Pushing the Boundaries of Synthetic Biology (Emily Leproust, Twist Bioscience) | iGEM 2024 (Keynote)

Good morning, thank you very much. Such a great pleasure. I go to a lot of conferences and IMM is absolutely my favorite conference, and it's my pleasure to share what we do at Twist. Really, what's super exciting is that our view is that DNA is changing the world. There are so many things that can be done with DNA. You walk around here and see all that excitement. Our customers are using the power of DNA to develop the production of chemicals using biofactories in a way that's more sustainable. Sustainability is great, but it's also cheaper. I talked to a lot of investors, and if sustainability alone is not enough, you need to also be cheaper. We have a challenge of food production: the number of people is growing and the amount of land available is shrinking, so we need to produce better food, and again, DNA technology is a great approach to do that. In health, both diagnostics and therapeutics — finding disease and curing disease — there's lots that DNA can do. And last but not least, as we create big data sets, the storage of data is becoming a problem for the planet, where data centers are using a lot of energy. Leveraging DNA will be able to store and archive data in a way that does not require a lot of power. So at Twist, we're here to help, and we've built a technology to do that. Maybe you know, maybe you don't, but usually DNA synthesis is done in 96-well plates, so you can make 96 oligos at the same time. What we've done at Twist is we've built a silicon chip the same size as a 96-well plate, but instead of making 96 oligos, we can make 1 million oligos. So we're not 10 times better, we're 10,000 times better. On that chip, we can make all the products, all the customers, all the applications at the same time. Because we need to track what happens where, we've also invested in a huge software infrastructure to track everything. Twist is that combination of the silicon chip, the chemistry, the biology, and the software. Now we are a thousand Twisters, and the pace of innovation is very fast. We invest about $120 million a year in R&D, which is why we're on a pace of innovation that's still very, very fast. As I mentioned, with the silicon platform, we have miniaturized the chemistry, so that lowers the cost and increases the scale. In a second, I'll show the impact on our ESG impact. But even though it's a great technology, at the end of the day, we are a business of people, and you can't do anything without great people. We've built a great team. It's a very diverse team: 61% of people at Twist identify as people of color, and 40% of our leadership is female. Biosecurity is something that's very important. As you know, DNA is very powerful, but it's also a dual-use technology. It's kind of like dynamite: you can make tunnels, but you can kill people. So we invest a lot in biosecurity to make sure we promote the ethical use of DNA. We screen people, we screen the customer, and we screen the DNA. If you order Ebola but the customer is the CDC developing a diagnostic test, we will do it. But if someone orders polio to be sent to North Korea, we won't. It's really that combination of screening people, the customer, and the DNA. We try to spread those ideas and the software for free to other companies so that everybody screens. I think it's very important that every company should be screening to make sure there's no accident or any misuse of DNA.

In terms of the products we have at Twist, I'll just talk a little bit about a slice of what we do. We have NGS tools, but I won't talk about that today. Today I'll focus on our synthetic biology tools. In the next few slides, I'll talk about Express Gene, our multiplex gene fragments, and our libraries. There's a lot of other things we can do, but just to give you a flavor of some of the applications and the technology we have. First, speaking about fragment quality, it's very, very important. Here we're showing the quality of our double-stranded DNA fragments. These are non-clonal, and as you can see around here, even for 1.8 kb, we have errors of less than one in 5,000. That's extremely high quality, which means that when you do your screening, you can really trust the data. Quality is super important. Because we use fewer chemicals, we have less of a carbon impact on the climate. We try to quantify what the carbon emission of making one gene is. When we do it the Twist way, the carbon emission of making one gene is the same as charging 4.4 smartphones, while the standard approach is the equivalent of charging 2,798 phones. Because we miniaturize the chemistry, there is a lot less carbon emission. So when you get the gene from Twist, not only is it high quality, fast, and cheap, but it's also good for the planet. Speaking of speed, we recently launched Express Genes, which means you can get a clonal gene in your vector shipped in 5 days. This is the production data from June this year, with several tens of thousands of genes, and you can see that almost 98% of the genes were delivered in 5 to 7 days. You can order 1, 10, 100, or thousands — you can't break us. We have huge speed, and for many applications, the design-build-test cycle needs to be as fast as possible. Two applications in particular: antibody engineering — if you get the genes in 5 days, you can get the IgG in 12 days. So if you're doing antibody engineering, the reformatting of IgG is critical. You can either get the DNA in 5 days or get the IgG from us in 13. At the bottom, we're getting a lot of customers asking us to make the DNA and then make the RNA. There's a lot of application around personalized cancer vaccines. You can make the DNA in 5 days and then the RNA in 2 more days with in vitro transcription, capping, and purification. Switching gear to ease of use and low cost. I mentioned earlier about Express Genes with your gene in your vector. We have a new vector onboarding tool where it's very simple: you drop your file, answer three questions, choose your insertion point, and that's it. You just send the tube to us, and we'll upload your vector into our system, and it becomes your custom vector. Then you can apply that to the full suite of products we have. It's a busy slide, but the one thing to see is we have two formats: one format is arrayed DNA, so one sequence per tube, and then we have a format where we have pooled DNA. So far, I've talked about the fragment that can go up to 5.2 kb, clonal Express Genes that can be delivered in 5 to 7 days. Next, I'll talk about something that sits between the fragment and the oligo pools: the multiplex gene fragment, which is a direct synthesis of 500 base pairs delivered as a pool. The benefit of those multiplex gene fragments is that there are a lot of great applications of oligo pools. At the top, you can use oligo pools to optimize antibodies by varying the CDR1, CDR2, CDR3 regions and the template. Another is massively parallel reporter assays where you can test thousands or hundreds of thousands of promoters to learn the rules. At the bottom, you can have ultra-complex CRISPR screens where instead of just making one guide RNA, you can put multiple ones to either do multiple hits on the same gene to ensure silencing is effective, or have guides that remove part of the fragment. There's lots of great things you can do. Encoding all that information takes length, and typically oligo pools are 300 base pairs, so it's not quite long enough. What we've done is increase the size to 500 base pairs. Now with that 500 base pair direct synthesis delivered in the pool format, all those applications are very accessible. You can make a full-length synthesis of a heavy chain or light chain, encode up to 4-5 guide RNAs together. This enables massively parallel assays that are very, very powerful. What's great too is the cost. Here is a cost analysis: if you add more and more fragments, you can go up to several hundred thousand. If you buy one fragment per tube, the price doesn't really change — not a lot of savings as you add more. But with the multiplex fragment, the more you add, the more you save. In science, more shots on goal is better, and now it's affordable. When we developed it, we'll show you some data. Quality is paramount, and it's very hard to make direct synthesis of 500 bases. We used our big R&D budget to push the boundary. We tried many recipes. This is a recipe where the 500 is good but you get a lot of short DNA. We've tried thousands of chemical recipes, and at the end we focused on this one, where you get no small fragments and 90% of the molecules are full-length DNA. The other thing is uniformity. You need good error rate, but also great uniformity. We sequence all our pools, and we want to make sure the ratio between the least and highest amount of molecule is low. Typically, 3x is a great uniformity, great quality library, and that's what we're able to provide. Even as we look at the distribution of length and GC, the uniformity is really, really high. Moving quickly to complexity. As I mentioned earlier, more oligos is better, so we have different types of libraries that can go to billions of mutant diversity. I'll focus on just one for the interest of time. The clonal variant libraries are very, very powerful because you can have multiple domains: CDR1, CDR2, CDR3, multiple domains of your enzyme. On our silicon chip, we can make millions of mutants. You can really control the mutants — there's nothing random. You can have base-by-base precision on which mutants are in there, control codon usage, length of the domain or CDR length, avoid restriction sites, remove things you don't like. At the end, we always sequence the library before you get it by next-generation sequencing, so it's very powerful. These are just some examples of applications our customers are doing with those pools: enzyme engineering, metabolic engineering, antibody engineering, cell and gene therapy. We're seeing a lot of great science with those pools.

In the last few minutes, I just want to switch gears a little and talk about IMM. We really like IMM. This year, we supported 50 teams; we provided free DNA to those teams so they could do their work quickly. My personal motto is 'friends don't let friends clone,' so the IMM team didn't have to clone. We shipped for free more than half a million bases — a quarter of an E. coli basically — to the teams, and they did great work with it. Since we're in the numbers, we calculated the number of brain cells in this room. It's a very big number, almost as big as the size of the libraries we can make. We can make 10 to the 15 libraries. The organizers asked me to share what drove Twist and what made Twist successful. The first thing — sorry, the slide deck is blocked. Can someone help me with the next slide? The first thing we focused on is sustainability. It's in our company tagline: 'DNA for health and sustainability.' Sustainability is such an important part of how we can make the world a better place, but as I mentioned, it's not sustainability alone; it has to be sustainability as part of reducing the cost of something. I'm going to try to click... Oh, now it's working. The next thing that was very important for Twist is commercialization. It's not only important to have a technology idea. If you want to be a startup, you really have to make it into a business. It's hard, but it's very key for the synthetic biology industry as a whole to succeed. We want to make sure it turns into a business. Thinking about how you will get your product or services in front of customers is very, very key, and you have to think about it upfront. The last thing that was very key to Twist is around legal and ethics. The IP strategy is very important; you need to make sure that if you have a great idea, you need to defend it. Thinking through the IP strategy early is key. Ethics is also so important because again, it's a very powerful technology, but we need to be able to make sure it's used in an ethical manner. To conclude, I think together we all have a shared vision of building a better world. As young people, you have an amazing opportunity to build the world you want to live in. There are a lot of challenges in the world; climate is one of them, but as a generation, we have an opportunity to do something about it. What I've seen here really shows that very powerfully. My last slide is really 'be one of the great.' I think in this room, we'll have someone that makes cancer a chronic disease, people who are going to have massive opportunities to make the world a better place. It's such a pleasure to be part of that, to be part of helping you. I can't wait to see what you are going to accomplish, and I can't wait to put your picture on my slides in the future. Thank you so much.