Introduction
- Response to coronavirus
- “She lead by listening.”
- Proposed ten projects for her lab to take on (testing labs, diagnostics)
There’s a joy that springs from fathoming how something works, especially when that something is ourselves.
- Some themes/goals of this book:
- Nature is beautiful
- Impact that a persistent, curious, and competitive individual can have
- Other star players in field of gene editing, but Doudna’s instincts of collaboration with a competitive streak make her good choice to study
- How Doudna’s life story connects to larger historical themes like women in science
- Key to innovation is connecting curiosity about basic science with practical work of devising usable tools (like da Vinci)
- How science actually works
- To reiterate: importance of basic science and curiosity-driven research
Part 1: The Origins of Life
The key to true curiosity is pausing to ponder the causes.
- Discovery of the genetic code coincided with information revolution (digital coding) → “The flow of history is accelerated when two rivers converge.”
- Grad work under Jack Szostak at Harvard… several key lessons:
- Never do what a thousand other people are doing (focus on RNA despite hype about DNA)
- Ask big questions (origins of life?)
- Worked on ribozymes, proving that RNA can self-replicate
- Post-doc with Tom Cech at UC Boulder; studying structural biology & self-splicing introns
- Moved to Berkeley → how does viral RNA (like that of coronavirus!) interfere w protein-making machinery?
- Studied structure of Dicer; showed how part could be reengineered to use RNAi to turn off other genes
Part 2: CRISPR
- People initially though CRISPR worked through RNAi → how Doudna eventually got looped in
- New post-doc Mark Wiedenheft worked on figuring out fx of Cas enzymes— focused on Cas1, when only one found in all bacteria w CRISPR systems
- Vannevar Bush → linear model of innovation: basic, curiosity-driven science leads to technological progress
- Led to founding of NSF to fund basic research
- Some truth to this model, but business interests, engineers, etc. also essential to progress
- In CRISPR story: yogurt makers at Danisco got interested bc wanted to protect its bacterial cultures from infection
- Studied sequences from samples they had saved over time → understood mechanism, started “vaccinating” their bacterial strains with CRISPR (2007)
- 2008 → first CRISPR conference, discovery that the system works by targeting DNA, not RNA → potential as a gene-editing tool
- Two components of scientific discovery: “doing great research and building a lab that does great research”
- 2009, after 2 month stint at Genentech, Doudna returned to Berkeley, began focusing on lab over benchwork
- Relatively hands-off at beginning of projects, but asks the right critical questions among the way, engages more as they get closer to fruition
- Doudna lab looked to understand structure & function of other Cas proteins → “unexpected recognition mechanism” in Cas6
- Biotech industry followed tech trend (which began mainly at Stanford) of blurring lines between academia and business, scientists & VCs teaming up to commercialize discoveries, profs remain as faculty
- Model Doudna followed to bring research to industry — first with Caribou (Cas9-based diagnostics)
- More recently, philanthropic foundations join academic-government-business ventures, too
- Emmanuelle Charpentier → “found in biological research the right combination of persistence and creativity”
- Worked on tracrRNA → facilitated making crRNA (which contains viral seq), scaffold to attach to Cas9
- 2010: dIscovered that tracrRNA, crRNA, and Cas9 → fully functional defense system, but still unsure what tracrRNA did after processing crRNA (important to ensuing IP drama)
- Worked with Doudna to elucidate mechanisms along with Martin Jinek and Krzysztof Chylinski → found that tracrRNA was req for crRNA to bind to Cas9 (in test-tube rxn)
- Discovery of programming capability of Cas9 → Doudna et al. engineerined minimal tracrRNA → sgRNA (significant bc human invention, 2012 Science paper)
Part 3: Gene Editing
- Would CRISPR work in human cells? And was proving so a separate inventive step to what Doudna et al. had just shown?
- Feng Zhang → exceedingly humble, gracious— and competitive
- At Harvard at the same time as Mark Zuckerberg— interesting to consider who will end up having more of an impact on the world (in general, relative importance of biotech vs digital tech)?
- Sensed that CRISPR was going to be a big deal, and kept his work on it v quiet
- Difficulty sorting out essential Mike uses of CRISPR-Cas system, vs Doudna who had expertise with that but not with working in human cells— worked on nuclear localization, optimizing sgRNA, etc.
- Controversy over how important Doudna and Charpentier’s work, esp re: tracrRNA, was to his work in editing human cells
- George Church → “a gentle giant and a mad scientist,” mentored both Doudna and Zheng
- Reached out to Doudna after CRISPR paper was published a interest in trying in HSCs… but didn’t talk to Zhang bc didn’t know he was working on it
- More inclined to cooperation and openness than competition and secrecy
- Without knowing it, submitted his CRISPR human cell editing paper to Science within a day of Zhang → some temporary tension btw the two
- Doudna and other’s papers applying CRISPR systems followed shortly— inevitable once it had been proven in vitro?
- Eric Lander (at the Broad w Feng Zhang) wrote 2013 article on the history of CRISPR, downplaying the role of Doudna and Charpentier → lots of backlash from media, other scientists
- Doudna applied for patent first, but Zhang had his expedited and was granted → conflict!
Part 4: CRISPR in Action
- Current applications = treating sickle cell, cancer (ex. CAR-T), blindness, diagonostics
- Affordability is key concern
- DARPA → funding efforts to defend vs potential weaponization of CRISPR
- Discovery of anti-CRISPR mechanisms in viruses
Part 5: Public Scientist
- 1975 Asilomar conference, led by David Baltimore, → scientists trying to regulate themselves re: recombinant DNA → lift moratorium on gen eng, proceed with safeguards
- Did not address question of how far to go in editing human cells, if/when tech was achieved
- Eventually general consensus in Eur = against germline engineering, both in agriculture and humans; less opposition in America
- Death of Jesse Gelsinger following gene therapy → became controversial
- Doudna started getting concerned about potential misuse of CRISPR stuff, became more engaged in public policy about gene editing
- 2015 conference in Napa → ethics of naming inheritable genetic edits
- General consensus that a hard ban wasn’t a good idea; rather should set out guidelines for safe research
- Agreed on “temporary halt on germline editing until safer and social issues could be further understood”
- Language and position of the stance continued to shift, varied by country
Part 6: CRISPR Babies
- The whole thing with He Jiankui, 2018
- Investigations into his mentors in the States, most of whom discouraged him from proceeding but not much else
- Jiankui outlined five ethical principals he believed should guide future of germline editing, seemed to believe in ethical value of his actions
- Unfortunately, editing didn’t go perfectly → off target effects, only homozygous for one twin
- Along w Chinese media, thought he was going to be hailed as a hero when news broke…
- Despite uproar, official statement by Doudna, Baltimore et al was very restrained → germline editing no longer a red line
- Stronger call for moratorium by Eric Lander and the Broad
Part 7: The Moral Questions
- The continuum conundrum: is there even a line to draw, a way to separate categories?
- Germline editing is a pretty clear line, though not perfectly so— and that doesn’t mean it should never be crossed
- Question becomes: which cases, if any, should cause us to cross the germline?
- Much more blurry line exists btw editing for the goal of treatment vs enhancement — could *distinguish by defining whether an advantage is *positional (helps you by giving you an edge over others, like being taller) or absolute (would be helpful even if everyone else got it too, like improved immunity)
- Germline editing → offers a way to eliminate “middle-age” diseases not acted on by natural selection
- How to define what is or isn’t a disability? Consider autism, deafness
- Eliminating disabilities (even very clearcut ones) may make world less diverse and creative—but does that give the govt the right to prevent families from using gene editing tech to prevent suffering in their children?
- Consider sports— most runners have certain allele of ACTN3 → would ensuring your kid has this gene disqualify them from competing? Where to draw the line between natural and not?
- Who should decide? As with most moral issues, comes down to collective vs individual…
- Danger of enshrining inequality into our species if left up to individuals
- Doudna → cautiously in favor of germline editing, in cases of diseases like Huntington’s
Part 8: Dispatches from the Front
- Feng Zhang: “In a world in which there are people who don’t get access to eyeglasses, it’s hard to imagine how we will find a way to have equal access to gene enhancements. Imagine what that will do to our species.”
Part 9: Coronavirus
- Doudna = “maestro collaborator”
- Lab organized major testing facility
- When virus first started spreading in US, lots of university & hospital labs got together testing capabilities, but hindered at first by tons of FDA paperwork and bureaucracy
- Doudna’s lab (Mammoth) and Zhang’s lab (Sherlock) → Cas12 and Cas13 respectively as detection systems
- Both enzymes have property of indiscriminate cutting after finding a match
- Allow detection of RNA instead of proteins like most antigen tests (can catch cases earlier)
- Still driven by competition, but also openly sharing data and methods
- Some Doudna lab grads worked on Moderna vaccine
- Doudna and Zhang labs also both working on Cas13-based virus treatments (PAC-MAN abs CARVER)
- Challenge = felivery
- Advantage = ease of reprogramming
One fundamental aspect of science will remain the same. It has always been a collaboration across the generations, from Darwin and Mendel to Watson and Crick to Doudna and Charpentier.