The Future of the Genomic Editing Revolution - Prof. George Church - CRISPR

so thank you for thank you for coming I think it's we're gonna have a very interesting evening tonight and I wanted to start off by telling you a little bit about the origin of this the idea for this this talk and what its what its leading to over the next couple of days so we had very wonderful interaction myself jennifer doudna with bill Hurlbut who's sitting over there from Stanford and realized that there was an opportunity to get together and discuss the ethics of of genome editing in a particular human genome editing and we wanted to do this in a puttin it with starting off with a very public lecture with someone who really represents a real leader and and thinker in the field and so we were fortunate to have sponsorship for this by the Templeton Foundation so we're very grateful to them and it's a really a great pleasure and honor tonight to be able to introduce our keynote speaker for this evening George Church so let me tell you a little bit about George he's almost someone that doesn't need introduction but but there's there's really a lot to say about him so he received his PhD from at Harvard in 1984 and then he joined the Harvard faculty in 1986 and I think I would I met him around that time when I was a I was a graduate student in a very different field but you couldn't help but be aware of George and his big thinking and his incredible ideas and in those days you know he is he was starting to do work on DNA sequencing and thinking about how you analyze organisms from the level of the whole genome he his name has really become synonymous I would say with the field of genomics he pioneered the first direct genome sequencing method and over the years he's made many incredible developments and discoveries that have led to technologies for genome engineering and genome manipulation he's the currently he's the director of a center of excellence for genome science he's advised the government on policies and biosafety personal genetics and precision medicine and he's authored over 400 papers and also a book Regenesis which a number of us we're fortunate to have him sign for us tonight and you know I think that he's someone who's just been a real inspiration over the last couple of decades in the field someone who's always sort of thinking beyond what what the rest of us are doing and thinking about in science and really trying to ask where are we going in the future and that's really the topic I think for tonight and I'm very excited about his title which is future human nature reading writing revolution I don't know quite what he's going to tell us but I know it's going to be very interesting George were delighted to have you here [Applause] well it's great to be here and this is my conflict of interest slide and if it's been up there for a while but if it goes by too fast it's on my website as well and transparency I think is an important part of we technologists who take an interest in the ethics side of these things and and I just want to make a special thank you here to this organization personal genetics education which was initiated by ting woo a close colleague and and my wife in 2005 and this this is where we were at the off Science Technology Policy OSTP and also Congress that same day giving what briefing not not lobbying but briefing on genetics which we do on a regular basis PG had also teaches high school teachers and they visit especially underrepresented and underserved communities are throughout the United States mainly and then they work with screenwriters for TV and films to try to get the facts straight so and this is a picture of map IDI where people will mark that they have some that they will take a test and show they have some knowledge of genetics this kind of fun anyway Jennifer was at the congressional briefing and and we were had the pleasure of being with her when she got her Gairdner award in Canada at which she then donated the PGA Tour's so we were extremely grateful to her generosity so I'm gonna start with a little audience participation we'll have some at the end as well so I think most of you I'm just I'm gonna take a guess that most of you feel that human genomics is not useful and and I will ask you to raise your hand if you have your human genome sequenced you don't have to have it with you but if you have it ever had a sequence raise your hand well that's actually quite a lot I expect from well-educated that I want the whole sequence sorry I mean 95 I'll settle for 95% since a dirty little secret is that no one has ever had their genome completely sequenced anyway so if I'm gonna make the argument that it might be useful and if what's it useful for I will make the argument later on that you need to know your genome in order to edit it that may sound shocking to some of you but these are some of the things that you can do it with preconception genetic counseling this is something that does not necessarily put embryos at risk you can diagnose very serious diseases like Tay Sachs there there are hundreds of them now that are highly predictive and actionable in the sense that you can marry somebody based on that information and that may seem a little unromantic to some of you so there are in vitro fertilization diagnostics and prenatal non-evasive testing and so on there's newborn testing which many people are shocked to find out that that their children and sometimes they Moute were genetically analyzed for up to forty diseases that are actionable usually something like dietary then there's what's called a genetic Odyssey where you have a child that has a developmental delay or some other problem and you go from physician to physician trying to figure out what went wrong and very often the parents blame themselves and just finding out what went wrong at a finding an explanation in the DNA is a great relief that to the parents even if there is not a cure it just means that they didn't cause it and so anyway the light.the the list goes on and I think there's an opportunity here for preventive medicine very often we're talking about the cure for this the cure for that what about prevention i'ma just give two quick examples this is a and you'll notice that I actually show pictures of patients and I name their names and you'll think oh how can he be talking to us about ethics if honest you know first slide he's showing John Lauer men as a patient and the answer is that we run a study where we have where we make sure that they are educated and that they know what they're getting into and part of that is if you participate in medical research the chances are that your data will become public and you will be reified in fact it's quite likely that all of your medical records whether you're in a medical study or not is already in somebody else's hands that you don't want because your medical records are worth 20 times on the black market your credit card and if you're interested I can explain why so that's and so in this case John Lauer Minh was a an author for Bloomberg and he discovered in the process of our study Personal Genome Project that he had a jak2 mutation which was not germline it was it was in his blood and he and it turned out that that he had reported that he was healthy but we we found out in his medical records that he had scotoma as in his eye and leg pain and so he takes aspirin for life not too bad here's another one who you know really did a wonderful thing for the world in being quite open about her brca1 genetics and then get and then talking about the decision about whether to trust the genetics and take a preventive met medicine here a lot of people people think that Angelina Jolie had a lump or some positive result in an x-ray this was entirely based on her genome she was said to have 87 percent chance of breast cancer before she had surgery and less than 5 percent afterwards so that's an example of preventive medicine not reactive now this is a list that you'll see a few times during this talk this is my personal ethics list these are dated in the order they appeared in my life they're not necessarily significant or logical in any other way and they represent ethical things that I've dealt with and I teach a course in ethics by the way it's a required course at Harvard Medical School for the graduate students and we talk about some of these things and we're not going to cover all of them today but the first one I want to talk about is and it's been throughout my career not just in 1974 is this idea that when a scientist stands up here like this and talks to you and talks with animation let's say about a subject that means that he is or she is advocating that subject that he is an enthusiast if I were to talk to you about let's say Neanderthals then you will all conclude that I and have an active program to clone the inner Thals this is not this case or should not be the case I should be able to discuss topics that I think from my point of view are alarming or in it possibly in a negative sense and I can say it with animation without being an advocate or an enthusiast and so to that end I have been co-author on about 30 some articles on the topic of ethical legal social issues something that you know I've been very privileged to have very close relationship with Jean tain once off for example who is embedded in our laboratory as a as a bioethicists and very often I we seek solutions to the polarization that often happens in these where you were looking for a win-win where a week and you'll see some examples on this list as we go through them for example that you know for the the win/win for embryonic stem cell pluripotent stem cells was induced pluripotent stem cells that was something that satisfied most of the argumentative groups on both sides not every men so let's talk about equality as something that often comes up at the end of a long meeting like this one that you know Saturday afternoon it would come up well what about equality you know all this great technology and I'd like to have this conversation start with some of the tough problems and so let's start with that and of course I'm going to give you quirky view on it and take it for what it's it's worth so what is the revolution we're talking about here I said reading writing and revolution you could say that's that's my version of math but what is the revolution here and of course we all know the revolution is crisper but I think that the I'm not going the normal acronym none of those letters have anything to do with the CRISPR technology that we're all enamored of that we're all fixated on it is so I'm gonna give you a couple other words that I think better capture what the revolution is and it has many components not just nucleases but the idea of comprehensive data biology or molecular biology used to be the anecdote the single gene single base pair but now comprehensive data is a possibility and often insisted upon we have recombination not just cutting but actually cutting and splicing and making exactly what you want we have informatics we have sequencing we have pluripotency that is to say that you can go from a single engineered DNA molecule into a whole organism plant or animal and we have reduced costs greatly reduced cost and that's what the next two slides are about and for you know for some of these things I know this broad audience some of these pictures will be you know inappropriate they will be complex just think of them as eye candy so I'm not even gonna explain what I just showed you but I like the Novus seek instrument that's the newest of the next generation sequencing it's named after my granddaughter whose name is Nova anyway it's just kidding uh-uh so her name really is Nova but anyway we have this exponentially shrinking cost of reading interpreting your genome so after I had been obsessed with this for a long time in fact I've been obsessed AddThis ever since I left sung-ho Kim's lab we who's here in the audience when I was 20-something years old and and shortly thereafter we could eat before that we couldn't even conceive of the project but when we finally started delivering it looked like there's going to be three billion dollars which is about the price of this tallest building in the world and then it's come down ever since then it's been about a thousand dollars for over a year and in a certain sense it will be free soon in the same sense that that phone has on it free software you Google Maps they used to charge for it and now it's free and then in fact many people in our project get their genome for free and so this is the these are the curve sees the more technical versions occurs and the the thing that is most striking so this is factors of 10 on the y-axis and it goes back to almost the time that I started doing molecular biology and it was on a Moore's Law curve which is a very very steep exponential for computers it's going very fast but then it got even faster in 2003-2004 so what was going on there we'll talk about that in the next slide but this is for both reading and writing DNA I would say that reading has improved over 3 million fold during that period of time most of it in the last few years and writing a short oolagah nucleotide says improve by a billion fold over that same period of time a billion fold and I think that both of them could improve by another maybe a thousand to a million full don't ask me when and don't don't remind me when it doesn't happen but anyway that's my guess and so far we've all heard under predicted it rather than over predicted it so what happened in 2003 2004 I will give a totally self-centered and self-serving synopsis which is that these two papers we figured out how to miniature multiplex and self-assembly could be used and biology should be really good at this sort of thing at it is atomically precise which is about as miniature as you can get and it does a lot of self-assembly and so we just captured that along with the revolution and bio and story and Nana in electronics industry of micro fabrication in both reading and writing and the the key thing here to take away is you can't do one without the other our our most popular sequencing method is literally called sequencing by synthesis and every time we do synthesis or editing of a genome the clued end people do know the genome that they're editing you shouldn't be editing a genome that you don't know although you can you know just like you can you can text and drive a car at the same time there is just as I've been impertinent enough to redefine CRISPR um be blasphemous enough to say that there is more than one editor in the world there are nine editors and what editor these molecular machines do is they scan your genome which for humans this is six billion base pairs looking for one place that they like 120 mer 20 ACS use in 200 a row that the Machine likes and making either a cut or a combination at that point how does do that how does it fine one and six billion well the scan head the the place where the action occurs where that recognition of 20 base pairs out of six billion occurs is either DNA in red or RNA in in blue which which is what CRISPR uses cast nine or therefore these protein to do the recognition so you have watson-crick base pairs that are doing the scanning for DNA and RNA and proteins have more complicated but at this point very well understood rules and you can you can dial them up and so that brings us to the next question why is everyone so infatuated with CRISPR why do we think that that's an improvement at all and a lot of people say well it's really easy to use in its low cost and that's the reason that we have exalted it to you know such a high status I will say that maybe and but maybe not because about about the time that we were starting to publish this deluge of crispr papers were the first few had come out this lovely paper came out from Junsu Kim's lab where they with with very little fanfare assembled an alternative CRISPR nucleus that so-called Talon accounted for all of the almost all of the protein coding genes so eight over 18,000 of them so it can't have been that hard to do even though it wasn't CRISPR I realized this an anecdote but there you have it that though that was the previous technology and its day in the Sun only lasted about a year Christopher has lasted now of Wapping you know four years in the Sun and it may it may it may last forever maybe the only edit that maybe the last editing method but this is kind of the history of I could write the same thing for genome sequencing a very complicated plot of a whole series of replacements but in this case it starts with homologous recombination in mammalian cells with Oliver Smithies and Mario Kikuchi and some people ask who's gonna get the Nobel Prize for genome editing and I say it's already been given to Mario capetti and Oliver Smithies I mean that's very silly of me to say so but but the point is there is some progress but there's also a lot of you know this is again factors of 10 on all axis there's a lot of constancy in the editing efficiency but I think it's the editing efficiency that is what we are celebrating here with CRISPR now to be a party-pooper again there is if you're going to be applying CRISPR especially in a clinical setting where you're where you're treating a large number of human cells simultaneously if any one of them gets an off target event and most drugs have some kind of off target event if you have an off target with CRISPR and it lands in the middle of a tumor suppressor gene then you you run a risk of getting a tumor and so you want the off target at least in two men suppressor exons to be as low as possible and I'm not going to go through them but there's a whole bunch of and this list doesn't isn't even give it justice there's a large number of articles that have improved the specificity of CRISPR at least in certain stat contexts and one of them even claims from my group that that you can detect single nucleotide polymorphisms this jargon snip you can detect you can make a CRISPR that will up recognize one be off by one nucleotide and you can do this fairly routinely it was published in you know the first ranked Journal called by archive and I urge you all to publish there and you'll see a few other references like that but even as we bring down the the off target as low as possible the more changes you make the more risks you have and they multiply out so even if you have an error rate of 10 to the minus 4 let's say which is among the lowest that you can get one error in in 10,000 cells if you're treating a billion cells that's a lot of errors so that's off target errors what about on target errors these believe it oh these are ethical issues ok these are these are safety and efficacy issues most mostly safety but on target CRISPR is not ideal in fact none of the nucleases that have been used the mega nucleus is the talons the zinc finger nucleases because they create a race situation a race between repairing the double strand break and hoping for the best where the cell makes a mess which I call genome vandalism rather than editing and what you want very often which is to make a precise change again there are solutions to that and my two favorite are from bacteriophages in fact CRISPR itself is something that involves bacteriophages most of the gifts that on slides I've been mentioning most of the technologies were not invented in the sense that we thought about the atoms from first principles and said oh I'm going to design a CRISPR they are gifts they're gifts from the microbial world and these two are as well and these work the specificity that we want on target by not making a double strand break but by only bringing to you only once they bring together the donor DNA that you want to change and their target then they do their job and these are not these are not as applicable as CRISPR you don't go out and sell your CRISPR stock they they are very specialized in fact one of them the beta recombinase here only works in e.coli k12 you might say that's coincidental coli k12 is my favorite organism I'm sure you're all saying that right now even the non scientists you love e.coli k12 but anyway but we've done arguably one of the world's largest and most radical genome engineering and I think in this case radical is a good thing maybe where we changed a four million base pairs in ohm to eliminate one codon so all of the organisms in the world synthetic and natural use 64 triplet codons so ACGT to the third power except for this one this one only uses 63 and we expected it to be useful for four things and in fact it is useful for four things it uses non-standard amino acids very efficiently meaning we're not no longer restricted with the amino acids of nature we can put in completely chemically synthesize amino acids it is genetically and metabolically isolated so here's an ethics lesson and I hope you haven't missed this is biocontainment if we're going to release something into the wild we want or if we're gonna make any new technology we want to have a reverse button we want to have we want to have some ability to to go back and so that's what this is and we've proven it and then finally multi viruses and I think that's very profound sort of philosophically as well as practically which is the idea that you could make an organism resistant to all viruses including viruses you've never studied and that you don't understand as long as they follow one rule which is that they use the genetic code of the host if you change the genetic code of the host they say hey what's up it's not working you know and they can't and it's radical enough that they can't even evolve around it that's a theoretical prediction and it looks like it's turning out to be very very likely we were surprised how multi virus resistant these organisms were with just one code on change and now we're doing seven we're gonna jump now from that radical recoding that radical application of something that goes beyond editing we call it genome project right variation on genome project read which is the first genome project this is writing the whole genome beyond editing we're gonna change to engineering humans we're going to go into this gently we're gonna start with something that that's probably not controversial which is engineering human cells for Diagnostics for their testing therapeutics for testing hypotheses when you we started this as exercise as to why you would want your human genome if you get your human genome some things are highly predictable but other things are variants of unknown significance if you get one of those variants of unknown significance you'd like to know how do i how do I make it determine whether it's harmful or not and this is a new way of doing it which is you can alter a genome by one base pair or however many of the difference is let's say well as little as one and then you can make it into a complex tissue or organ or you you know modestly called organoids and then test whether that the function has changed you can test cause and effect you no longer need a cohort of 10,000 to prove you know you don't need to collect gigantic human population samples to get a correlation which is not convincing you can do cause and effect as long as you have convinced yourself that you're changing one base pair and that you've got a good organ model doesn't involve animals it involves actual human organs and so in this case we start at the very top there with PGP one which is code name for personal genome project which I've already introduced number one full disclosure per the the individual number one and I think I said I can release the names of the participants is not John Lauer Minh but it's me and so we sometimes call me guinea pig number one GP one and you take those cells switch your fibroblasts turn them into stem cells turn the stem cells put in the crisper cast nine and then use that with a repair all ago to to take out that one G if you leave out the repair all ago it makes a mess that vandalizes the DNA but if you put that in you can change just that one G and and you say well I've been talking to you about off target on target messes you expect me to believe you just change one G in the genome out of six billion and the answer is we sequence the genome this is a clonal cell line so this is embryonic stem cells here's an important lesson is that even if you have off target if you have a clonal where you've grown it up from a single cell you can sample that clone and show that it is extremely unlikely have anything off target in this case this baby was hypothesized I mean in some cases you might have hundreds of hypothetical changes in this case the most likely one was missing one G on a sex chromosome boys have only one X chromosome in every cell every nucleated cell and in this case we wanted to test that so we made two cell lines from PGP one that went differed by that one base fare there's a clean experiment if we just compare yourselves to mine or this baby's to mine there'd be 3 million differences that's not a clean experiment but changing one base pair is and so Lu Han yang who did this work thought nothing didn't even bother to consult me as to whether she should sequence the genome and that's how we route what we routinely do we change one base pair sequence all six billion okay and so then this is AA just example of these cardiac like tissues with the beautiful saw repeating sarcomere organization when you have a normal stem cell again from PDP one on the far left here and then change one base pair and it's abnormal physiology biochemistry and morphology so you can determine cause in effect and if you run this in Reverse you have gene therapy so now we get to more serious that was genetically modified human cells and human organ nodes this is you know actually genetically modified humans and some people are surprised to hear that there are genetically modified humans running around these are there are 2,300 clinical trials research trials on gene therapy there is only one approved ironically in Europe where they're not keen on eating genetically modified foods but they're okay who is uniquely modified humans and this is the most expensive drug in history it is a million dollars a dose now to somebody like me who likes bringing down the cost of things by a million to a billion fold this is unacceptable and we have various strategies that you can we can talk about how to bring that down the main thing is increasing the market size but the problem is a lot of these are orphan drugs they are very very tiny market size but there are some like infectious diseases that will have gigantic markets and that will bring the cause so now we get even edgier so he went from human cell engineering to organoids to human gene therapy which typically is done in adults or in some cases children you have to do some cases the earlier the better because for example if you cure blindness which is a real thing it happens in gene therapy you have to do it at a certain age otherwise they'll see photons but they won't be able to see faces and processed faces and you can imagine taking this further and further back you would want to engineer in utero or earlier so what is the status of human germline therapy some people take it for granted that this will never happen it doesn't happen anywhere and it is not allowed anywhere that is very far from true it is permissible in many countries including the United States human cloning is actually permissible in the United States but the question is not just what is permissible whether it's acceptable and it is already done so again here's showing a real real patient this girl in the middle had mitochondrial germline now some people say oh that's not really germline well but it is it's passed on to her children most of these therapies are to reverse a problem and I think you can either think of it as a slippery slope if you think in negative terms or as the way of carefully testing without putting too many people or embryos at risk is it will start with it will it will move on from mitochondrial therapy to sperm infertility or you'll be reversing you'll be changing men with certain kinds of infertility genetic traits to fertile and if you could do that by just engineering the soma the somatic cells around the sperm then that would be considered ordinary somatic gene therapy meaning not affecting norm line but since it is a sperm itself this effect in these then you have to at least reverse it now you're just changing it to what everybody else has but that will probably happen and if it happens in clones like the clone that I showed you your cardiac muscle you could in principle get something that's close to 100% correct Nords it has all the low error rate of crisper plus all the low error rate of checking the clone by sequencing so this could possibly make it in the fertility clinics and then if that works well no embryos have really been put at risk there then it would be but any more so than any other medical procedure and then then you could move on might the world might move on with due consideration Duke conversations like the one we're having here two serious diseases like Tay Sachs which I said right now is handled with either abortion or in vitro fertilization where abortion you might sacrifice 25% of the children that are that are born with the recessive disease like Tay Sachs and with IVF you might sacrifice more like 80% of the embryos so some of them will be normal but they won't be implanted and for a large fraction of the United States and the world that loss of embryos is unacceptable so this might be so usually the way this conversation is phrased almost every conversations I've been in is we are putting embryos at risk in this scenario where actually could be saving embryos putting fewer embryos at risk than current medical practice so I just put that out there and then finally also note that now there is great progress in making eggs and sperm entirely outside of the body mostly done in rodents so far so let's okay so we've gone from human cells human organs adult humans therapy germline therapy which is past tense now let's talk about enhancement I'm not doing this just to be provocative I think it's important to visualize things in advance even if they never arrive we need to talk about it and I'm gonna make the argument so here are some of the traits that you might want to enhance in human beings or not theirs it might be the sort of thing you want to prevent the world from enhance we right now can only see a very tiny sliver of a visible light with our eyes we can only hear a limited range we can only sense certain chemicals and the list goes on touch each sensing in our memory is very limited we locomote very slowly and so forth but we are already augmented we are almost unrecognizable to our ancient ancestors our ancestral limits have been blown away we can essentially see the entire electromagnetic spectrum from gamma rays to radio and the list goes on I think you get the point we can go so fast that we can escape we can reach escape velocity from the earth and then it survive in the vacuum of space and the extremely cold 15 degrees Kelvin out there we have greatly augmented and in fact our goals like being able to go into space and survive and the vacuum and cold would be incomprehensible to our ancestors so I will argue that most of the augmentation that already that will exist and already exists is physics and chemistry it is not genetics if we are to talk about genetics it probably will be intelligence immunity and longevity or aging reversal and much of the ethics that we will talk about will we may talk about other things but if we get to something that's safe and effective there's a tendency to drop the conversation for example in vitro fertilization was considered very negatively it was described negatively with terms like test-tube baby Justin baby doesn't sound as negative today as it did back in the in the 60s but it is but when Louise Brown was born and she was healthy and beautifully healthy suddenly the FA turned around almost 180 degrees where it went from we should never create the monsters of test-tube babies which could be teratogenic like too we should not deprive parents of the of their right to have babies of their own that is the product of the union of two people that love each other but there are things beyond safety and efficacy we might drop them if safety and efficacy is proven but we need to be very cautious and these include the possibility that parents might treat their kids like commodities they might say which in the services they already do they get them the best education and they expect them to perform there are parents that will be making choices where they choose to have children that have hearing or do not have hearing that have particular genders in fact this is already if you look at in vitro fertilization in the United States a very common practice is to choose a gender and guess what gender that is 80% of the time female and I don't know what your expectations were but I was a little you know it's it's it's there and I've seen editorials who give reasons we can talk about that in the discussion and the other thing that will happen that we need to be very cautious about which is not safety and efficacy is this loss of neuro diversity if you look in a classroom there is a great desire to turn the classroom into an exercise like Henry Ford's of mass production where all the children sit in neat rows dress the same way they showed diversity in their skin color but no diversity in their behavior and appearance and if if somebody fidgets too much or falls asleep this is a bad thing you should medicate them and believe me if you could genetically medicate them that would be even that would be that would be something we need to be very cautious about because some of our most amazing citizens are those who are on the edge of some spectrum or other I think we don't want to lose them we might want to allow them to not be in pain some of the time maybe to turn it on and off but not lose them and it's very hard to - we talk about diversity but do we really mean it okay GMOs how many people here shop at a market that has the no GMO rule I do geez this is you're not a radical Berkeley crowd here okay or qb3 whatever I will argue that there are some GMOs that nearly everyone likes even the people that are anti GM foods and here they are it's a longer list in this but I picked a few here that that are recombinant proteins that treat some some of them fairly common diseases others are orphan diseases so there is a difference of opinion when you get the things that are extreme health interest and sometimes extreme economic interests for example Hawaii they banned GMOs from the entire set of islands except for papayas because that was they the papayas would have gone extinct so there was some room for negotiation on that subject I would argue the future of GMOs and this is actually a report from one of the non-gmo project groups is that one of the main problems with genetic engineering is that that you insert genes randomly and that could create toxins and allergens well actually random mutations are definitely random and we're getting better and better at engineering that's not random so I would argue that if you want to open a car door you could shoot it with a random shotgun fire or you could engineer a handle and use it so so we get two very interesting definitional issues here this is not technology it's possibly ethics which is sis genex versus transgenics now transgenics is a pretty commonly used word and then almost is the definition that people use for defining GMO GMO plants and natural and organic is that you have moved the gene between species over a great distance and the distance is actually defined in regulations how how far apart the plants have to be let's say sis Jenica is a much less frequently used word so far but I think you're going to see it more and more because it is a regulatory if you think of it generously is a regulatory opportunity or a loophole if you think less generously but there are 30 GMOs already in the last five years alone that that get through the USDA and to some extent international regulations because they're essentially they're changing like one base pair you change one piece here that could happen in nature and it's very easy to tech the transgenic because you got this whole big chunk of DNA that came in from a bacterium or some other plant but if you change one base pair that could have happened by the shotgun approach and you just cleaned it up by conventional genetics but and one of the famous recent examples is the white button mushroom where they knocked out a particular gene in order to prevent browning but an example of the transgenic which surprisingly to me should have passed should have gotten a pass from the critics because it is a matter of life and death it is not a typical that the reason that the anti GMO forces are justified for foods is it really isn't a matter of life it it's not even a matter of taste in most cases there's some subtle thing off off campus having to do with farmers but Golden Rice actually affects millions of people I mean our sorry the thing that's addressing which is vitamin A deficiency millions of people go blind and I they they often die within a year of going blind it is a cause-and-effect relationship and Golden Rice however this was started back in if it was working by 2002 it started well before that the decision was made to make it transgenic it didn't have to be transient I think I can't prove this because beta carotene which they're making is made already in rice it's made in the wrong place in rice and so sis genic Li you could move it but instead they imported two genes from two other organisms one of them a bacterium or wynia so that's I don't know where that's going to go I don't know where Golden Rice but it's an example of transgenic that did not escape even though it's a major health threat now here's something that is you could say it's not beyond safety and efficacy it is part of safety but it's not what the FDA the USDA and the epa usually worry about they worry about sort of like the distant future which might be next quarter or maybe ten years from now but this is a hundred years and here's the juicy example and hopefully many of you know that in 1872 Yellowstone was established and the gray roof grey wolf was already in decline both in Yellowstone and elsewhere and it was completely gone by 1926 the Endangered Species Act was the second such act and it allowed reintroduction many years later in 1995 it began and the impact of the president of the reintroduction of wolves was dramatic they the Elks didn't like it they didn't get a vote but they started killing a couple dozen elk per year that result in the Willows coming back which resulted in the beavers coming back which resulted in otters minks wading birds water fowl fish so forth it was a really amazing impact on the environment so we made a mistake back in 1872 not established in Yellowstone but getting rid of the wolves what if we're doing similar things today but it's not sufficient to say oh well well let's not change anything because that can have negative effects as well so let's talk about some some quicker examples here here xenotransplantation so this is even worse than transgenics if you think transgenics are bad this is moving organs from humans into pigs and then pigs organs into humans and this wasn't something like this was in the news today and it was also in the news many times here's one beautiful article by Carl Zimmer on the topic we have some skin in the game here but it goes way back before I you was interested in it which is this humanization of pigs goes back at least two decades about 15 years ago there was a a billion dollar investment in this field and it died not just what there was actually a pretty good road map to changing multiple genes I think they thought it was one or two or three genes it's now probably 50 genes that need to be changed or more but they had a road map what freaked them out was that the pig organs were producing viruses that could infect the immune compromised recipient of those organs and and that would be a bad thing probably it would you know you don't want to have swine flu crores the equivalent evolving in your immune compromised patient so so when we got the awesome power of CRISPR lujan who is one of the co-inventors many co-inventors she decided to try she and her team decided to try this getting rid of all 62 and not just retro viral genes from the genome of the pig all at once with one CRISPR and it worked it was actually pretty easy we were surprised to how easy it was up to that point people were doing one or two maybe three genes at a time 62 just seemed completely out out of out to lunch but in 14 days sitting in the 37-degree incubator that's all it took and then a little bit of PCR to screen it and a thing that and and so now there are there are piglets that we have ultrasounds on these things where many many dozens of other genes have been changed as well and I look forward to seeing and hopefully holding these piglets when they are born but if they're not we will we will try again and what excites me about this is maybe a little more subtle than the idea of curing the transplantation problem which is very acute it's not just that you and I aren't compatible for exchanging organs it's that there just aren't enough of us to give organs but it's more than that which is that would we produce organs we are going to be highly motivated to do preventative medicine which is to make organs that are pathogen resistant cancer resistant and Aging resistant we are not quite so motivated to do that in humans directly it is it was very hard I think to get FDA approval to take a healthy human and try to make them resistant to pathogens cancer and aging that preventive medicine is very hard to do unless it just involves taking walks into eating broccoli not something as radical as gene therapy aging reversal is is a real thing and we will see it both for physical as well as cognitive disorders because we have an aging population and it will be extremely important it I would imagine that many of these things will be used off-label to reverse aging and people who don't have any disorders that are not disabled do not have cognitive disorders but here's an example there are two actually two classes of example of aging reversal has been shown in mice one of them involves hooking up young mice with old mice they're there blood systems I don't recommend you do this with your children but but this works and another one that's just recently been published is using the same reprogramming factors we use to establish pretty potent stem cells so you can do that on a whole organism basis and it reverses Aging gene drives gene drives was something that was hypothesized again about two decades ago actually was observed in the 70s by bernard dijon who was one of my mentors as a graduate student but then hypothesized that you could turn it into a technology by austin burt but it didn't go very far until the awesome power CRISPR came along and Kevin s felt and indeed spied ler here a graduate student in my lab and shared with familiar Uchiha published this paper where we didn't do the experiment we did we didn't do what they did with gain-of-function flu viruses which was to to not talk about it until it was ready to publish we talked about it before he did any experiments to see if there was a gotchu that people told me and to talk about ways to reverse it and it can do all these amazing things you know reverse invasive species and reverse herbicide and pesticide resistance and most importantly control vector borne disease such as malaria and Lyme disease nobody likes lyme disease or malaria and what it does is unlike normal inheritance where half of your offspring will get will inherit the trait here 100% of them inherit the traits so it spreads exponentially through the population and does this time does not permit a long description here but it does this by using CRISPR which is mechanistically we you know if scissors it's basically scissors yeah the crystal structure indicates that they're scissors and and we put in many different scissors up to 8 so far in the mosquito experiments we're doing to make sure that the the or the mosquito genome doesn't become does not become resistant the CRISPR gene Drive we're essentially creating what would normally be called selfish DNA but in this case it's being altruistic it's doing what we want which is to carrying the purple cargo there purple cargo is resistance to malaria we're not trying necessarily the wipe out the mosquitos though some people would like to do that but to make them resistant to malaria with a whole variety of known anti-malarial antibodies and in small peptides and those scissors are aimed at an essential gene a ribosomal protein gene for example so that if it repairs by genetic vandalism it dies but if it repairs by copying the gene drive making two copies it lives that's the base that's you can now tell anybody in the elevator that you understand gene drives I hope if not ask me but now here's the sophisticated stuff on the top is how fast it will spread this is mathematical modeling which has been confirmed by experiments in some animals sorry some organisms and then we have a new one which is again published in this amazing high-profile journal called bio archive where it's called a Jay Z Drive where we have three drives CBA and none of them are really gene drives in that they cut themselves they cut the the part of the genome they come from they cut the part of the genome that the next one comes from and this just trust me on this or read the paper the yellow decays because it is slightly deleterious and it has nothing driving it the orange lasts a little bit longer because as the yellow driving it and then the blue has yellow driving orange driving blue and it persists longer but this allows you to do geographically and temporally controlled drive so we have reversal and we have reverse and we have drives that are local okay so what if we cure aging and eliminate poverty and diseases of developing nation we're gonna have a problem which is overpopulation or at least some people say that and I stand and I don't think it's a great solution to say oh well then we're not going to cure diseases of poverty or of aging which is a disease of industrialized nation so one possibly is going to space and I don't mean this frivolously it is a possibility and it's a good idea from our species standpoint because we are at risk for super volcanoes and asteroids but out in space we have a new set of ethical problems which is space radiation even on Mars the gravity is very low and then there's space genetics issues so we have a consortium on this and here's some of the challenges we have this gravity Austria prosess there are all sorts of neurobehavioral issues the radiation issues microbiome issues I've made a list of kind of a quirky list here rather than rare disease causing genetic changes these are rare protective alleles and I won't go through the list but they're things that make your bones extra strong maybe that could could result in something that would help with osteoporosis in space or on earth and there's some that reduce sensitivity to pain which you might be able to turn on and off rather having you don't want to have it off all the time because you end up hurting yourself and not knowing it so little kids will like to on their tongue and things like that and then there are there ways that you can ABC 11 will give you low odor which is very important in closed space where you're all together and in fact is very common allele among Asian populations of the good version that is and the list goes on and down at the bottom you start getting into things that are so rare you don't see them they're actually synthetic and they've been tested in animals and they include things that have locate cancer and high ability at cognitive tests what about radiation resistant this seems really improbable but here's a case in the literature that where you can improve radiation resistance up to a hundred thousand fold with just four mutations and and there's a wide variety a ssin among naturally-occurring organism but this is a nice experiment because it's isogenic there's the only difference those four mutations and then I'm just going to end on this very speculative notion where the normal surgery ward we have a choice as to what we're going to take into space are we going to take the entire Noah's Ark including you know the Giants Hoyas in the and the whales and the malaria and smallpox and so forth or are we going to leave something behind most most notably germs and if we do then we don't need to wash our hands for surgery and if we can turn pain on and off we don't need anesthesia so I just want to thank a bunch of I'm thank the few along the way but there's these people that have helped in our center of genomic engineered organs and we have some brain initiative grants and the these are photos from the annual get conference for a Personal Genome Project where the people whose identity we should be keeping anonymous wear name tags thank you questions or discussion actually it looks like it's locked on maybe it's on this on great so George Wow that was wonderful um so what I think we're gonna do is we'd like to have a have a discussion and we'd like to invite questions from the audience and we have a couple of people that are running around with microphones so if you have a question if you could just put your hand up and we'll bring you a microphone and we'll open it up to the floor so I see I see one over here thank you so much so as a precision medicine continues to grow and we continue to use genome editing technologies such as CRISPR what sort of implications does this have on the cumulative effect of off-target mutations and say non-coding regulatory regions for example if an off target deleterious mutation occurs in the germline new pathology is that we haven't seen before and may arise and perpetuate throughout the population alright wait before you give up the microphone clarification are you talking about off target trying to fix something yeah if you're trying to fix something yeah and we don't know what sort of off target effects these genome fully these you know editing technologies have what are sort of the ethical implications of that yeah so this is ethics in the category of safety and efficacy safety mainly I mean the safest thing would be to insist that you have nothing off target at least with it detectable which means you get the the CRISPR as low as possible mainly due to computer analysis and then and then you sequence the clone now most gene therapies are not done on clones they're done you'll put you'll you'll treat a billion T cells for example ex vivo and put them back in the patient and so every one of those is an independent event everyone has a chance of being off target the most serious things are probably giving tumor suppressor genes and it probably gonna be in tumor suppressor exons but not all of them and I guess it'll be like every other drug is that will go through phase one two three and even four phase four is in the public and well when you start finding things that actually cause cancer then you will go back and fix it but it will be very hard - it will be very hard to prove it you certainly can't use animal models because they have a different backgrounds you know which is the whole point it's off target in a genome that is not a pig or a mile steno so it's probably gonna have to be tested in in humans but if you want to know what a particular base pair does in terms of cancer formation I would say organoids are getting better and better and then we should exploit those as much as possible but right now organ I mean if it's something that happens in 10 to the ninth cells organizers are pretty small they're they're limited to about a half a millimeter we're getting we're starting to get vascularized organoids and those will be whatever size you want so okay I see a hand there Thanks you mentioned in one slide that one way we could get to Janette germline modification in humans is through synthesizing new eggs or sperm from you know embryonic stem cells and the like but there are a lot of you know mutations that are occurring in these attempts in mice so far and I was wondering how you see this progressing how do you think it's doing and do you think there's a lot more work to be done before we see that working safely in humans right so the mutations you're probably referring to our patients occur just from growing cells so if you look at your body it's full of mutations almost no two of your cells have the same genome in fact a lot of your cells are unemployed they're missing or have an extra chromosome so it's a lot of changes in your body that doesn't mean that we should glibly start creating germ cells that have the same kind of me you have in your body in fact the standards should be much higher but it is true that a huge fraction of human births never make it to term or sorry human conceptions never make it determine probably a lot of that is that they have mutations that are deleterious to embryonic development so I you know I think it's a huge challenge I mean you hit the nail on the head it's not due to CRISPR it's due to just growing cells growing cells have a mutation rate of about one or few mutations per cell division and it's a little worse in Colt cell culture than it is in vivo but it's a problem both places so it's an open question but hey that's the fun of being a genetic engineer in this in 2017 so maybe I'll just interject here and ask you so I think you know during your talk you presented a number of different areas where genome editing and engineering is going to have a big impact in the future and and I wonder what you what do you think is the is the first you know area where we're gonna see this kind of impact is it going to be in transplantable organs is it going to be in a clinical benefit to patients or maybe something else well here's here's where I can cheat by saying that just focusing on what we already have so we already have genetic genome editing for ccr5 and treating patients with HIV infections that already have HIV in fact not preventive and that's done with zinc finger nucleases we also have treatment involving car t-cells or cancer therapies so it's it's a pretty safe prediction that those are at least going to make it through serious clinical trials well some of them are already in Phase two clinical trials I think applications to infectious diseases beyond HIV is going to be a major place where you can get larger populations so that's a fairly safe that as well but you know the the orphan drugs have been the northen drug Act has made it affordable two companies by getting reimbursable huge you know a regular orphan drug might be hundreds of thousands of dollars per year and if you can do that with one dose and that's that could conceivably save money so that I think that's where we're going yeah I see thanks so much for coming here George I was super happy to see animal rights on your on your list there but then I was a little confused when you brought up sort of growing organs in in pigs I was surprised not to see sort of a straight jump to like in vitro organ growing is there is there a reason for that it seems like some sort of conflict so yeah that's a very accurate well I mean observant people wonder how I can be a vegan and also allow one of my postdocs to go off and start a company on making transplantable organs you know a fairness we are trying very hard to get organs to develop in in the lab I showed an example of an organ I'd which is a cardiac muscle and I think we're one of the first labs that has vascularized organs growing in the lab and and we that allows you to pump blood for them and get larger and more realistic ones I'm just worried that if given a choice but if everybody stopped eating pigs tomorrow for all reasons that would or hurting them that would be great but this is a drop in the bucket compared to bacon I'm not wild about it you probably aren't either but it's a it's a temporary we don't know if we can deliver organs accurately entirely in the lab I think we can but until we're sure that millions of people are dying for lack of organs it's it's a very it's a example of a very tough ethical decision of whether a pig's life is anywhere close to to human life and and that pig would what could save ten human lives because you've got all the you know heart liver lungs kidneys and test and so on I am very I apologize in advance I see one in the hand in the back there doctors in illegitimate analogy to say that aging and cancer and some of these diseases are the Yellowstone wolf and humans are the Yellowstone elk and should there be any of the pathogens or something that should be not addressed prevented etc by genetic manipulation yeah that's a excellent question in fact there are experiments that so some people feel that that you need we've got to this sudden infatuation with the microbiome there's a lot of microbiome products I'm involved in five microbiome therapy companies but the fact is we don't need our microbiome for life there have been almost every animal ever experimental animal goats chickens and even humans have grown without any noticeable microbiome they nevertheless they're very helpful so you can you get certain less than lethal diseases that can be cured with careful carefully chosen species of microbes there and and for awhile people thought it was just good bacteria so-called good bacteria but there are actually now shown that you can do some of this also with viruses like norovirus so I think we have deep ignorance that hopefully will be fixed very soon as we move from merely observing the human microbiome to doing therapeutic experiments and experiments on germ-free mice but it's it's it's a great question and wealth well put in terms of the wolves maybe you could edit the human microbiome well you can you you can edit the human microphone but I think there's a assumption that that is pretty healthy out there you just have to get the right one it has been shown that you can confer obesity and diabetes be a microbiome to germ-free mice so you want to be very careful and even with naturally occurring microorganisms hi you discussed human enhancement through genetic modification and I was wondering if you could give us like some examples of what is in our reach right now and which kind of human modifications would be more challenging right so so I listed a lot of things that sometimes people list is on their wish list you know so called you know post human or transhumanists but most of those can be handled with physics and chemistry quite adequately I would I would not say that I would want to change my genome so that I could fly into space without a space suit but I what I listed I thought that are possible not addressed by physics and chemistry alone our longevity immunity and cognitive behavioral traits some of these will be addressed in adults through right conventional therapies there's there you can already see some conventional therapies that treat some of these you know caffeine metformin these sorts of things and there's and there's it's hard to raise an ethical issue when you're talking about small molecule drugs or probably if you have a safe and effective gene therapy applied to an adult human being and so I think that a lot of the red lines that we draw I'm not a big fan of drawing red lines ethical I think I'm a fan of having strong discussions and algorithms but it's much more complicated than a red line and the red lines that are drawn about you know worrying about embryo manipulation artists interacting us from real issues so if you manipulate an embryo let's say to be enhanced for cognitive it's gonna be 20 years before you see any impact of that on society while if you can change an adult cognitively that could spread in weeks through the internet everybody is doing do it yourself gene therapy on their brain I mean that sounds ludicrous but we live in a time of exponential change and I personally know several people that are already doing do-it-yourself gene therapy on themselves these are not wealthy individuals either I'm not encouraging you to do this okay but I'm just reporting that there are people are doing this if look at the latest MIT tech review for example okay let's take one or two more questions over here hi so I'm sure you're aware that the vast majority of diseases are actually epigenetic in nature and not genetic and so because epigenetics is so multifactorial it depends on things like stress in your diet and lifestyle should we really be using this has a genetic engineering has a crutch I'm instead of altering lifestyle or cleaning up the environment and things like that yeah so the you know the question is how many of these things can be fixed by life lifestyle and diet and tay-sachs to take an exam an extreme example is probably hard to fix with lifestyle and diet it is a genetic disease that is what pretty well understood and there's hundreds like it for diabetes on the other hand is something that is an epidemic and it is probably environmental and lifestyle and and it could be microbiome it might have a fix that has nothing to do with human genetics it has to do with environment include if you include your microbiome as part of your environment so yeah I think we should take a deep breath and make sure that we're not using you know bazooka when we could use a flyswatter but but not everything's gonna be that easy there's one way in the back there oh okay yes please yeah way in the back and then we'll come here and then we'll be we'll wrap it up yeah please in the back hey I know you said not to ask this but I'm still very curious what do you think are the advances required to actually make the next sleep in read and write to get the thousand or a million fold improvement I'm gonna ask you to just restate that so I make sure I get it right you know oh in at one point you're talking you're mentioning that you thought that we could still make a thousand or you know a million fold in our ability to read and write and I'm just curious what are the enabling technologies you think that are required to make that leap yeah so if you had asked me in 2003 or let's say at the beginning of that plot you know how we were gonna get a billion fold improvement I might have said very glibly miniaturization multiplexing and self-assembly but that's not a recipe if I had a recipe in the 1980s you know things would be different but roughly speaking I give you an example that we are actively pursuing is right now DNA synthesis is done by organic chemistry phosphoramidites it takes three minutes to add a base you can do many in parallel but it takes three minutes to do each of those in parallel while biochemistry polymerases can go up to a hundred thousand times faster than that that's not a recipe but it's a it's an indication of where one can go and and that and that speed turns into cost because the equipment whatever whatever the parallelism is however you're directing the pixels in an array the cost of that machine has to be amortized though before it's you know over a period of years and in the more cycles you can get per year the better so if that's that's a factor of 10 to the fifth for example for example yeah that's a question in the friend here please thank you I really enjoyed the talk and so machine learning and deep learning has really impacted biomedical research for example in medical imaging algorithms being able to identify cancers as effectively or more effectively than doctors I'm wondering in your opinion what sort of applications you see for deep learning machine learning even artificial intelligence in genetic engineering why so the question is about deep machine learning in biology in general and I think this simple hopefully not too glib answer is everything almost everything that we currently do could be augmented by machines I've you know I've gone on record as saying that human intelligence is has a lot of advantages over machines with we're that's a 20-watt computer while Watson the jeopardy winner machine is 85,000 watt brain but putting aside small human chauvinist comments like that I think that that the deep learning it has a huge promise in biology and there's going to be a little loop so for example we have a grant from the part of the brain initiative from AI ARPA where we're analyzing a cubic millimeter of visual cortex with amazing set of tools for calcium imaging and behaving a live behaving animal and getting the connection wiring diagram for that activity map to synapse level resolution in order to and this is the goal of the IRP project to help the machine learning community to handle complex visual tasks like self-driving cars viewing faces and security footage and so forth because these are not solve problems in their community and so so there would be a virtuous cycle where they help us read out the brain and then what we read out the brain can help them with their algorithms and through that two times and it'll be amazing well up it's probably a good place to stop but thank you very much to the audience for your for your your great questions and George I just like to say that I think you you're setting a wonderful example as a scientist who is not only thinking about the future and kind of bringing it about and going there going there with with appropriate caution but you're also coming out of the lab and you're talking about it publicly and I think that's we need more of that so I really applaud you for that and thank you for giving us a fantastic evening here Thank You Jennifer [Applause] [Music]

Channel: The Artificial Intelligence Channel

Views: 43,230

Rating: 4.8817968 out of 5

Keywords: George Church, CRISPR, synthetic biology, UC Berkeley, evolutionary biology, genome engineering

Id: Wswvf8Nrubg

Length: 80min 25sec (4825 seconds)

Published: Sat Jan 13 2018