 Welcome to the English translation stream for genome editing with CRISPR in CAS, a new hope or attack of the clones. That's the question today. You probably heard about the research information about a Chinese man called He who edited something with the babies. We'll hear more about this later. The main tool that was used in that research or what is proposed to be research is a tool called CRISPR which is the scissors and another tool called CAS which is kind of a gluey thing. This talk is both an introduction and an outlook really respective to the how this is going to affect us, what are the dangers, what are the benefits of this research. Please welcome the speakers. Hi this is Catherine, this is Anna, I'm Andre. We want to talk about genome editing using CRISPR and CAS. Will this be a new hope or attack of the clones? What scissors? There was a lot of things in the international press. One is planned so genetically edited one being over design, the final taboo in Genetag to 290% of revenue using CRISPR-theored products. This is how it's going to continue. The battle about the gene scissors is just starting. And it's talking about patent attorneys, the bad guys. Exactly how this works is something that Anna will tell us about. I will give you a very short introduction into DNA, DNA the code inside our cells and that we use to survive. DNA is being translated or transported via RNA. Always three bases are coding for one amino acid. So if I change a base, the amino acid will change and proteins are made out of many many amino acids. And most things happening inside the cell are based upon proteins acting and the diagram in the bottom right is a codon diagram to tell you how with just four bases we can encode 20 different amino acids. So always three bases make up one, encode one amino acid and the language has four different ones to choose from. And what exactly is CRISPR now? CRISPR is part of the bacterial immune system if you want to call it that. The authors are the ones that have the patent currently. The images of the Ocola bacterium typically found in human intestines and these bacterias can themselves also be infected by viruses and become ill. So the virus attacks and dumps its own genes inside and this infects the bacteria which in turn produces more viruses. And this ultimately kills the bacterium and produces a lot of viruses which in turn can infect further bacterial cells. So what's now the use for CRISPR? When the phage attacks, the RNA will break apart and CRISPR will glue the code into the original bacteria code. This allows a second enzyme to later detect and destroy the bacterial RNA. This in turn allows the Ocola bacteria to be saved from the attacking virus. CRISPR only cuts at a triplet called NGG and this triplet is very prevalent everywhere in the genome. So it could in theory cut everywhere so there's another 20 bases to allow a detection sequence to allow for specific cutting. So application. This is an animal cell. We have prepared the DNA with a cast sequence which means the DNA is split apart and we now use the repair mechanisms of the cell because DNA breaking is very common in a cell. This breaking is being detected and then fixed either perfectly using kind of a glue or additional bases may be introduced. So that would be an insertion and an indel would be something that gives an offset of a different value than three bases which means the entire readout is now changed. Of course it can also be that more bases are deleted so this would be equally destroying the way it's being read. So the entire gene is being destroyed because it can no longer function to encode for the same protein. So how is this used inside a laboratory? We want to force cells to produce something they would not originally do or we want to make them produce something they already produce in a different way or we want to change structural parts within the DNA so not encoding for proteins but something else. So either introducing sequences or by moving the way it's being read out so that the code on structure. So basic research means we change something we look what happens. Before we get to more concrete information about how it's applied we want to make some comparisons to technology. So Cas9 in the bacterial immune system is basically a virus scanner using GRNA as a signature. There's also a comparison with a text editor or word or of course the gene scissors. The connotation is this is precise and it's correct technically but this is a biochemical system and not a binary system so it's not always that easy because for example DNA and RNA molecules that don't fit 100% but almost can adhere so the Cas9 may cut in places that weren't targeted. So a good comparison is this sort of broken reg X which would cause off target effects and side effects and side effects have associated risks. So back to Anna. But people have thought about how to minimize these off target effects. Cas9 has a certain error tolerance but you can use bio tools of bioinformatics to find sequences that are as unique as possible. Or you can use Cas9 Nicas which needs two guide RNAs. So your sequence size is now 40 bases and you have a much lower probability of off target effects. Then there are different Cas proteins that have a different specificity when cutting and of course anti CRISPR proteins which deactivate CRISPR. That's the basics more or less. CRISPR makes basic research a lot faster and a lot more precise. But now CRISPR in medicine and the big question is can we cure genetic diseases? So yes if the disease is monogenetic meaning it's only caused by one gene. If it's often the same mutation causing it because then you can just reuse a system you already established. Gene doping doesn't work because the effects usually involve many genes so it's very difficult to handle and it's not really practicable. The question is how do we get CRISPR cast into the cell? For example we can extract stem cells, treat them externally and put them back in. But of course you can use a virus as a vector but that's one of the greater difficulties. There are studies about 20 of them. One thing is creation of CAR T cells against cancer. Those are immune cells that are extracted from the patient rewritten and put back in. They do it already but you could also do the modification with CRISPR which would be much faster and cheaper and also better. Then there's beta thalassemia or HPV and like an HPV cutting out viral sequences. Then there's this one about phenylketonuria which seems to have been cured. Thing is if you read a bit more it has been done in the mouse model and the disease involves a gene being broken. Thing is there are more than 850 possible mutations causing the actual illness. So now they fixed one but there may be more. This interesting and research should be more clear in communicating which model they used. So now the really interesting part editing inherited DNA directly. Like an embryo sperm, an ovum. In that case the change you induce via CRISPR will be in almost all cells of the resulting being including all germline cells. So it would be persistent. The potential is you could genetically fix inheritable diseases and the human in question would be born healthy. The success quota in embryo tests is right now around 70%. So now the question before we get to the ethics of it, is it useful? Because we have every gene doubled. So if we pass it on, we pass on one of both, the other partner transmits another one. So there may be embryos that are naturally healthy even though both parents are ill. So that might be less controversial is a selection of embryos through prenatal screening. You don't do anything different with CRISPR. You have to sort through the edited ones and junk those where the edit didn't work properly. There are ethical properties, throwing away the failed ones. And with CRISPR you have off-target effects and lack of consent of the treated individual. So almost the whole world has past a moratorium on the thing. And the UNESCO is trying to establish a global standard. There are some biohackers who put a lot of hope in CRISPR. And the best thing that can happen is nothing, a failure. Or you can get an allergic reaction or some other crap can happen. So I would apply the standards of another community, be safe, be sane and be consensual. So the short summary, for some diseases it may work or it would work. For embryos it's not really needed. And with biohacking it raises all sorts of ethics questions. Now that we are talking about ethics, MIT Technology Review had this exclusive Chinese thing. A Chinese scientist created the first CRISPR babies. What the fuck did he do? He, that's Dr. Yang Kui He. You can see on the right. He apparently had the first CRISPR babies, Lulu and Nana, that's not the real names. So the scientifically discussed names, he did one gene change in the gene TCR5, which he deactivated. Which is therefore the same, the receptor of the same name. And that receptor uses the HIV virus to infect the cell and uses the TCR5 to inject its information into the cell and make an infection. His process was like amateurish, it's not what I'm saying, but a lot of colleagues saying he hasn't published anything. But in the end of November on a conference when he had a speech there was this interesting Twitter thread by a scientist where other scientists also jumped in. They commented every single slide, commented what about this is kind of questionable and they had the results amateurish. And there were problems with consent. We don't really know if the parents, so the men was apparently HIV positive and they requested for the study and we don't really know if they knew that they are registering for a new method that was like used for the first time. But there were more problems, it was all a bit weird and the experiments were secret, but he apparently had planned a PR campaign and would say that's a bit fishy. There's even more, there's a great summary where we'll show a link, but the most fascinating thing I found was deleting TCR5 does not mean you're immune against HIV. What you're doing if you turn off TCR5 is you take away one possible gate for the HIV virus to infect the cell and many changes and deactivations will mean that you are much more susceptible to other illnesses. And there are drugs you can take that actually do the same thing that he did with CRISPR-Cas and there are a lot of methods for insemination that make sure that embryos are not HIV positive. So this is like shooting at sparrows with cannons. You can ask why, because there are so many other ways to do it. If you want to learn more, there's a great article by Ed Young in The Atlantic, he deconstructed that and there's the Twitter thread linked there. You can find it in the slides we'll publish shortly. We had the first CRISPR baby, the goal is really weird and the way is at least interesting. Yeah, that was a bit harsh. Let's go to genetics and farming. The hope here is that CRISPR-Cas will help us come up with a better plant species in a cheaper way. The way that it's more ideal allows for better plants so the entire value stream will be thus. So the main idea is we hope for a mutation that will improve a useful feature either in a farm animal or in a plant. It doesn't have to be a monogenetic feature but we hope to improve something typically very diffused and convoluted correlation. In basic research we're discovering more and understanding more how the plant's resource processing works. There will also be a talk on this topic, information biology. This talk will likely go into much more depth on this area. A second path is the genome sequencing that is also apparently following the Moore's law. So it gets faster, it gets cheaper, it gets better. Of course this is a lot of data which means we need to use big data methods to go through them and use them for something useful. And for this we need a lot of computation power. But in the end we hope to yield a process allowing us to find out which gene change causes what effect. So the end goal is having a candy store like where we can just select what we want to do and we know exactly what change we need to do to get that. We've been doing a similar process in the classic plant size where we select it and then recombinate it to improve the wild species out there. And to specially raised plant types in wild species we still find a lot of adaptations to threats to different aromas types. So something we all know is that farm grown tomatoes just don't taste like original tomatoes or wild tomatoes. Using all of the information we have gathered in basic science we can now think about it similar to how computer science works. So we can try to pick features at random or however we want and mix and match them to arrive at ideal types. Of course what exactly does ideal mean? So we like it to be tasty, we like it to be healthy, we want it to grow fast, stores well. As a society we should think about the climate change. We should think about resilience against threats like new pathogens or concrete example recently published using the wild and cherry tomato. So wild tomatoes are about pea size with only few about four in this example of deliberate changes. It was possible to increase the size greatly to about cherry tomato size. This can be quantized, the fruit weight tripled roughly and also many more blossoms meaning many more fruits on the same plant. And sometimes this is also very important economically. One example is that seeds currently is kind of like a recurring cost because you don't have to just buy it once you have to re-buy it every year. One example this is rice and it should be seeded using the wind and normally you should be able to take part of the good yield and regrow that. And recently edited rice actually allows this to do that because classically, classic gene edited rice does not allow receding and this allows the greatly increased yield to stay over many generations. While considering all of these interesting options we of course also have to look at the legislative rulings there. So for this we need to look into the past for a couple of decades. So we need to look at classic gene transfer technology. So moving genes from one plant to a different one or from one species to a different one. There are of course rules that means it's not a gene or it's not genetically modified so it's metagenesis. So by inducing a lot of change and then selecting for interesting change this is not a GMO. And in many ways gene editing is much more like metagenesis and not like a classic GMO. The EU Court of Justice ruled that gene editing is not part of the classic metagenesis. So it's not part of the selected off. But it should be in fact categorized as the same as a GMO because we don't yet know about side effects. But we already know it will be much faster and easier to employ these tools. So we need to use the precautionary principle. So we might collect our findings so far maybe a bit resigned. It's very possible to improve current species with useful mutations. But we need to find rules and figure out exactly how we're going to treat all that and how we're going to put that into regulation. But of course the big companies using GMO right now have the required attorneys to do that. OK, finally, how does CRISPR-Caswork? It's fast, precise and cheap just like Rogue One. We use CAS to mark a place to cut and if we deliver a repair template along with it we're basically done. We have seen that it makes basic research a lot easier. We can collect a lot more knowledge how biology works, how diseases work and just a lot of the basics. We can cure some diseases, some. You probably don't even have to work in the germline because there are other methods. CRISPR babies, I illustrated that with Jar Jar Binks because it's reality and you don't really know why. Thank you. We can breed organisms very directionally and many people could do that because mutagenesis would happen in the same plant so we would not have to transfer it. The problem is the legal situation is interesting. It's similar to copyright filters. It's going to hurt the little ones of course and bigger ones. I'm not going to name any names. We'll have no problems. They're just going to smile and scratch themselves on the butt thinking about their well-financed legal division. So what can it do? Well, first it can accelerate research both in basic research like the function of biology, biological systems, diseases, but also in applied science. We can see what works under certain conditions or worse. It just is faster with CRISPR-Cas. There have been a few initial clinical studies about therapies. With food plants it works very, very well in the lab. It seems that embryos have been edited. What can it not do? It can't heal anything that isn't genetic. If the initial cause is not hidden somewhere in the genome, then well, CRISPR-Cas is just CRISPR-Cas. It can't heal all genetic diseases and it can't enhance things like size, intellect, gene-doping. CRISPR-Cas is a tool, a powerful tool, but a tool. And the difference is in how we use it. And so what questions are caused by all of this? What ethics do we follow? With all CRISPR-Cas discussion, certain ethical questions that we had put on the back burner had caught up with us. They're getting more pressing because CRISPR-Cas is so fast, precise, cheap and so on. It starts with a discussion with Anna with the embryos, but it basically hits all areas we had. This discussion should have been had in many of these places already. Fear of the unknown, the reaction when we mentioned agriculture, green genetic editing technology. Not everybody on the street is a fan. I'm not saying this is out of lack of knowledge, but we have a method that can do so many things that science communication, so science itself, has to explain a lot more so people don't get scared of the unknown. I think we have to work with the unknown a bit more to make it more known. Acceptance through passivity. Maybe one thing or the other passes us by and just sort of happens and we don't really realize it. It's an open question. Will this happen maybe? CRISPR-Cas is used in so many areas. Who should decide something like this? This is me saying it. I'm a scientist. Oh, dear God, don't just let scientists decide that. That's about it with the Academy of Science. I guess I'm done. But that's how it is. Society should decide that. A mix of different groups of people. It's difficult. How fast does the law react to science? We have this European Court ruling where we went towards better safe than sorry. Interesting story. But this may not work as well with patents and monopolies by large companies. It's a thing you have to deal with, dear politics. Atomic gardening is okay, so throw a radioactive source into the environment and see what happens. But targeted mutagenesis isn't. I want to see this discussed, but put both next to each other in comparison. I can see the ethical problem even on the technical side. But how do we want to talk about it? CRISPR has a potential for democratization and decentralization. How can we use this? Why do we deign to afford locked up science? They talked this morning with topics like this. Science behind a paywall that isn't accessible to everyone. When we have things as important as this. Can we as a society even discuss this still? We thought long and hard if we put a lot of answers at the end of a talk. We would love to give you more answers, but we have these questions too. Some of them deal with science, some of them are relevant to all of us. We can't give you a lot of answers except explain to you how CRISPR-Cas works in different areas and applications. I think we've done it at least sort of properly. Thanks for listening. CRISPR-Cas is a tool and we have to talk about what we do with it. This was Anna and Catherine. My name was Anna and thank you for listening. Thanks André, Catherine and Anna. There's still a few minutes for Q&A. A few questions at the microphones. Get there fast, we don't have that much time. What we can do one or two questions. Microphone 3. I did not understand yet why CRISPR-Cas can't cause a betterment of a human enhancement of humans. I mean, if I put a gene for a growth hormone into an embryo, he could get bigger. Yes, you could do this. The problem is you have to look at how the human feels about it. And humans have pretty long generational phases. If you discuss it without ethics, you throw in a gene. You don't really know what happens if you have it double. Maybe it will be a bit stronger. So you have to look what happens with this type. It's 3 centimeters bigger or 20. But there are these problems with it. Not every protein only does this one thing. They're always connected with other proteins, other hormones, you do other things and you can't just put something in twice and the double the thing will happen. That's not how humans work. Okay, microphone number 4. Yeah, sorry. I don't see it that way. Well, my question is, people often say this will be cheap. How can I understand this? Can I do this at home? Can I play with genes? Or is that a big laboratory with 50-60 people? 50-60 people don't need like 5, a team maybe. Dollar amounts, I don't really know. But you can get this RNA in the internet. Get it in a week for 2-3 euro, the sequence. This is like what a uni workgroup with a professor could do. All I can do in China, one doctor with, I don't know, a few nurses and maybe one or two other doctors, approximately. And before it would be 50-60 people, which is a whole amount more. Microphone 1. How can I look at an organism and see where it works? You can look at viruses that attack cells. In this one paper, we had viruses that attacked a liver. How tight it was, I can't really say. You can take a virus or filter the cells, like blood cells and edit them. To say this, we did not make the liver sick. In the virus there was a CRISPR cast and we knew that the virus only targets the liver. Number 2. Thanks for the talk, for the cool talk. I saw that I do it yourself, kids, for order at home. I was asking myself, what can I do with that? Should we fear that this will be a problem? I mean, it will be very cheap. I think one of these biohackers has this and they are not really approved by the FDA. That wasn't the question. Theoretically, I don't know what these kids are specifically, but if I go into the laboratory with my old CRISPR stuff, I can do a lot of what I worked on. I could try it, but I would need... I can't really go into a bacterium with my stuff, but I could do stuff at home, theoretically. To be more precise, it depends on... if we think really villainously, like in a comic book, I want to make a death bacterium. That's not that easy. If you just have the idea and you just cut around three easy pieces, this bacterium will... can just die in the environment. It's not as easy that the question again wasn't quite clear. I could be lucky. I could just do some stupid stuff, unqualified stuff and put it out into the environment. Is that realistic or is it not? It's pretty unrealistic. Luckily, the chance with randomness is like 2 billion to zero in that area. Luckily, building a superbacteria, I feel it's like that area. You would have to know which genes to put in, so you would have to do a lot of bacteria science, which will take a few years. After that problem is you could make very virulent bacteria, but they get killed fast, so your superbacteria might be very locally limited, but randomly creating stuff is very hard. That would be a thing that nature would do for us randomly. Thanks for this final sentence. And a big applause to the three.