemistry chair neighbour brought her back into Berkely and Doudna started doing things which she did best research and motivate ppl to greater things. She was joined in this period by couple of smart PHD folks Rachel Haurawitz and Sam Sternberg . Together with an enormous appetite to change the world in CRISPR research Doudna , Wiedenheft, Sam, Jinek and Haurawitz made outstanding breakthrough in the CAS-9 molecular structure , thanks in no part to the whimsical molecular microscopic techniques that Sam Sternberg brought from Harvard to Berkely. CAS9 showed unlike other CAS genes to be able to memorize patterns in invading virus and alter patterns which was crucial to developing long lasting immunity . Again the folds and twists helped understand this phenomenon better. In this period Haurawitz, Sam attest to the mentorship of Doudna and her ability to challenge status quo and get them to do the absolute best work. Doudna also pushed hard at the scientific editors often arguing and getting good publications out of nature, science etc. The climate of collaboration at Berkely was never better than what it was at late 2009 . In this period Haurawitz due to her strong business acumen floated the idea of starting a CRISPR Start up based on CAS 6 advances . Caribou was born a mash up of CAS and Ribonuclei acid . Doudna was the chief scientific officer while Haurawitz the CEO and president. Doudna mentions how the venture capitalists even in 2011/12 were male dominated and it was hard for women scientists to flourish. Doudna, Haurawitz leaned on friends and families to raise a lot of capital.
Charpentier was a free spirit and extraordinary scientific minds of this generation. Hailing from a small parisian family she got inspired in school days as she kept strolling past the 'Center for communicable diseases' to one day work there. Her science minded parents encouraged her pursuits and Charpentier did post doc in France before taking up multiple research stincts in memphis and New york . Charpentier discovered she liked to pursue things with passion but move on to focus / refocus energy in newer research early on - she did not believe in long term relationships and as with any european scientist enjoyed cycling and hiking outdoors. Charpentier moved to europe first to univeristy of vienna and then Molecular infection institute at Sweden. She in 2011 discovered and suspected that the 'tracer Rna' gene had more to do with CAS9 precision in cutting the DNA of viruses but could not prove it experimentally . Taking her paper she reached out to Doudna and her team who hit it off with her immediately . Charpentier's post doc fellow chelinsky and Doudna's trusted Phd Jinek were assigned to conduct experiments on this , Jinek particularly jumped at this as he was having self doubts on his choice of field. In 1 year the foursome collaborating over skype made two remarkable discoveries a) Tracer RNA not only helped provide the DNA guide in the CRISPR RNA but also helped CAS9 in finding patterns and cutting the DNA of viruses. B) CRISPR RNA born out of Tracer RNA could be unified as one GUIDE RNA painstakingly. By June 2012 the foursome published a paper on their discoveries in Science . Parallely a team of scientists namely Barrangue and a Lithuanian scientist Virginijus Siksnys came up independently with much of the findings on CRISPR that Gene editing could be heralded by CAS9 RNA and CRISPR RNA .. However where they failed was the practical work and especially not understanding the full impact of Tracer RNA's role there. Siksnys paper unceremoniously and unfairly got rejected by CELL and Cell reports while Doudna's one found publication at Science. This infuriated Barrangue and he as the chair for the annual Berkely CRISPR conference put Siksnys as the first presenter on the topic before Doudna's team. Doudna made siksnys and barrangue understand that they got bested fair and square by asking questions and also demonstrating how 'Tracer RnA' is the most important molecule they failed to study. Charpentier after this remarkable discovery rather surprisingly decided to pursue microbes instead of Genetic editing as a career. Charpentier admits she being french knows how to ride the good wave but finds Doudna frenetically competitive and maybe that’s what landed them the accolades and recognition. In 2012 post their 2 strong discoveries doudna , charpentier found 3 tough challengers to take on . The CAS, CRISPR team had a major advantage to the other existing gene editing mechanisms which all involved using proteins / enzymes to cut DNAs . These proteins unlike the CRISPR RNA Guide could not be modified that easily and the instructions to target a DNA part was laborious and borderline risky.
The usage of CRISPR tools in human genome was a question mark that needed to be experimentally solved. Doudna's lab unfortunately did not specialize in Human cell biology and needed to bring expertise fast. Doudna challenge came in the form of George Church who politely told them via letter his team will be indulging in the experiments from her paper on human and Fang Zhang church's disciple who spectacularly fell out with him in Harvard. Feng Zhang had a typical bright immigrant story his mother was initially a visiting professor from Beijing to Massachussets and got enamoured of the US potential and stayed behind with H1B. His father learnt english after coming to US , Feng was 10 in 1991 when his mother became a professor at one of the colleges. His curiousity to 'make and do' things himself was nurtured by his mom and his passion got him into the des moines sting program for bright science kids. Funding in research was at an all time high in the late 90s and Feng capitalized on it. Feng went to harvard at the same time as Zuckerberg and he did some notable thesis work on inserting genes from jelly fish (that repel UV rays) into cancerous cells or those around showing it stopped cancer spread. Feng got good mentors in Levy who helped him see the Gene possibilities and Church who brought the best in him in the DNA / RNA space. In the 90s Gene therapy i.e inserting genes into organisms to cure genetic mutations was picking up steam but there were lot of side effects for this. Church felt feng was being sneaky in some of his papers and they parted way. Feng supposedly had two cores one ultra competitive and ambitious ,another humble and beatific. Doudna says just like how Kirik, Watson had Pauling , Newton had leibniz on calculas - She and Charpentier had Church & Feng to bring the best in them. Feng Zhang in 2012 with his team commenced work to show how CAS9 can be used in human genome. Church had a rather different upbringing with 3 step dads . Luckily for him one of the step dads was a hormonal surgeon who showed him some remarkable experiments and provided him chance to inject thyroid , pain killers as hormones to his patients. Church was a genial gentle giant a cross between Hagrid and Santa ! Church was the least competitive in his field and prided himself on integrity of the work to do good. In the 80s Church worked on radical experiments to sequence DNA and started few companies . In 2008 he became a household name in science for his pursuit of reengineering the wooly mammoth through the frozen hair fossils in the arctic and using the elephant's skin cells as embryonic gene . Idea was to unearth from this embryo using Gene editing techniques the Mammoth's sequence - Jurassic park style!
Feng Zhang preferred secrecy when it came to matters of competition (ultra some may say) . He was a late comer to the CRISP party and found out only by 2009 / 10 the possibilities of CAS & CRISPR enzymes as a superior tool for tool editing. Zhang had been working on Gene editing and cellular biology under Church (who was his mentor) at Harvard and always looked out for better toolsets.Feng joined the Broad institute under Eric Landers and pioneered Gene research at Broad. He met the Argentinian Bio researcher Luciano Marraffini who focused on CRISPR research to help him use CRISPR in mamalian cells, this collaboration was a great without being outstanding. Marrafini used many of Feng's radical ideas in his experiments at northwestern univ chicago in a bacterial cell. Marrafini pleaded Zheng through data to consider CAS9 as opposed to wasting time on other CAS genes . They made small incremental breakthroughs by planting NLS (Nuclei Localalization systems) that helped proteins to get into the Nuclei of cells and injected CAS9 time to time with minimal success for DNA editing. However their lack of full understanding of Tracer RNA and its implications in 'designing the Guide' and then targeting exact spots on the DNA meant they slogged much without real results to show for. A visiting Beijing Phd student who partnered with Feng lynn exposed Zheng's claim that they made most of the progress on CRISPR and found the mechanism before Doudna and Charpentier as false. Lynn says by June 2012 until Dounda, charpentier published the paper on Tracer RNA's use for cutting precise parts of the DNA via CAS . Feng and Maraffini had no idea and marrafini agreed to this. Feng , Landers and Broad university put up a vain fight on the matter and tried to cover their tracks by showing multiple experiment accounts but nothing conclusively proved they used Tracer RNA. Only by 2013 the extremely secretive Feng published a paper accounting for how Genome editing in mamallian cells can be carried out by CRISPR and unsurprisingly Marrafini was not sited as an author as they fell out. Lynn though featured.
Interestingly the June 2012 paper findings on unifying RNA into one simple molecule and the Tracer RNA role was mentioned. Church was another one who had a major tussle with Feng calling him unethical for stealing his best grad student Lixon for the CRISPR research without mentioning anything to him . Landers defended Feng and Broad faculty stating they did nothing wrong and Church's Narcolepsy (sleeping disorder) was as much to blame for this. Church was now extra motivated to beat the Broad team for their antics , Church argued to Isaacson given his reconciliatory nature Feng would have actually benefited greatly by Church's mentorship and maybe could have found a way to collaborate with Doudna's team. Doudna meanwhile after 2021 June smelled a large scale competition on CRISPR for Human genome and commenced research egging Jinek to figure out cellular biology ASAP. Charpentier largely stayed away focusing on other research. Doudna's ambition found luck in the form of Alexander yeast a fresh bio graduate coming from Feng's broad medical institute. Yeast applied many of Feng's techniques at Doudna's lab to grown human cells and then inject them successfully with CRISPR, CAS9 with DNA guides (Expected genome sequence) . The results astounded Doudna who again wanted to immediately publish these findings. Yeast was made to fine tune experiments and Doudna stated how even a graduate can run these experiments but her and Charpentier's invitrio finding in 2012 was the fundamental block for all of Gene editing research. Feng of course differed arguing cellular biology was superior to test tube research.
Doudna was proved right that her June 2012 paper proved genesis for cutting edge gene editing applications . Post her 2012 paper in December 2012 - Feng and then Church got Science publications for their Gene editing work in Human cells through CAS , CRISPR. Not only that by Jan 2013 including Doudna , Jinek publication there were 2 more 1 from another Harvard researcher and 1 from South korea all crediting her 2012 paper. Doudna can now claim the June 2012 test tube experiment is the mother of Gene editing. Doudna though fell out with Charpentier who increasingly became cold as she did not like Doudna sharing all the limelight for what she percieved her 'Original findings'. Charpentier started a company with her ex boyfriend Novak called CRISPR Therapeutics. Meanwhile Doudna tried hard in pursuing charpentier to join her plan of forging a CRISPR collaborators forum and eventually starting a big enterprise. Initially Novak , Charpentier came for these delegations in Boston but once it became clear Novak wanted to be CEO without due process and as others were opposed to it, they quit. One thing charpentier warned Doudna not to trust the 'Boston' gang who are all interconnected. Doudna went with Church, Feng and proceeded to commence a Gene editing company called 'Editus' however she was shocked to know that Boston gang (VCs, Broad researchers, Harvard + MIT) credited Feng as the CRISPR inventor and not her. Not only that Doudna recalls the biggest betrayal as the 'PATENT' done in Feng, Landers name for CRISPR,CAS when her Ip and that of Charpentier was cast aside and plagiarized. In April 2014 she quit Editus rather abruptly stating that they stabbed her in the back and preferred to severe ties with the Boston group. Doudna confided this in Church , who said all this was not a surprise to him and he decided to continue on . Science can be brutal and it is important to lawyer up . Charpentier , Douda's work did not by then get a patent and that proved their undoing as the VCs, Scientists pushed it in Feng's favor. Landers was also a big shareholder in several companies in Boston.
It becomes clear to Isaacson the reason charpentier and doudna decided to collaborate for the ultimate prize - nobel . After 2014 patent to Zheng , Doudna & Charpentier did get multiple big recognition including Gardner prize from king of Norway and 3 million (Siksnys finally got a deserved recognition).
Breakthrough Prize in Life Sciences the new nobel equivalent organized by Google , FB founders was awarded to the CRISPR crew but church was omitted . Doudna graciously donated 100K to his genetics research lab. The prize stated
"for harnessing an ancient mechanism of bacterial immunity into a powerful and general technology for editing genomes, with wide-ranging implications across biology and medicine." This showed despite the efforts of Broad to undermine Doudna and charpentier's 2012 paper the scientific and business world honored their breakthrough as one of a kind. In 2016, Lander published a paper in cell outlining the history , evolution of CRISPR research calling the 'Heroes of CRISPR' and prominently undermined and sidelined the contributions of Doudna to one para instead proclaiming the work of Zheng significantly. People thought this was another dirty trick to wipe out the annals of history particularly the 2012 paper and get Zheng ready for Nobel . Twitter storm broke out and Lander was widely labeled sinister and unprofessional. Church decreed the move as Broad's way of taking out Berkely's patent and Doudna was understandably furious . This feud boiled over to internet , political sides outside the universities. Lander and Zheng got their patent in 2014 by going via accelerated examination process that required more money and more diligence . Surprisingly despite 2012 paper doudna charpentier decided not to expedite patent quest and all that came back at them with a vengence.
The patent lawsuit first started when Doudna pressed the 'Interference clause' in 2015, whereby any other competing patent in the same period has a right to interfere with the patent claim. In 2017 a state judge bench overruled Doudna's interference and upheld the patent claim for 'First ever genetic editing in Human cell' of Zheng. However the breakthrough for Doudna and charpentier came once their patent got approved in 2018. Post this with a bright lawyer Alisson who also knew biology Doudna brought up the lawsuit fundamentally challenging possibility of Zheng's work without her CAS9 2012 paper and this prolonged in 2020 without clear verdict. Meanwhile in the period post 2015 there was frenetic breakthroughs and happenings in real world Gene editing first spurred by Charpentier's CRISPR thereapeutics - They successfully demonstrated how a stem cell gene can be edited and injected into an african american Gray in missisipi curing her by building gradually good blood cells to fight the 'Sickle disease' syndrome. Almost 100K african americans have sickle disease and precious little has been done on this . Given the high cost to cure 'Gray' almost 1 million the congress laid a challenge to public-private partnership to find a way at lower cost / affordable. Doudna jumped on this with her lab getting support from Govt and Gates foundation. Similarly new breakthroughs in cancer research was happening using the P53 gene which helps fight cancer in T cells in China . US had several regulatory roadblocks nonetheless thanks to the chinese it spurred a sputnik like US motivation and now Cancer was a prime topic of CRISPR Research. Apparently elephants don’t ever get cancer because elephants have extra copies of two cancer-fighting genes: P53, which hunts for cells with miscopied DNA, and LIF6, which obliterates the mutated cells before they can form a tumor. New research on other genetic disorders like Azheimers, male balding , edoema started . This also created private bio hackers to emerge.
Zayner was the prototype bio hacker . Having dropped out of Post doctoral research from University of chicago , he wanted to democratize CRISPR ensuring everyone had access to its goodness, Zayner even rejected work at NASA one point. Zayner started Odin a creative start up that provided things like Bacterial crispr kits for 169$, build your own genetic lab at 2000$ . Naturally the established research faculties called this foul play. However bio hacking found a surprising ally in Church . Church became the scientific advisor and mentor to Odin and Zayner . Zayner ran some radical research videos online DIY frog muscle therapy and a famous Gastric experiment. Thousands of social media followers lapped it up and the millennials started buying oden's kit to experiment. Zayner's vision was analogous to how US govt and military helped democratize internet , Personal computing, the Web etc. Meanwhile US military and DARPA was funding cutting edge CRISPR research at Harvard, MIT, Berkelely , Cold spring harbor etc. Doudna wanted to prevent misuse of crispr as a Bio war tool especially since Zheng published one of a series of papers illustrating how CAS9 can be programmed to cause cancer in lungs and other parts in Mice. Doudna's fight to establish a defense mechanism led to the formation of ANTI CRISPR proteins, which can stop rapid Gene editing and CAS9's efficancy. Unsurprisingly this find actually came from investigating virus and their evolution against CRISPR in bacteria. Genetics and ethics have long been a touchy battleground . With Aldous Huxley's brave new world showcasing how in future there maybe a superior genetically engineered human populace that controls the 'working population' and multiple other dystopian theories like the 'Fabricated man' published which famously alluded to the question 'Who can play god ? Man should first learn to be man before playing god' . Ethics in Genetics was widely discussed in 1973 by the DNA pioneers Kirik, watson , Berd in a conference where they proclaimed a stop to 'Recombitant DNA' research at a watershed 'Gordon conference'. However by late 70s and 80s it became apparent genetic research for other goodness like curing cancers, myriad disorders and virulent diseases was the way forward. In 2020 , it is highly probable according to Pentagon, DARPA that terrorists and malevolent state actors can easily modify genes for destructive purposes , this in the wake of the COVID epidemic is chilling.
The 1975 Asilomar conference was huge watershed moment for policy making in science Maxine Singer, Norton Zinder, Sydney Brenner, and Paul Berg were among the participants at the Asilomar Conference. The key proclamation from this conference was a) Genetic editing with Recombantant DNA i.e bringing dnas from different organisms into one gene shall not be used unless for diagnostic, curing diseases b) Policy makers / Government interference guidelines. This dawned an era for responsible research , james watson was an ardent opposer as he build scientists should be able to research however they want. Watson's argument was his schizophrenic son is a living proof that if humanity had powers to come up with 'Designer babies' then they should embrace it. With the 1978 in vitrio birth of Louis brown topics ranged from choice of gender, height, eye color, skin color, musical ability, physical / athletic prowess , repulsion to diseases etc . All the above took the form of the 1997 hit movie 'Gattaca' that showed how Ethan Hawke (a normal human) had to battle super human designer babies to become astronaught . Ebert in his critique threw open questions ranging from " Will humans be able leave to chance human conception if they really had choice to hand pick babies?" . However tampering with nature could throw in its own side effects as shown in 'Jurassic park' or bring about survival of fittest among super healthy long living race. In 1997 at Oveido spain the European countries except Britain and Germany took the stance that gene editing , gene therapy will only be used for curing diseases and preventing diseases not cross the 'Germ line' . This is the red line that helps to designer babies by choosing what to pass into the Gene inheritance. US did not go into the same ethical dilemma till 1999 when a swedish 18 year old boy
Jesse Gelsinger suffering from a rare liver genetic disorder took the audacious step of going into a trial gene therapy. Unfortunately he passed away as the virus used to pass the 'Good genes' engineered solicited severe immuno response leading to his lungs, kidneys organs failing one by one. This led to clamour and gene editing was effectively seen as Taboo in US despite the odd success of 'Dolly' the cloned sheep in 2003. Gelsinger was truly an altruistic heart as he told a friend 'what worse can happen through this , I may die .. But If I live I save a thousand plus babies :) '. Doudna recalls the backlash in 1999 and all these found a voice with the ultra conservative republican bush administration via the CAS commision who rebuked any attempt at innovation in Genetic research in that period. US had a genetic research vaccuum which was threatened to be occupied by china, russia till CRISPR came available. Everything changed and by 2011 Sternberg under Doudna even contemplated working with Buckman on what was a controversial topic 'Healthybabies.com'. Buckman came from a noble intent having seen her sister suffer through leukemia. Though this project did not materialise due to the several logistical, policy nightmare they have to navigate through.
He Jiankui was a story of the talented immigrant from china making big , except it was not . He was a very bright promising post grad (PHd) student at Rice on Genetic engineering. Professors including Michael dean gush how much brain power and extra curricular passion He jiankui had as he demonstrated often in the 6 Rice soccer fields. Jiankui went to stanford for research and picked up CRISPR CAS9 editing as his main thesis and subject. Soon thanks to a wealthy VC in Shenzhen , Jiankui started his own Genetics company pioneering Gene sequencing in China for a sum of 100$. Jiankui did not stop there after 2017 with multiple breakthroughs coming everywhere in the world on CRISPR edits. Jiankui decided to engineer babies that are disease resistant and can come out healthy. He wanted to rid the stigma AIDS couples were facing and took up that as a personal ambition. Jiankui however was very committed to not using Genetic edits for enhancements, skin color, physical prowess etc. He got 15 couples (with one or both of them HIV positive) register on his ambitious project to produce HIV resistant babies. Through sperm washing techniques editing in the CCPR5 gene finally Jiankui in 2018 November did the unthinkable as his patients Grace and Jack concieved a twin girls. Nature did not accept or publish his paper slated for Jan 2019 as this landed a huge controversy amidst the scientific community. The templeton foundation which was established in 2017 with Doudna as one of its chair found Jiankui adamant and not caring enough about ethical implications of Gene editing. Jiankui felt the world will never be ready until some breakthrough arrives showing why CRISPR needs to be more maintstream. In the 2019 Hong kong conference most researchers including leading chinese researchers took exception to the meddling of Jiankui since this approach was not well tested before applying to humans. Jiankui ploughed along but found even the chinese daily mouthpiece retracting his achievements. In the face of this storm that he seeked counsel with Doudna .
His 2019 Youtube videos on the whole experiment leading to birth of Lulu and Nana stirred the hornet's nest everywhere. Michael Deem his Rice supervisor went under intense scrutiny in US for his aid of Jiankui's efforts , Although Deem denied wrong doing video and other evidence incriminated him strongly. More clouds of disgust circulated with the rumor that those two twin girls may have had brain enhancements as a consequence of the embryo building. Jiankui kept putting off these accusations of 'Mad scientist in wild wild east (china)' saying it was inhuman if humans did not intervene knowing a fetus would suffer diseases.
Doudna , Baltimore and others in the Hong kong conference while decrying jiankui's madness in not following protocol or best practices did not call for a moratorium on what is going to be the future of humankind - Germ edits. Unsurprisingly it was Doudna's arch enemy Eric Landers that called for a widespread moratorium on this. However the EU court of Justice in Sept 2020 said its imperative science forges ahead with ethical checks and balances on the Germ editing phenomenon. This was in light of the enormous health strain placed in the wake of the Covid 19 pandemic outbursts. MRNA vaccine thus became common place and largely a success story of science. Ethical conundrum sprung when two deaf lesbian couple in midwest USA went searching for a sperm donor who was congenitally deaf so they can have a deaf offspring for certainty and they got one. This sparked discussion should humans have options to edit out 'Hearing' in genomes if they want to , what about puncturing ear drums on birth ? Is it humane to birth babies with defects just because parents or society wants such variation ? Is it justified on the babies knowing they could have been healthier if not for the wishful tampering. George Church did not mind using genetics for enhancing IQ points or height , he believed if there is capability our future generations deserve better in fact best . However on deeper thoughts opening up a Genetic maketplace that is not well thought out with gates can be a catastrophe - People may be biased to choose 'Straight ness' for conformity and of course evolutionary conviniences , extra muscles, IQ and things can veer into blonde eyes and white clear skin (Ring familiar to the Nazi , Eugenics era). Darwin argues there is no benevolent or good designer up there as evolution scrambles and picks things on by necessity . In fact there are plenty of bad designs continuing in species like the poor sinuses of apes, bad urinary tracts for men etc. However this chance and random evolution is still the best bet to prevent designer babies and the super rich doctoring and skewing society to their benefits. If at all we decide on producing designer babies through choice of genes - What would be the goal ? Eliminating crimes , Blissful existence ? Impervious to viruses? Prolonged life ? Achieving success by 'Experiences' on own ? Its hard to quantify and with slight variance of the goal posts the world alters dramatically. One thing most scientists agree and feel hitherto needed is the elimination of possibility for genetic disorders totally in babies - Huntingtons, sickle cell, even predisposition to cancer and diabetes. Brave new world and Huxley show distopian future of Alpha babies (rulers) and Epsilon babies ( menial labor) who are given Soma - to keep in bliss. One more huxley imagination shows an island where couples volunteer to be impregnated by men with superior IQ, physique and less disease possibilities to keep randomness to minimal. 1984 argues how totalitarian regimes can completely eliminate creativity and individual think tank by a single minded order. However in today's hyper information age the problem is not from these isolated dictators esp in the west but lack of unregulated media, social media that can create conspiracy, false information in micro seconds. We are not far away from Homo sapiens 2.0 - Super soldiers who are tolerant to Nuclear attack, night vision , endurance without sleep etc. This would mean like software version Germ edit version of genes may come every month as an enhancement . Imagine like Iphone versions new versions of these germ edits are given every quarter. But that also gives room for malevolent actors tampering genomes and bringing down populations through bio attacks. Imagine dating , employment, military using genes as a way to determine source of truth. Most researchers in the Quebec conference on CRISPR including Feng agreed that Gene editing could become dangerous if not solved well and thoroughly ethically. Rich people had better access to everything skewing populace in their favor from health care, technology, education , quality of life etc. It wont be any different if left to the 'individual's choice' in the long run . Already Bezos, Thiel are looking to live beyond 120 with anti aging research and they would jump into germ edited enhancements.
Isaacson meantime could validate first hand that doing edits via CRISPR CAS research was easier than expected he went to Zurich university labs of Gavin Knott and Hamilton and did things in pipettes himself. He exclaims how easy if unregulated any freshman , rookie or worse bio hacker esp malevolent ones can do with the tech ? James watson at cold spring harbor meantime worked himself to a corner for more distateful eugenics odour - watson repeatedly spoke how blacks are inferior genetically and got himself into a soup . Cold spring harbor unfortunately had to let him go and removed all traces of his distinguished contributions from their univ including a huge portrait and his honorary titles. Watson though was eccentric progressive in one way esp to solve cancer, mental health issues genetically like his son Roofus. On the other hand believed nature not nurture was reason for black impoverishment , African lack of growth etc. Doudna , Landers sympathised with him as a gentleman unable to step out of the old world. Landers given his earlier faux pas with Doudna on the CRISPR contributions got himself a hot potato by toasting for Watson as the 'Double helix' man. Watson firmly believed everything on 'Genes' which did not go well in the real world. Isaacson met Watson in person and could feel he regretted his direct comments but not his beliefs . When prompted to talk about race he was still firm in his thoughts , Doudna felt we should accept people for their eccentricities similar to her dad Martin doudna. No one is perfect . Isaacson found Roofus , watson's schizophrenic son very protective of his dad and Roofus even felt his dad deserved a better smarter kid than him. Isaacson wonders how much fall from grace Watson has experienced , going back in late nineties he was one of the time person of the century and now in 2019 was entirely bad odor to be ostracized. Whatever the repercussions currently it will be an insult to history if Watson is not credited his rightful place in Genetics for his Double helix findings that started a new revolution.
Vaccines - MRNA vaccines started with Moderna and parallely Pfizer/ Biontech commenced this. Moderna and its armenian founder wanted to explore mRNA to solve cancers and when 2020 COVID virus broke both companies raced ahead from get go. mRNA vaccines inject a virus with the CRISPR/ CAS9 into the cytoplasm i.e provide the code of the virus the proteins have to be produced accordingly. Traditional vaccines including the famous ones for small pox took a weakened virus or dead virus and inject antibodies from another animal , in the case of smallpox it was cowpox virus to generate anti bodies that works against small pox. Essentially vaccines help body to produce anti bodies. However unlike tradional vaccines that shoot in the dark with templates - Genetic / mRNA vaccines give you the exact code of the virus that is going to be very relevant to what we are fighting against. Numerous modifications, mutations can be covered by the high level code and it is very specific.
CAS 13 was a huge find that accelerate another couple of corporations to find a virus and cleave it up mercilessly basically not waiting for human body to generate immune response. This was trickly to launch into Lung cells where COVID affected and hence it could not take off till 2021. Cameron Mhyrvold was a pioneer of this at Zheng's broad using CAS - 13 A, B which could detect in tandem with sherlock and then cleave out virus. Meanwhile from Doudna's lab CAS-13 D a superior targeting RNA was found to give better results. Two technological outputs emerged with this called CARAVER and PACMAN to cleave virus it is not used widespread in medicines or vaccines just yet due to some unchartered tests missing like launching and any side effects.
Meanwhile the book concludes stating how CRISPR sparked a revolution that reached fever pitch due to the seismic activity of COVID and now every household has testing kits which is a milestone for gene engineering. Doudna, Charpentier were pleasantly surprised when the Nobel for medicine came to them in 2020 in the middle of the patent suit . Zhang , Church were notable misses but Eric landers did praise their 'Contributions' highlighting yet again how women are prominent in research. Doudna, charpentier never really patched up and did share few cozy moments. Doudna was still hurting at missing her friend of several years . Isaacson heralds with this book how Genetic engineering is the next big revolution like Tech was in 2000s and cloud computing / ML in 2010s.
