 For those who might be watching this on video later, you might be interested to know that during the presentation by our speakers, the student audience here in Second Life is commenting and asking questions in text that myself and the speakers can see, but I don't think is apparent on the video, so you may occasionally hear us reference some questions or remarks from the audience, and we're seeing that in the next chat. All right, well, it's a little after the hour and we have a lot to cover, so if everyone is on board, I'm going to go ahead and get started with my introduction. So welcome fellow students, welcome to the Science Circles ongoing series of panel discussions on topics of interest. Many of you, I'm sure, heard the news recently that a scientist in China edited two human embryos, twin sisters, using the gene editing tools CRISPR-Cas9. You may also be aware of the alarm that this sparked globally. Some of you may also be aware that there is an ongoing project to eliminate the mosquitoes that transmit malaria using something called a gene drive, which is a way to spread gene editing machinery throughout a population. And today we have with us two wonderful experts who will walk us through what gene editing is, specifically the CRISPR-Cas9 system, and to explore some of the applications and controversies around this mind-bending new technology. With us today is Max Chatinois, Mary Ann Clark, professor of biology at Texas Wesleyan University. He developed a genome island in Second Life for online students in genetics, and recent students have also participated in her RNA folding project sponsored by Carnegie Mellon and Stanford universities. Also with us is Steven Zudify, really, Asia. I said that right. Steven is senior research associate at Dow DuPont, who has studied biology at Tulane and University of Chicago. Steven also has experience with education in virtual worlds at virtual islands for better education. And also, Steven and Mary, please feel free to add to or amend any of your bio if I got anything wrong there. So let's get started. Mary, if you'd like, please give us some brief opening remarks to help our students get an idea of where you're coming from on this topic. And then I'll ask Steven to do the same with some opening remarks, and then we'll segue right into our slide presentation. I'll try to monitor nearby chat to curate selected student comments in voice. So, and with that, Mary, why don't you start us off with some of your opening, any opening remarks you might have. Well, first off, I'd like to disclaim expertise. If there's an expert here, it's Steven because he's actually doing Christopher work. I'm just an interested bystander. So just consider that in any, if I say anything really stupid, just consider that my bystander status. I just wanted to talk a little bit about the process of gene editing. And there are several different processes and they're not all represented here on this slide. The only thing that's on this slide is the nucleus-based editing techniques. And the nucleus is an endonuclease, meaning that it can cut the DNA within the sequence. That is, it's not from the edges. Exonuclease is cut from the edges. Endonuclease is cut across the middle. So these are all endonucleases of one kind or another. And they all also have some way of recognizing particular DNA sequence. And the first three that you see here, the zinc finger protein, the mega-nuclease and the talons, are proteins. And each of the proteins have some ability to recognize one or more bases. The talon proteins actually recognize single bases. And so a whole bunch of them are strung together to get recognition of a particular short DNA sequence. The zinc finger proteins recognize short DNA sequences, in many cases groups of three. And the mega-nucleases are looking for insertion sites to stick an intron in. This is kind of backwards from taking the introns out, if you see in messenger RNA splicing. This is actually putting the introns in to gene locus. And then finally we have CRISPR. CRISPR is different from the other three in that the recognizing entity is an RNA rather than a protein. So this is an RNA that is associated with a protein. The protein is the Cas9 endonuclease. And the RNA itself has been rather significantly engineered. And it has two important parts. There's sort of a left half and a right half. And on the left half, or the five prime end, is the recognition sequence. So that is an RNA that has been structured. It recognizes specific DNA sequence. And then the right half, or the three prime end, that RNA, you see also sort of folds up on itself. And this one is structured to recognize the enzyme Cas9. So the left end is the recognition part. And the right end is the part that attaches to the enzyme and takes the enzyme to a particular location in the DNA. And then the enzyme cuts the DNA at that point. So these are all sort of modifications of natural defense mechanism in mostly bacteria and also in archaea that keeps them from getting overrun with viruses. I'm not going to talk anymore about that. But these are modifications of natural processes. And I think I will pass it on to Stephen. Oh, did you have a question, Varagon? I did have a question. Yes, thank you. Before I just wanted to interject, I guess, before Stephen talks. So the Cas9 cuts at the location, the specific location that is attached to. But how is another gene or another DNA sequence inserted at that location? Or does that simply sort of happen naturally? I mean, maybe we'll get into that later in the talk. That is a good question. If you just cut the DNA, it's a double-stranded cut. So it cuts through both parts of the DNA. So you've got a broken piece of DNA. And there are DNA repair systems of several different kinds that will put those back together. And in the process of putting those back together, sometimes it will just introduce bases at random. Sometimes it will take the pieces that are left after the cut and use those as a pattern for inserting new bases. Or you could put in another pattern, another template DNA in there to introduce something else, something that wasn't there to begin with. Very good. There are reports of alarmingly high error rates with CRISPR. And I suspect that the error is coming with the insertion step of the process, not the cutting step of the process. The error comes with the repair. It cuts where it's supposed to cut. But what happens during the repair process is a little bit random. Okay, very good. Thank you very much. And Steven, why don't you make some opening remarks if you don't mind? And then you can segue right into your presentation also. Yeah, thanks. And glad to have the opportunity to talk about what I think is one of the most impactful things that's happened in biology for some time. I do want to start out with one quick statement, which is that even though I am an employee of Corteva AgriSciences, I'm not here as an official representative of the company and I'm not here to try and represent the interests or the products of the company. And that I am specifically involved in trying to help enhance the company's ability to perform genome engineering. And like you were just saying, some of that has to do with trying to make the way breaks and editing gets repaired more efficient. But also just understanding and trying to develop new technologies related to it. So I think this, my general comment is that now is a great time for this type of biology. That's the combination of a easily programmable nucleus. And what I mean by programmable is because RNA can be synthesized very quickly and expressed very easily in cells, we can make that sequence be whatever we want so that it targets one specific place in the genome. And the ability to deliver Cas9 and RNA into cells is also at this point highly developed in many organisms and many cell types. And that when you combine this with the complete genome sequences that we have for many organisms, this becomes this great and powerful time to do this type of thing. So, and that's why when you look at these technologies that Max was talking about, the talons and omega nucleases, these have been around for some time and they've been available, but they're costly to develop and they're also not programmable. Once you've designed a mega nucleus to cut a particular site, that's the only site it's cutting. And so this is where really the technology is taken off. And I think this is a really good time to have this panel, these types of discussions because the implications of the technology, both for medicine, agriculture, society, this is really important because of the rapid pace. If you were to go back and look at the number of publications in the databases related to CRISPR-Cas9, they're growing at an exponential rate. And these are all things related to finding new applications for it, trying to enhance the technology, or even just novel uses on it. So I think this is a great time to talk about this. So let me go on to the next slide. If nobody has any questions about that distinction between the types of editing enzymes, then I think it's good to move on. Yeah, very good. I appreciate those opening remarks. Let's do it. Let's get into our presentation. So the next slide is giving a little more close-up view of what CRISPR-Cas9 does, and the RNA is represented by the yellow. Again, programmable, those bases that are able to interact with the DNA that's in blue are something that anybody can kind of design the sequence you want based on a known genome sequence. And the protein, which is kind of represented by the orange, can come in and cut both strands of that target DNA, and Cas9 does it in a very particular sequence compared to, again, where that yellow is being represented. And the thing is that this is all it does, and as you're getting into the discussion, all the enzyme does is it breaks. So really, what we call the field genome editing or engineering, the enzyme itself is really just genome breaking. And so the vast majority of where our initial uses are coming from has to do with understanding applications where we're just messing something up. The other thing is that you can, so in terms of just like the very specific repair process, there's something known as non-homologous end-joining. And when a cell has one of these types of double-strand breaks, this repair process called non-homologous end-joining goes in and tries to put the DNA back together. And sometimes it'll just put the same sequence back together, but what then that gives, that gives the Cas9 an opportunity to recut the sequence. And so the interplay that happens is the sequence will keep getting cut until the sequence changes in a way that's no longer recognized by the RNA and the enzyme. And the way NHGJ usually does the mistake-prone repair process is a small alteration, maybe the addition of a nucleotide, the removal of a couple nucleotides, and that is what you're left with. One of the interesting results recently in the literature is that you actually, based on the sequence you're trying to target, you can have a pretty good predictive ability to know exactly how it will mutate based on the sequence contact with Cas9 breaks. So, but that's what we're limited to. Now, let me just go to the next slide. Stephen, do you mind if I interrupt here for a second? Just to, if you don't mind, just to follow up on a question. So, what you just said, it sounds like the CRISPR-Cas9 assembly sort of is just kind of lingering around, and it's going to cut whenever it identifies a sequence that it can bind to. So, of course, if it just rebuilds the same sequence, it's going to cut it again. But does the CRISPR-Cas9 assembly ever get sort of cleared from the body, sort of degraded somehow, so it goes away? Or is it kind of always there? Yeah, so most delivery mechanisms are to create transient expression of the protein and the RNA. And RNA is not a very long-lived molecule in the cell. And so, actually, one of the, yeah, I can see that very quickly by RNA nucleases. There's actually a lot of effort in the field. If you want to go to any sort of RNA company website, one of the things that will have their splash page are ways they modify RNA to make it more stable so that, you know, doesn't just get randomly degraded. So there's usually this very limited lifetime. Now, there are other ways of doing the technology where you integrate the genes that express Cas9 and integrate the genes that express the RNA. But I think the majority of use cases we'll talk about have to do with either one performing some sort of transient expression of the protein of the genes, or even just directly injecting the protein and the RNA as what's called an RNP, a ribonucleoprotein, sorry, ribonucleoprotein particle. And then that, yeah, the RNA gets degraded relatively quickly within maybe 24 hours. And the protein itself also, over time, gets old, misfolds, and gets degraded by the cell as well. Very good. Thank you. All right, please continue. So the consequences of Cas9. And so this slide is just trying to talk about the different outcomes that one can try and have from this. And so we see the Cas9 RNA interacting with DNA and making a break. And that's what you see in that after the first gray arrow. And then the gray arrow to the left is in situations where you're mutating the sequence in some way. But what you have on the right is attempts, and we'll talk about this as a separate, like as Mary, sorry, as Max was saying, there are these templated ways of repairing a break. And what you can do at the same time is also inject a DNA template that has sequences similar to the target, but then also have alterations. And you hope, and these are typically infrequent events where you hope that that red sequence, your template gets incorporated instead. And so what's important to recognize right now with the state of technology is that we can do this templated insertions in situations where we have a population of cells we're altering, and then we can look through the cells and pick the ones that we want. Whereas when it comes to, say, one or two single cells being edited, say, like in a human embryo or one or two important stem cells, then that's not a great technology at the moment. And so right now, in terms of this year, we'll talk more about the things that have to do with, like, altering the gene. Because I think, so as a way of doing this by analogy, you know, if you think about, if you have a cloggy pipe in your house, there are two ways that you can fix this cloggy pipe. And on the right, if you get a template, right, you get a whole fresh new pipe, you take out the old one, you replace it with a perfectly functioning working one, then you're in great shape. By analogy, the left-hand pathway of just trying to break stuff of, you can imagine leaving that pipe intact, but what you try and do is go in and break up the clog, or you break up the amount of debris that's going into the clog, or even maybe make some sort of, you know, device that goes through and removes the clog temporarily. And that's kind of what we're doing right now is we're not necessarily fixing the system, but we're trying to find a way to break it gently in a way that benefits the cell or the organism. Oh, let me just comment that not homologous enjoying, which you have here is more or less the way your antibody genes get produced during development. Yeah, so it's an important repair process, and these things are adapted in various different ways to be used to the cell, but that's an inherent repair process that we just take advantage of. Right now, it's incredibly difficult to alter the way cells do the repair mechanism, and that's why templated repairs are so difficult. So, all right, should we talk about applications or any questions maybe about the technology before we go on? I think we're okay. You don't really see too many comments from nearby chat. Let's go ahead and move on to the next slide. So, here is a very interesting use case that had a new splashy result this year. And so what I'm going to talk about is the muscular dystrophy, and the protein that is altered in muscular dystrophy is known as dystrophin. And its general function in the cell is to basically be an anchor between the outer membrane of the cell by connecting to a membrane bound protein. To the internal skeletal structure of the cell. And so it plays this very important structural mechanical role in the cells. But it's also huge, and I don't know why it's so huge, but it is, I believe in terms of the amount of genome space it takes up, the largest gene in the human genome. And we are debating about this. We also are pretty sure it's the largest protein as well. Now what happens in that, in many cases of people afflicted with muscular dystrophy, there's a small alteration in the coding of the protein such that it gets truncated. And so this is something known as a premature stop codon or a frame shift mutation that occurs. Now, and then that basically you don't have strong anchoring within muscle cells. And so you have basically very weak skeletal muscles as well as weak smooth muscle in order to perform all those really vital functions. Now the organism can still develop, but again with a very weak muscle structure that again is highly lethal at a young age. So the strategy, and these are strategies that actually have been pursued in other, for decades in this model of disease, where you try and provide some at least basically altered way of expressing the protein even though it's not the normal full protein. And what we have here, one quick reminder for everyone is that all of the proteins that get expressed are basically small segments of exons that encode that protein that are interspersed with sequences known as introns. And so it's the splicing together of all the exons into one RNA sequence that leads to the actual coding of the protein. And what you can do in this case and what the kind of history of research has been is to try and find a way to take out one of those chunks, one of those exons and then re-establish the vast majority of the protein being expressed even though you might have an altered or even missing small middle part. And so that's what the strategy is showing down here is that if you have an exon and then you have a mutated exon that's pink, what you can do is just get rid of that pink one and then re-establish from exon say 47 to exon 52 a reading frame that works to express the protein even though you're missing some middle parts. And so Scissor G asked me to define an exon and I'll just repeat that that's the DNA sequence that specifies the parts of the protein that get made. Yeah, if you don't mind, I'd like to jump in here too because I think perhaps many people don't appreciate that a gene that is transcribed into a protein, the gene is not just a continuous sequence of base pairs that code for amino acids that become a protein. What's weird about the system is that the protein is coded for in the DNA by sort of sections of DNA that are interrupted by introns that are just kind of inserted in there. And so there's a kind of complicated process to sort of transcribe the protein without the intron portions. You transcribe it with the introns and then take the introns out. Thank you, yes. So the process is kind of counter-intuitively more complicated than you might expect because nature uses what's worked. It's not intelligently designed, it's just what we have and we're making it work. Sorry for the editorial comment there. And I'll shut up and let Steven and... Oh, yeah, yeah, no worries. And I think one thing to keep in mind that's powerful about these abilities of splicings, that you can have splicing variations that lead to new proteins or variations that are very useful for evolution. So the result that came out this year, and this was something by the Olsen Lab, is there's this colony of dogs that have Duchenne muscular dystrophy. They're a great model organism. This is I think in Cambridge, Britain, England. And so what I'm showing here, this is staining the dystrophin protein in muscle tissue. And what you have on the left, this nice little green honeycomb looking structure, is a normal diaphragm tissue mass with dystrophin nicely arranged around the cell membranes. What's in the middle is a dog with Duchenne muscular dystrophy, and you don't see that structure. And if you would actually look at the phenotypes of these dogs, they don't walk very well. They can't jump. They are highly disabled as a good model example of the disease. And what's on the right-hand panel is the muscle tissue from a dog that had one single injection of viruses... Again, engineered viruses that express Cas9 and the RNA delivered to the bloodstream and then allowed to go into cells and try and do this editing fixing process. Again, similar to what I was talking about. This is not full dystrophin protein. This is the full length of it with some maybe missing parts in the middle. And the staining pattern is re-established. And in fact, the phenotypes of the dogs are to be essentially normal as long as the injections are given at a relatively young age. And so this is an example of trying to find a way to kind of alter DNA sequence that actually fixes, or at least largely fixes the expression of protein to then basically restore normal function in dogs. This work has been done in mice. This work has been done in human tissue culture lines. And so it seems to be relatively reliable and relatively robust in terms of the medicinal application. So this is amazing. And again, there is a video that I think is behind a paywall and I've been unable to re-establish it, but also presented these at the Cold Spring Harbor CRISPR conference back in August. And I was there. And it's just amazing watching this. And hopefully there'll be more media related to this in the near future. Yeah, so I think, you know, there's this question from Bob Crescendo that one of the hurdles of any sort of gene therapy strategy is the ability to deliver your agents into cells, into enough cells and with enough power to do what editing you want them to do. And that in many cases limits the applications we have. But muscle tissue is something that's very good at basically uptaking foreign DNA. So you can actually just do DNA into bloodstreams and get it to work or intramuscular injection. Other cases, you know, we have to design viruses or design ways to deliver things to particular tissues. Again, CB Axel did the dog get better. Yeah, the ones that were treated with the CRISPR were basically normal dogs up into the age that they were examined. So I think this was something they've done it this year, so they're still less than two years old, so they can monitor their progress and I'm sure there'll be updates in the future. Okay, so let's go on to the next one, because I think that kind of transitions into the next thing we're talking about. That I think one of the other interesting and exciting developments in human gene therapy models with CRISPR is something coming out of, let me grab the link, another lab where they have found that if you take T cells out of a person, and again these T cells are things that float, they're not anchored, they're going around monitoring for invaders in your bloodstream so you can grow them in culture relatively easily as well, that they did electroporation where they used an electric field to inject the Cas9 protein and RNA, as well as a DNA template. Again, now we're talking about templated, trying to get a template to be used as a repair into T cells. And they got this to work more efficiently than has ever been done before, such that the efficiency of knocking in the gene was about one-third to two-fifths of a time. And so this is pretty amazing. And then, of course with those T cells, you can grow them, you can monitor which ones have the alteration you do and don't want, and then re-inject them back into a person. And so the particular modification they were working with in this case is injecting in the specific antigen for a tumor, I believe it was a type of melanoma, and basically getting that expressed as part of the T cell receptors. These are antibody-like molecules that help recognize and destroy invaders or aberrant tissue like cancer. And so they found that when they altered these T cells and inject them into mice with a model of the tumor, they actually got tumor suppression. And so this is combining, I think, one of the other amazingly exciting fields right now, which is cancer immunotherapy, that we can use CRISPR-Cas technology to take a person's cells, take them out of their body, alter them in a way that can attack a cancer that's inside their own body. And because we're using their own T cells, there won't be any immune rejection of the therapy. And so, again, that's the promise of this. And the ability to actually put in whole chunks of templated DNA is a key part that's a new development, I think, in the field to at least work this efficiently. Any comments from Max or the audience? I noticed that the slide says that it clears cancer cells and rates comparable to those observed for T cells engineered via retroviral delivery. Can you kind of help us put that in context a little bit? Okay, yeah. So a retrovirus, in this case, what you're trying to do is get a virus to go into the cell you're altering and actually integrate it into the genome. And then what that retrovirus is doing is expressing a whole chunk of pre-engineered DNA, like the T cell receptor with the antigen, the antibody you essentially want, into the cells. And so that's a very efficient way to put in a trans-gene and to express, again, a whole genome construct that includes the promoter, that includes the sequences that you're trying to get expressed. And so, again, that's a great way of doing genome engineering in a lab, but the problem with retroviral therapies is that they can, in certain places, they can be very dangerous for an organism because those altered T cells with the retrovirus could actually become cancerous on the run. So that's why CRISPR is better than retroviral therapy. Yes, Marianne. I think one of the interesting things about this somatic gene therapy is that you don't have to fix all the broken cells. You just have to fix some of them, enough of them to, you know, produce whatever it is that needs to be produced normally. Very good, very good. Even if you don't fix all of it, if you fix enough of it, it seems to work. Yeah, very good point, thank you. It's a little bit like having both normal and abnormal proteins if you've got, you know, one normal gene and one abnormal gene and you're going to be making both proteins, but the one that works is the normal gene. And I'm sorry, I just... It's one that's not working, that's the problem. And I'm sorry to blink on this, but this is from the Morrison Lab, at the University of California, San Francisco. And I think in collaboration with the, what's called the Chan Zuckerberg Biohub, also in San Francisco. So, make sure you get the credit properly due. Thank you very much. Okay, let's move right along. I want to keep my eye on the time, so... Oh, here we go, patents. So, Steven, I might let you talk maybe specifically about this particular patent fight I might have injected. I am a patent attorney, but I have not really followed the patent cases that closely. I thought maybe just to make... I might want to give a little background to our students in general on some basic patenting principles. Patents are actually protected in the U.S. Constitution. Patents and copyrights are specifically called out in the... Patents are for inventions that are new, useful, and not obvious. And in patent law, obviousness has a very specific meaning. Patent applications can be rejected. There are basically two primary bases upon patent applications. One is that the invention was already invented by somebody else. That's called anticipation. So, if a single reference describes your invention, then that reference is said to anticipate your invention from the basis of... The other basis that a patent application would be... Obviousness. Obviousness has a very specific meaning. What that means is that the examiner, he can't find... He or she can't find a single reference that anticipates your invention, but he can find the elements of your invention by combining two or more references. And when that happens, he says, it would have been obvious to combine the features of these other inventions to make your invention. Now, obviousness rejections are difficult to overcome. Back in the day, in order to support an obviousness rejection, the examiner had to cite something in the public domain that suggested that the combination of references was sort of the logical thing to do, like some paper that said, well, gee, maybe if somebody combined these things, this might work or something. However, that rule where you had to provide some basis from the public domain suggesting that the obvious was struck down by the Supreme Court about 10 years ago, primarily on the basis of pressure from the software industry, which felt that the Patent Office was granting too many obvious software patents. So now it's very difficult to overcome an obviousness rejection unless you can amend your patent application to recite an additional element that is not anywhere in the prior art. The prior art being any published document that was in the public domain prior to the filing date of your... So that's a little bit by way of background to help you understand when we talk about the fact that the competing patents for CRISPR technology with respect to obviousness, that's the context that we're talking about it in. So with that in mind, Steven, if you'd like to go ahead and walk us through your slide, that would be great. Yeah, and I really don't want to spend too much time on this overall in that just a new development occurred this year in terms of how this is probably going to be resolved. But to give a brief timeline on why this is important in science and in terms of the adoption of the technology, is that really there was a lot of foundational work in terms of understanding CRISPR domains done for... Actually, since the mid-1990s, I think. But it was Jennifer Doudna. And this is a name people have probably heard the most about, who along with Emmanuel Charpentier published the ability to engineer Cas9 to target sequences and cut it. And around the same time, Virgus Sisgnis at Vilnius University published similar results. And this is working in a test tube. But... And they filed patents, of course. But it was in early 2013 where Feng Zhang of the Broad Institute of MIT and Harvard published using CRISPR-Cas9 in eukaryotic cells. And they, of course, also filed patents. Now, what they did was they paid extra money to get it fast tracked within the patent office. And this is one of the competing things that happens in the patent office. So if you have money, you can get your stuff reviewed faster. And they got their claims granted. And, of course, UC Berkeley, which was basically the patent filer for Jennifer Doudna, sued and said, hey, you can't get all that. That's what you're doing is obvious compared to what we did in the test tube. And, again, through rounds of litigation, the most recent ruling by the Patent Appeals Board in February of this year said, you know what, it's not... To some degree, it's obvious what to do, that you, of course, would want to try and give this work in eukaryotic cells. But the ability and the technology to make it work well and demonstrate it, how you do that, first of all, it's not guaranteed. And there's a lot of work that only an expert would be able to do to get it to work. And so I think that's where the claims currently reside and how it was resolved. Again, there's still another round of appeals to the U.S. Supreme District Court, but most people don't think UC Berkeley will prevail. It's possible... So right now, as it stands, if you were going to commercialize anything that had to do with being edited by CRISPR-Cas9, you need a license from Brode. And that's basically the way it stands right now. It's possible that the Dow and the Group will also have their claims granted and that anybody wanting to do any editing and commercialize it in eukaryotic cells will need a license from both UC Berkeley as well as Brode. Now, just to briefly touch on the Brode license, they are and have said they're committed to making sure research into CRISPR-Cas9 and the development of the technology is widely and freely available. Again, there are lots of people who are going to patent specific uses of Cas9 or their alterations to the technology or ways of using it that Brode will have an umbrella patent over. But, of course, they do have very specific restrictive licenses for commercial entities. And so, without going to the details of this, I just want to make sure everyone's aware that there is this kind of gatekeeper pathway for anybody trying to commercialize products related to Cas9 editing. And that, of course, they actually do have a bit of a gatekeeper that if there's a really good idea of a gene to try and alter and edit for medicinal purposes, they actually get first crack at it with a related commercial entity. So I think I just want to leave it at that so people are aware of it. That's kind of the most latest development in it. Although, again, most people in the scientific community really do credit Sysnes, Vilnius University, and UC Berkeley has really been the inventors and they'll probably be the ones who get the majority of any sort of academic or award accolades. Fantastic. That's really interesting. From what I've read, it does look like the commercialization of CRISPR will probably move forward. That is that Berkeley and the broad people will probably form some kind of a patent pool or some kind of cross-licensing arrangement so that people who want to use this technology can sort of obtain a single unitary license from both entities. I also would like to point out that one of the reasons the ability to get CRISPR to work in eukaryotic cells is interesting is because, as we mentioned earlier, CRISPR is a system that evolved in bacteria which are non-eukaryotic. They're prokaryotic. They don't have a nucleus. So it's a little bit surprising that a system developed in a non-nucleated cell would be able to work inside the nucleotide eukaryotic cell. But what we've discovered, in fact, is that it appears that the CRISPR system is so ancient, evolved so early in the development of life that it is essentially universally applicable to all life forms. So even eukaryotic life forms, that's just, I think, kind of philosophically quite interesting. I'm just going to comment that the genetic code is more or less universal. So if it's going to work in prokaryotes, it's probably going to work in eukaryotes, too. Very good. Thank you. So this is actually a good transition point where even though you might have to pay money to brode to commercialize something, the value of your product could be so big that, of course, it's worth it to pay the licensing and royalty fees to some other entity because it's such an amazing technology. And this is what agriculture as an industry is facing right now is that, of course, we have to, you know, pay licensees, royalty fees for a number of things to put a new crop out in the market. But of course, one of the other big costs to commercializing an agricultural product does have to do with regulations in different countries. And so, again, I don't want to spend too much time on this one, so I just want people to be aware that there's new stuff that's occurred this year in this big society debate that the USDA said we are not going to regulate CRISPR engineered products if the type of genetic alteration is something that could have been a result of traditional breeding methods. And so if you're going in and making point mutations or altering the expression of a gene that's known to behave a certain way in a plant when it's mutated and you basically recreate that type of mutation or that type of alteration, it doesn't undergo any sort of special regulatory review or processes. And so that's great. Now, the European Union has actually taken the opposite stance, which is that even if you're altering any sort of mutational activity that you're engineering into a plant for a desired result, that will be regulated as if it were a genetically modified organism. Now, let me point out that the term genetically modified organism has a very specific meaning in the regulatory documents, meaning the insertion of foreign DNA. And so the U.S. says CRISPR editing or CRISPR mutations are not the same as the GMO. Whereas the EU is saying they are the same as the GMO. And regulatory hurdles add time as well as cost to anything that you want to basically have grown or be a commercial product in the specified areas. And so the European Union is something that people were actually not expecting. People thought they were going to rule the favor of being similar to the U.S. And so this is something that, you know, there's still a larger societal context to how we view altering organisms that is, you know, debated and very different depending on your country and culture. Well, okay, yeah. Yeah, that's a little bit, I mean, does that suggest that in Europe, even if you were to, it's not sure what I'm trying to say here, but what if you were to replace a defective gene with a working endogenous gene from the same individual or from the same species? I mean, would that still fall outside the European Union guidelines? There would still be a GMO? No, well, yeah, so that's, you're talking about something that's cisgenic, but it's a recombined type of sequence. So that's something that is still regulated, considered a genetically modified organism in terms of rearranging and redoing how the DNA is done. Okay. So let's move on, because the most important part of our topic, we need to leave time for it. I'll mention that in terms of the agricultural business, there are several companies, none of the big ones that right now in the field are releasing for commercial development and processing, or those last bits of field trials with some interesting CRISPR-edited products coming to the market. So look for that, maybe we can have a different talk about it sometime, but these are things that alter our food supply and that are coming out as a result of CRISPR technology. Okay. Last slide. Everyone's probably been waiting for this. Yeah, so, I like that first name. Dr. He, H.E. from China, basically released a YouTube video saying we have altered the genome of two people that are now born. And let me just mention the science, which is the targeted gene in this case is something known as CCR5, and CCR5 is a protein that exists on the surface of T cells, and it is a co-receptor molecule that the HIV uses to get inside cells to do its dangerous stuff. It's a retrovirus that integrates into DNA, but it can't do that unless it gets inside the cell. And its mechanism of entry is this protein. And so, and I gave a CRISPR talk earlier in the year, and what is known and why this is known to be a good target for this is there are individual humans that exist in the world that have this mutated already and are highly resistant to getting infected by HIV. And at the same time, there are people who have one good copy of the gene and one bad copy of the gene that actually have, they can still get infected by HIV, but the progress of the disease is significantly slowed down. At the same time, there have not been any major defects noted in individuals homozygous for this mutation in terms of any sort of loss of fitness or fecundity or any sort of clear phenotype. It seems to be something that you can delete from an individual with largely no consequence in terms of the gene itself. And so, the reason why these particular individuals were chosen to have, or what should I say, the parents were sought out and said, hey, we can offer this to you. And the reason why it was offered to them, at least in the case of these particular parents, is the father has HIV. And so, there is a significant risk of the children developing HIV through any sort of accidental blood-to-blood contact with the parent. Again, I don't actually know what the actual risk of that is. I don't know what surveys have been done. But the attempt here is to say, these are children that can never get HIV and never have to deal with either the disease or the stigma of having the disease. Okay, yeah, so actually, I should mention that. So, since Reggie asked, has it been verified, is it just a hoax? Fair enough that he has not released all the data in a peer-reviewed publication as you'd normally expect for this type of information. My understanding is he may have somehow been prompted to try and release this information earlier than he'd really meant to. Now, there are scientists who have reviewed what he has said he's done, right? There's scientists out there who have criticized because they've seen slides or parts of the data of what he says he's done. So, I think it's fairly likely that this actually did occur. But the actual verification by other individuals, that is to say, take these children, these individuals' DNA, and actually analyze it by an independent lab, has not been done. So, that is a good point to mention right now. He has passed it in that process, so somebody's looked at it. Oh, that's interesting. Teal asks, what about the children of these girls are edited genes passed on? Yes, and that's the goal of this. Now, again, one thing to keep in mind is that in theory, you're trying to make these individuals homozygous recessive for this gene, which means that their children can at most only inherit one bad copy of the gene. They would have to mate with somebody else who also has bad copies of the gene for that to be, you know, a completely protected offspring. Or somebody who's got the other allele, the one that's already out there. Yeah, and that's maybe I should point that out too, that when we look at the frequency of mutations in this gene, if you look in Europe, about one-third of people have mutations in this gene, and that's because CCR5 also provides protection to bubonic plague. And so there's actually like the reason the black plague killed lots of people, but then subsequent epidemics killed fewer and fewer people is because a natural genetic resistance was being selected for within European populations. And so that's an interesting great story, but I think to get back to the main point, we are talking about the germline alteration that changes the makeup of the human genetic gene pool that is permanent and something passed down from generation to generation that then undergoes selective processes. Or again, if some individual wanted to to go through more rounds of genome editing. So since we have a little bit of limited time, I'm going to jump in here and I want to, since I have experts here to ask this question of, so my stance is that in maybe three or four generations, designer babies are going to be commonplace. I think that and because CRISPR is so cheap that it won't even be limited to the rich, that in fact it will be extremely widespread. And I wouldn't be surprised if in, I don't know how many generations, three, four, five, six generations, that essentially the human species is just inevitably going to be transformed by this technology. So what do you think of my proposal? Well, let me say one thing. I think there is a certain amount of inevitability with this because it is so powerful and potentially so useful. But I think now would be a time for Max to put up her second slide to talk about, you know, how other experts in ethics and society do view the technology. Because so in 2015, there was a report of some genome editing of embryos that were not subsequently implanted. However, it did prompt a large number of people of experts in the field and ethicists to sit down and discuss this and say, what sort of guidelines can we come up with? So National Academy of Sciences, of course, was one of those groups. Yeah, and the slide's coming up now. Thanks, Max. And I'll let you kind of take that next part of the conversation. These are those guidelines. And I think that 2015 was an important year because I think science had CRISPR on the cover in 2015 as the, you know, advance of the year, the breakthrough. So it's been recognized now for several years that this is a technique that's going places. And because it is so powerful and it is in some ways simple, I mean, messing with human embryos is not that simple. So CRISPR, getting cells altered with CRISPR may be relatively simple, but dealing with embryos requires a lot of manipulation. But nevertheless, it's obviously possible. It's apparently been done. Stephen mentioned that some work has been done with human embryos that were not implanted in beta thalassemia, I think, and they did manage to correct at least some of the cells for that particular disorder. But when people started messing with human embryos, it became obvious that there was going to need to be discussion about, although the Chinese babies sort of exploded upon seeing before that discussion had been totally pursued to its resolution point. But these are just some of the principles that were suggested by the National Academies for thinking about in looking at manipulation of human embryos. One of them is to promote well-being, which means basically you're not sick. It doesn't mean that you're, you know, beautiful and athletic and you can sing really well and you're very artistic. It doesn't mean designer babies. It means promoting well-being, health. Yes. That's very interesting. And I think, you know, I mentioned earlier at the beginning of the talk of the project to use gene drives in which the entire CRISPR editing tool is inserted into the DNA so that it can reproduce itself and spread throughout a population. There's a project to use that to eradicate the mosquitoes that cause malaria. And I think that process, the malaria project, is actually making an effort to follow these guidelines unlike the designer babies. You know, when you're working with non-human organisms, it's a little less touchy. Although I'm a little nervous when I hear about eradicating an entire species because we don't really know what else those mosquitoes might be doing. They might be doing things that are important to the well-being, not just of us, but of other organisms. I think it'd be fine to spread the, you know, resistance to the parasite. That'd be great. So, yeah, you do kind of... Kill them off the whole insect species? Right. You kind of invoke chaos theory where the butterfly effect where a change like that could have unpredictable consequences. Let me touch on a couple of things from the students. Aurora mentioned that even though CRISPR is cheap, it's patented and therefore the commercialization of it could be quite expensive, which sort of does kind of...which is a good point and sort of does color my view that it could be... that the designer babies could be widespread. And then there was another one... Oh, another student, I think it was...Bong was worried about using...that our diversity would suffer as we increasingly design ourselves or edit ourselves. My kind of view about that is that, I mean, in a sort of totally dystopian situation, that if we go forward, diversity slowly becomes less relevant because we can simply engineer any adaptation we want. We don't have to find the adaptation in nature. We can just engineer it. But you guys may have a different view. So, what's your kind of view on how this might affect human development? I don't think it's going to be accessible enough to most people to affect diversity a lot. I mean, yes, I would worry about affecting diversity. If everybody decided that they... I can't think of any, you know, stars or something that you'd want your kids to look like. But I think that that might not be as big a problem as the fairness issue. It's the accessibility issue. Yeah, I agree that we don't necessarily want to promote heavily a technology that can largely yet again increase the differences between rich and poor people. But in terms of diversity, I mean, if we think about the 100 or 500 year view of this and the pressures that the human species may have, well, if somehow AIDS would become a hugely prevalent problem, then it would be very good to have low diversity in terms of the CR5 gene and to again have found a way to make the technology more widespread for people and that that would be a good thing. I think getting rid of genetic disease is not a bad idea. I don't know if we're going to get rid of it, but getting rid of some of the, I guess, the high profile genetic conditions that we understand well enough that we might be able to do. I think it's very useful by case dependent. But I do think we need to think about this in a 500 year review of what are we trying to establish now for what's better for all of humanity for the indefinite future. I'd also like to make a plug for genetic diseases that not just the high profile ones, but I actually think one of the key uses of gene editing for genetic diseases is for rare diseases, for orphan diseases, for which, you know, that aren't profitable for drug companies or the medical industry. Two cures for, because there's just no money in it. That's a nice point. I have a personal, it's taken this because my brother has an extremely rare form of CMML, CMML2, which is caused by a mutation that, you know, just came out of the blue, but it's very rare and there's basically no cure for it. So, and it seems to me it is the perfect kind of rare illness that you feel for cures. Catch it before it gets out of control, you mean? Yeah, yeah, yeah, or just edit it out. That's an interesting idea. You get a bad new gene, cut it off with a root. Yes, that's right. Yeah, well, and this is, so, you know, Aurora kind of mentions this in the chat, that, you know, the father, the parents made the choice to do germline editing as compared to a number of widely available ways to deal with HIV somatically. Like you can either think about somatic gene therapy, or again, there are cocktail drugs that help suppress the disease. But I, you know, my perspective on this is that we can talk about fixes or we can talk about band-aids and that germline editing is really the one way to take, you know, just to have people like your brother never suffer in the first place and fix problems. And one thing we also don't want to do is set up, I mean, the cost to society of saying, well, every individual who gets born with this disease will treat them with somatic cell therapy. Then, you know, that's a larger cost to society, probably a small number of companies that are making lots of money off of doing that same thing over and over again, as compared to trying to find ways to, you know, do germline editing, which in terms of cost to society for these would overall be cheaper, and also probably, at least compared to the risks of the technology as we understand it, have low downsides for each individual in terms of their, you know, what they're suffering through in their lifetimes. Yeah. Let's see, I think we still have, let's kind of linger here a little longer. I think we can kind of extend our time a little bit. Yeah, let me just interject. Let me go ahead and throw up the last slide that I had prepared ahead of time. Fantastic. As a context for conversation. Fantastic. A few weeks after the result, the children were announced, there's been a lot of discussion, and Ed Young kind of put together, I think, the best that I saw as critical as possible conversation article in The Atlantic. And so he gave about 15 different points of what really bothers him about the announcements and how it was done. And a lot of his views do have to do with kind of the process that he went through in terms of ethics and transparency. And again, some criticism of the biology and the technology in and of itself where he again, did ask some experts to weigh in. And there are, if you go on, you know, social media, there are a lot of scientists who are really extremely aghast at the fact that there was any German editing done at all. And I think it's fair to say, you know, he did violate this 2015 larger agreement to not do this. But on the other hand, it's now been three years later. And, you know, there are great targets. Well, he's missing now, right? There are wonderful great targets. We know that we can start helping people right away with this type of German line editing. And there really has not been a lot of conversation about it. So I think, again, one thing I do want to say is that, you know, they're made to, in my mind, there's a level at which we need to say there are things standing in the way of a good thing. And there need to be people who are ready to try and break through kind of a stalled conversation. So again, these are points to talk about. Let's get some discussion going on. And I'll let Bergen get it. Hey, so thank you very much, Stephen. That's great. And I'm a big fan of Ed Yong. So I think I appreciate that's a nice touch. There's a little bit of discussion going on in local chat about, you know, how widespread CRISPR might get about whether medicine just becomes increasingly more expensive or, in fact, whether less expensive over. I'm not quite sure how to evaluate. I just will point out that patents have a limited lifespan. Part of the bargain for getting a patent monopoly is that, one, you have to teach the invention to the world so that other people can learn and improve on it. My patent documents have very detailed explanations of what the patent does and how it works and how to use it. And then finally, they have a limited lifespan. They do eventually all enter the public domain. So eventually, the broad principle, broad CRISPR patents will eventually, those patents will expire. That will bring depression. Theoretically. Anybody who already owns intellectual property will put that much more money into trying to extend their monopoly and licensing agreements for as long as they can. So one thing to keep in mind is that, like you're correct, the patent system is designed to work that way. But there's a lot of money and effort to go into, you know, trying to maintain a monopoly status over technology as far as long as possible. Well, yes. And I will temper my remarks by pointing out that the pharmaceutical industry in particular has gained the patent system ingeniously to maintain high prices for medicines and extend their patent life. One way they do this, for example, is they initially get a patent on the molecule, right? And so then that patent lasts 17 years or however long. Then they get a patent on a, like a powdered formulation, like some formulation of the cell form or something. And then 15 years later, they'll get a patent on an injectable form of the drug or something like that. So they keep finding ways to game the system and their patent. So, right. So that does go on. And it is certainly an issue that will complicate the availability of gene editing therapies for sure. There was a question that we had in the local chat asking if we could use CRISPR to alternate DNA repair, I guess, to make CRISPR work better. And yeah, there are ways you can do that, but you really don't want to, like, the most you would want to do is to alter the DNA processes during that short period of time that you're trying to do to get CRISPR to do stuff. And so you don't want to long-term alter DNA repair processes because right now- It could have serious consequences. Yeah, it's kind of at this right balance of what it is and is not supposed to do at any given time because DNA repair done the wrong way in too much is actually really bad for a cell. It could lead to, like, chromosome alterations breaking up and recombining DNA in much worse ways. So I think that was a very good question. But just keep in mind that, you know, certain alterations you want to just avoid being permanent. Actually, maybe, okay. But one other application of CRISPR-Cas9 is instead of breaking DNA, you can add things that are known as transcriptional activators or transcriptional repressors to turn genes on and off. And so there are ways where you can transiently alter the expression of a gene. Now, that's usually not as valuable in a medicinal therapy type of way, but there are examples where one can imagine that would actually be quite helpful and that if you can make that a cascade, that once you turn some genes on and they persistently stay on, that there are, you know, targets you can think about there that are not, like, genetic alterations. Steven, could you talk a little bit more about gene drives too, maybe tell our students a little bit more about how gene drives work and what they do? Oh, yeah, yeah. So this is really cool. What the idea here is, let's say you alter the genomes of a bunch of male flies so that when they mate, they pass on a gene where then... That alters the babies. Yeah, alters the babies. The problem there is that that's only going to get passed on to a bunch of females at one period of time, then the males go away. And what you really, and that the inheritance of a gene is only half the time in general. That even if you made a male fly homozygous for some gene as it gets passed on, the next generation will only pass it on half the time. So the idea of the gene drive is that in the offspring, you pass on this heterozygous, so you pass on this gene that's heterozygous in the offspring. But then the CRISPR-Cas9 targets the double-strand break to the other copy of that gene. And then it's repaired using your inserted version. Yeah, so then basically every generation has this homozygous for this gene drive. And that gets passed on to the males who are still fertile. And so now you have these homozygous males that are passing on to the next generation. And 100% of the time, they mate and have offspring that are no longer fertile in terms of females. But the males are still fertile, and then the next round of breeding means another round of females have offspring that are not female. Now again, one thing to keep in mind about mosquitoes, which is really important as a model organism for this, is it's only the females that actually bite and pass on malaria. So by keeping males around that are not harming anyone and allowing them to basically mess up the fertility of a population, you can eventually make the population crack. Because then in those final couple generations, there are only males around with a bunch of infertile females and boom, you're done. Thank you. Yeah, thank you. I kind of thought the gene drive that sort of, there was a mechanism where normally CRISPR, you've got the RNA that's going to bind to the site that you want to cut and then the Cas9 is going to cut it. But the gene drive itself is sort of that system, the system that codes for the RNA and the Cas9. The DNA that codes for that whole system is inserted into the organism that you want to edit. And therefore, then that's how the editing system is propagated from generation to generation. That kind of how it works or am I oversimplifying it? No, that's correct. What you have is you're actually inserting the Cas9 as a trans gene. This is one reason why no one would say do this in humans. It's actually a trans gene that's persistent and it's just altering the rules of inheritance. Instead of having an allele stay the same frequency from generation to generation, you're basically making it 100% of the time that goes into the offspring. You actually increase the frequency in the allele. And the reason this works, and let's come back to that, is we as diploid organisms, we get one set of chromosomes from our mother, one set of chromosomes from our father. And in our cells, if we have DNA damage to one of those two copies, it then copies off of the other chromosome. And so if there is some difference in between those chromosomes, again, if the normal chromosome is broken, then it will eventually try and use the other chromosome, this heterologous sequence chromosome as the repair template, and thus that copies the gene. This is also something in cancer biology known as loss of heterozygosity. But that's how it works. Okay. Mary Ann, did you want to remark on that also? Uh, no. Okay. I just wasn't sure if I had cut you off earlier. So I'm going to tie this discussion with a question from Sizzigie and local. He asks, how likely is the scenario of designer babies or babies modified for athletic powers or mental abilities? And I want to tie this kind of the idea of gene drives. If gene drives for beautiful children, intelligent children, you know, longevity, long life spans, you know, who knows, maybe you don't get cancer, who knows? But those could be permanent. In other words, you wouldn't just design your child to be that way. But in fact, with a gene drive, you could, in fact, completely redesign the species to have those characteristics. And frankly, it just seems like who knows, I don't know, maybe not 100 years, but maybe in 500 years, you know, I just see, I just think that it's inevitable. I just think that is going to happen. I don't know how we can not use this technology to make that happen, frankly. But what do you guys think? Well, I have two thoughts, three thoughts maybe about it. One of them is that in general, I think it's not a good idea to mess with genetic diversity. But the second thing is it's probably not going to happen very often, even if it does happen. I mean, even if people who could afford to have their babies designed decided they wanted to look like X, Y, or Z. And the third thing is kind of strange. And that is that there's something in, there's something called the rare male phenomenon. It's about mate choice. And very frequently fruit flies will be attracted to the male that looks different from the other males. So if everybody looks alike, then whoever doesn't is going to be an advantage. I love this idea, frankly. But yeah, I know exactly what you mean. And that's true that there are studies that show that that is a true phenomenon. And right, that is a problem that if everyone looks like Second Life Avatar is basically kind of explains why the British Act can't think of his name. I do not think of as conventionally handsome, but apparently he's quite a heartthrob. Who am I thinking of? I don't know. No, not Hugh Grant. You know, he's docked. Oh, dang it. Anyway. Oh, I know what you mean. House. Yeah. No, not house. No, no, no. Who's the Avenger who has the mystical powers with the Come on, somebody here's got to know. No, no, no. No, he's one of the Doctor Strange. Thank you. Who's the actor? Who's the actor who plays Doctor Strange, the British actor? He's also Sherlock Holmes. Oh, is that Benedict Cumberbatch? Benedict Cumberbatch. Thank you. We finally got there. I don't think he's conventionally handsome, but he's a heartthrob. No. I think because of this principle that he's kind of unusual. But anyway, that was a long digression to get. Yeah, I'll make a general comment. And I think this is where we need to have a lot of good discussion of rational people that, you know, when you have some sort of new technology and you have new ways of doing things, we have this balance of what are we capable of doing and how useful is it versus what's good in society. And there's, of course, this balance of forces of people who have enough individuals and enough power and money and influence to alter what they want for themselves the way society handles things. And so, I don't know, I think, you know, the thing that's true about designer babies is that people would see those as valuable. People would be willing to pay money to have their offspring have better survival chances. That's a part of biology, a part of evolution. That's the desire. And yet there should be some role for government to rationally say what are the right things we do or don't want to do, especially in the face of a very good, you know, market thing that can occur. And it can be very profitable for some people. So, you know, I don't know, I think, I think in general, our moral bias right now worldwide is to say that we don't find those things to be good. But how much the small number of influential people might play in certain countries that don't necessarily have the same type of ethical structure around, you know, maybe rich, poor gaps are going to say, yeah, this is going to happen. So if it happens in some countries that's unregulated, then what are the countries that want to regulate it do? I don't know, but I think that's kind of where we are in terms of the discussion. I do think some of this is going to be inevitable. Okay, this is fantastic, but I'm afraid we're kind of running out of time. I'm going to add a little bit of moderator status to those with one final point. Because CB, so that with this kind of gene tampering that they're often trade-offs, you know, that we in mind, or we bred tomatoes tough or machine-picking and shipping and driving in grocery stores. Yeah, tomatoes are not what they used to be. That's right. And I think that touches on, I think, an important point with gene editing, especially for desirable traits is that we may encounter sort of linkages between traits. You know, for example, like those Russian studies that tried to domesticate wolves, and what they found was that as you selected for friendly wolves, what you got were curly tails and floppy ears, and that those physical traits were inextricably linked with the behavior changes. That's right. And so we may encounter very similar situations as we try to edit humanity, where certain traits are just inextricably linked with others. It could make the whole situation extreme. And on that note, I'm going to exercise my authority to bring this to a close. I want to thank both of our speakers for a fantastic presentation and discussion. All of our students for their really good insights. Yeah, very interesting questions. I agree. And I hope everyone will continue to attend these science circle panel discussions. I think it's been really fun lately. That I'm going to wrap my gavel. Of course, feel free to stay here and chat in local chat, but our speakers go so they can turn their phones off. But everyone is free to hang out and talk to them. Thank you very much. Wrap, wrap. I bring this session to a close. Good night, everyone. Yeah, so I'll say I did budget because the way my talks have gone in the past is, you know, 135. If anybody has, just want to stick around for a little more conversation. Max, can you stick around for a little bit more? Yes, I can. Let me scroll back. There are a couple chat questions there. And I'm just going to go back and reverse order. A good idea. We might have missed something interesting. Yeah, I think, okay, so Laura has a question about worrying about designer babies. What is becoming technology is the ability to analyze the genome and basically select from within a pool of embryos what you do or don't want rather than editing. And that's being done now. Yeah, that's being done now. I guess the question is whether, you know, which might have a bigger impact. You know, I think IVF, you know, ultimately it'll be a combination probably of the two, you know, but that I think this ability of analyzing genomes of multiple embryos, one thing we keep in mind is that full sequencing of whole genomes is still very expensive and still takes time. And so, and then the more embryos you want to do this analysis with, depending on how much of the genome you're trying to analyze is really important. Now, if you have couples that say both have sickle cell trait and then they're worried about trying to not transmit it and have a, you know, a homozygous normal child, this isn't being done already. And so I think in terms of, like, if you take single gene by gene bases that in terms of the alteration of the future human gene pool, IVF screening without any sort of editing going on in the first place will be the wider impact of the gene pool depending on how much you really want to look at. If you want to look at a whole genome sequence scanning that might not happen, but I do think that, you know, this is more likely, especially worse so that it's designer babies. Designer babies, you know, I think that's really probably a relatively rare phenomenon that requires a combination of being the arrogance of saying exactly that we do want this and that that's what's going to be better for our child and have enough money. They also don't understand those kinds of genes as nearly as well as we do some of the disease-causing. I mean, like intelligence. I mean, intelligence has a genetics basis, but it's not a gene. Correct. The basic fibrosis is A gene. And beauty, you know, these are Yeah, beauty, talent, you know, those kinds of things are not that easily analyzed, I don't think. I hope to answer your question, Aurora. And there was one more of that. I think Teal had it was really interesting. Oh, okay, so... Yeah, if you had a question you didn't get an answer, just go ahead and repeat it. It'll be easier to find. Yeah, so Jessica Fox actually asked a kind of a ponders a question a real life society of all females as a result of the SL genre of genital charades. Yeah, there's actually other technology that allows basically females to reproduce and not males. And they would only be reproducing more females. This is the trying to get the Greek term for it. But, yeah, there's certain ways there are ways that males are becoming dispensable because they can actually alter female somatic tissue to become sperm that you can then do vitro fertilization. So I do fear for males in society. Oh, I know there was a question that came up. So there was a question in the chat that in terms of think about diabetes as a genetic disease that, you know, insulin is a treatment option and that the technology behind providing insulin at cheaper and cheaper costs and with easier and easier injections has led to a point where you might say, oh, diabetes is a disease where we have tears or symptomatic leavements. We don't need to do gene editing. But I would counter that by saying there are still lots of individuals who miss an insulin injection have other health consequences of not having an insulin diagnosis. There are lots of things that stem from diabetes, so why do we be worth fixing? Definitely. Yeah. If you can find those single gene alterations then germline editing makes a lot of sense. Or even somatic editing. I'm really against the majority of somatic editing because it's just something where people keep going back for treatment and there are going to be gatekeepers to that for every single person who needs it. Whereas if you do the germline editing and again on the assumption that future fertilization and editing is as safe and cheap as I think it probably will be in 5 or 10 years that that's a solution to problem not something has to keep being redone. Well, as somebody pointed out a little earlier in the chat insulin is cheaper than it used to be but I wouldn't call it cheap. Does that look like aspirin? Yeah, let me also point out when it comes to diabetes as an example disease I think like 85% of diabetes that's genetic type 1 diabetes has to do with autoimmune diseases that are very complicated and hard to understand. But there are some examples of monogenic diabetes that are single gene causes. So let me say that I'm talking about those types of monogenic fixes. Yes. Yeah, I think that things that have clearly defined gene as primary focus I think those are I think those might be good targets for this kind of And I will say, let me just also reiterate for the audience that one reason why agriculture and I think not only do I work for an agricultural company but a field of genome editing is excited about the applications in agriculture is because you can make your altered plant and if there's any sort of off-target effects or insertion of material you don't want to your final product those can all be bred out. You can look for those and make sure you're not accidentally putting stuff through more than what you wanted to change. And so that's why plants in terms of genome editing is this really exciting area because you don't have to worry about, oh, we messed up or we altered things in the wrong way. It's one of these things where you are altering but then releasing otherwise clean things into commercial space. Some of those modified plant genes though do tend to get out. Again, we have the technology to re-sequence entire genomes relatively cheaply and say to the base pair this is what we know is like basically the foundation plants for, you know, some sort of product. So I think we have the compensatory technologies that we use to make sure something doesn't go bad keeps getting better as well. So are there any more serious questions from the audience? Any follow-up? Well, this has been fun. Yeah, good timely topic. Okay, well, there's some intellectual property. I think patenting biology is always kind of tricky. Yeah. I get it with CRISPR because that required a lot of actually modification of the normal process to, you know, to make it work as efficiently as it does. In terms of what Bergon was talking about, it may not have been particularly obvious. I guess there's one last question, and then I think we'll let it go. And it's not necessarily my serious question that Gadica was why you went to this field right, and I will say that I decided on this field. I'm old enough that I decided on the field before Gadica could influence anything when it came out. All right, well, with that, I think yeah, it was great. Okay, well, thanks for inviting me. I enjoyed it. Yeah, time-traveling biologist, absolutely. Quiet! Don't hit it. Yes, I'm really a transgender Mendel. Yeah, well, it opens him you are, right? That's true. I do have a Mendel avatar. All right, so, I'm gonna pop out of here, Chantal. I'll take back my slide presenter. Okay, well, Merry Christmas, or whatever it is you celebrate, and it's gonna be 2019 before we know it.