I am a biohacker

David Ishee

Keoni Gandall

Opening remarks 1

There are refreshments in the back. There is beer for people who are of age. There's also canned wine. Again for people who are of age. Alright, alright. Zayner in the house. We have tons of refreshments for you guys. We have cold beer because scientists are socially awkward. Welcome to Biohack the Planet 2018. Velkom. We have people from Missippi. We have people from the Bay Area. Please have a seat. Please silence your cell phones.

We want to thank Founders Fund and Encryptgen and osfund for funding this year. A lot of biohacker grants were awarded this year, cash money. Also we have donated towards the Omni. All the chairs you are sitting on were purchased and cleaned and vacuumed. Without further adue, I'm going to introduce Josiah Zayner.

Opening remarks 2

Josiah Zayner

... Peter, Mycah and some ohter people. OBviously I couldn't put this thing on alone. I tried the first year but that failed. There's so many people who have hleped out with this. The Omni Commons of course. They let us use this space and it's really inexpensive. Counter Culture Labs is the bioahcker space in the back. This conference this year is a little bit different... if anoyone knows me, I hate doing the same things that everyone else does. I can't stand it. People give talks at conferences- we don't do that. What we're going to do is, we're going to do conversations throughout this whole conference. That means two people sitting up here with some scotch or tequila and just have a conversation. There will be a microphone up here and a wireless mic running around. Just ask questions that you want to ask-- you should be able to interact with all of these people. We will interact with them and with each other and hopefuly we'll have a good entertaining time. Hopefully, yeah.

Before we start, I wish I had some intro music. Any chance of that? Maybe some AC/DC. Anything would be great. Tomorrow the schedule is going to be-- there's some crazy stuff happening. In the last 3 days, we've had a few speakers cancel. We've survived. The schedule tomorrow will be a little bit wonky but it will all be amazing entertaining people that you will be able to listen to. It will all be great. Here we go.

I was trying to get you guys pumped up. It's not working. Esther was awesome. What we did is, we hid some prizes under the seats. Look under your seats, and if you find something here... if you find anything... A pipette. Oh my god. Wow, a pipette. You get a pipette, you get a pipette, you get a pipette. Thank you Ester, thank you for the hype. I really needed that. Anyway, we'll just start.

The first discussion we'll have is between two people you may not be familiar with. One is David Ishee and the other is Keoni Gandall. We have two people. We're going to have a discussion. These are pretty infamous people. Both of them have no college education and they are doing some synthetic biology, genetic engineering work. They can probably talk about themselves a lot better than I could. Please introduce yourselves and have a conversation.

Talk

DI: I am David Ishee. I am a dog breeder. I've spent the last 3 years... recently branching out to tweaking on myself. That's sort of me.

KG: Hello. I am Keoni Gandall. My goal is to make biotech more accessible to people. To make biotechnology something that anyone can work with or anyone can modify and build and create. Over the last year or so, I've been working with the Biobricks Foundation for free genes for people. Basically, my goal is to make it so that anyone fcan genetically modiufy any organism they want without any prior knowledge or without going through a lot of expensive formal training. Just making that accessible to everyone.

DI: You say that...

KG: The Biobricks Foundation is not associated with iGEM. It was founded by the same folks, some of which are here.

DI: So it's not iGEM?

KG: iGEM does a lot of things right but they do a few things wrong as well. They make it very expensive and the expense of going to iGEM is only going up not down. This makes it so that a lot of biohackers or biohacker labs or just people wanting to biohack themselves can't afford to go to iGEM. They are throwing that community under the rug.

DI: I looked at it once. It was like $5k to go to iGEM. So nevermind.

KG: Their parts registry is great. But not just iGEM should have access to that technology. Everyone should be able to have access to that DNA. They don't right now.

DI: ... just big institutions.

KG: I don't want that. Opposite world.

DI: You've been doing the thing with free genes, then. How is that been going?

KG: Yes. Good.

DI: Could you expand on that?

KG: It's been ramping up. Really getting good at making... something that I think everyone here is... automate yourself away, try to make it so that everything can be done by robots. You just setup a robot and then you can get your friend's protocols or your collaborators protocols. Everyone is sharing the same stuff, protocols, DNA, or robots.

DI: But that doesn't work.

KG: Some day it will. It's a small robot. Like the size of htis table.

DI: Oh, I could probably squeeze that in.

KG: do you want to make genetically engineered mamamls accessible to everyone?

DI: Oh, yeah. The first people I would want to work with are dog breeders. They have been genetically modifying dogs for 40,000 years. It's one of the oldest longest running genetic engineering experiment we have. We're using the same tools that we were using 40,000 years ago. We started doing this before agriculture and the tools haven't changed since then. There's room for improvement. You have over 600 different genetic diseases and some of them are just crazy perfect for CRISPR and things like that. Like in Dalmations-- three bases upstream of a PAM site is a single point mutation which is begging for cas9. The problem is that it's expensive, and it's tricky, and it's just not a skillset necessarily that dog breeders have. I want to show dog breeders that they can do it. And figure out a way that they could do it quickly.

KG: Could the same tech be used on humans?

DI: Actually I tried that. The same buffers kill human sperm. With a few tweaks, it could work.

Q: How did you make the human sperm?

DI: The old fashion way.

KG: How does that work in dogs? What's going on?

DI: Over the last couple of years... but it was really early. They get big, then get smaller, make milk, then stop making milk. I think this was because I was using luc the set.... several enzymes, ... I think that was the issue, so I swapped it with something simpler. But I've been optimizing my protocol, so now it's at 90% of the sperm get modified. So hopefully that's the most... We have 2 females pregnant right now, we'll know in October. Two females.

KG: Where can we find more information about that?

DI: Uh, facebook mostly. I should probably blog or something. You could just go to my website or find me on facebook. ((It's an uncool website he has.))

KG: What inspires you most?

DI: Let's see. Honestly, it's other people in the biohacking community. I see people doing crazy stuff. I got started because I saw someone do a TED talk. I see people in the community who are.... and then... other people are just-- there's just so much amazing stuff going on that people are getting some results. Some of this stuff is really advanced and really hard stuff. And people are putting in the time and the effort and making it work. That's what inspires me to keep drilling in. What about you?

KG: Mine is a little bit more abstract, not really personal. I'm really inspired by the computer revolution that happened in the 60s, 70s, 80s and 90s. The generation of computer software going from a model of like "we don't know what we're doing" still using vacuum tubes to the point where we have Linux and a whole operating syste mfor free that you can install on any laptop.

DI: Are we closer to vacuum tubes or closer to Linux? In biology?

KG: In biology, we are at vacuum tubes right now. We need to get to Linux. Right now you have two paths that aren't mutually exclusive-- a path driven by large corporate interests like pharma is really interested in synthetic biology for drug production, and we have another path which can be a grassroots method for indvidiauls interested in solving their problems in their local communities. Hopefully, right now, we're still on one path mainly where large corporations and big pharma gets ton of funding to do synthetic biology in their own biofoundries right next to MIT that aren't really sharing a lot of the stuff that they do. What I want to see happen is more of a grassroots movement where individuals are empowered to make a change in their local environments or communities, just like people in the original software revolution were able to do so.

DI: Right now... biohackers run into problems like nobody has any money, funding, equipment, reagents, everyone is struggling with barely getting what they need. You have a job, and then you spend nights and weekends. They don't have money. And a lot of this is open-source. The dog breeders are in a multi-billion dollar industry and they desperately need this expertise. Not just the tools but people that understand it. I think I could make a kit that would allow dog breeders with relatively little knowledge-- to give them the ability to do sperm-mediated gene transfer. It's a couple of extra steps on artificial insemination. If I could give them that, then they have the tools and the abilities to make the edits, but they don't have the understanding to design this. There's this whole other level of genetic design that I'm not expecting dog breeders to get into. The dog breeders and biohackers could together work cooperatively. If you're a dog breeder and you have bull dogs then they have a hip dysplasia and you want to fix that, so you go to someone in the biohcaking community who is a struggling postgrad with no money but they have the expertise to do the genetic design and you have the ability to dump the DNA in there... so working together, hte dog breeder can generate funds by selling the dogs and use that to pay the biohackers. I think these two communities can complement each other pretty well.

Q: What about the ethical struggle? It's so tricky. How is this going to be resolved?

DI: That's always the big thing. The ethics are always tricky when you talk about mammals. If you want to talk about yeast or bacteria, that's something else. You kill half and that's fine. But if you're talking about dogs, they can feel pain and people love them. It's different. I think for every dog breeder there's a challenge in at least the dog breeders that give a shit, there's something about producing healthy animals and animals that are working towards better and better in terms of health and performance and whatever you're breeding for. There's always a probability that you're going to get a dog that has a problem even with the healthiest lines. There's always some probabiliy. I had a buddy that had a dog born with one eye and no mouth. It didn't survive because it couldn't breath. It was just deformed. It does happen. If the edits you're making, especially as a first experiment, you have to-- the other side of it has to be something important-- some people use this for fashion where they want to make the pinkest bull dog. Honestly people would probably buy that. The problem is that if your end goal is to address a genetic disease, especially in something like Dalmations--- .... but basically, all purebred Dalmations have this problem-- there are dog breeds where the whole population has these genetic diseases. If your goal is to cure something like that which is otherwise uncurable and I understand then they could just crossbreed and I've been arguing that for 10 years. All my dog are crossbreed. But this just pushes it below the surface. To really fix it, on a population-wide level, you need to do more than just fix one individual and breed them because that tanks diversity. You need lots of dog breeders to do this.

Q: What would be the dog you would want to breed?

DI: The dogs that I have now are pretty good.

Q: No what skills, would it be able to fly? What's the dog you want?

DI: Let's say... it's 30 years from now and we have perfect control. Have you ever seen The Hobbit? There's these porgs and they ride them. There was one white one. It's gorgeous. I would like to ride it into work.

Q: Are you a dwarf?

DI: But yeah, not only does it give the breeders to do more than they have, but it also gives you the ability to.... So I think there will be a cost, but if it's used right, it will work out in the end.

Q: .... what are you working towards, what's your end goal?

KG: I want to build a cell from scratch. Ammonia is pretty good. I want to be able to take genes from an organism and put them together in less than a week for less than $1000 and get a full synthetic cell. It's much less personal than dogs, I admit. But I think it's cool that you can build life from scratch from the very essence and build it up exactly as you want it, lifeforms that have never been seen before or as you have imagined.

DI: Like porgs.

KG: Those have been imagined.

DI: ... nobody has imagined ...

Q: What would come from these cells? What organisms?

KG: I'm more interested in the journey of helping build those synthetic cells. But let people build whatever life they want from scratch.

DI: But won't that cause a zombie apocalypse?

KG: No. Are you going to turn the porgs into zombies? Hopefully not.

Q: Do you think we'll ever be to the point where you build multicellular organisms from scratch and if so, whta do you think about that? Have you only been thinking on the single-cell scale, or what about larger than that?

KG: I think it would be cool to have a Zerdling. Kra'aaw. But it's really a pokemon. I mean, it's completely realistic to believe that some day we will be able to build multicellular organisms from scratch.

DI: Pet dragons.

KG: Eventually we will get there. It will be so cool to be able to make pokemon from scratch.

Q: How far are we away from that?

KG: 100 years. 50 years. It depends on how the exponential goes. What part of the curve are we on?

DI: 30 years. I'm sure it's 30 years. If you start with multicellular organisms...

KG: If you look at the 70s or 80s, there's no flying cars.

DI: There are flying cars, they are just for rich people.

Q: They are actually called airplanes.

KG: Fair enough.

Q: Is the cell-from-scratch project your idea, and if so, is it based off of a step towards something? Is it like your childhood dreams of building an organism from scratch?

KG: I wanted to make a minimal plasmid. I figured out that was easy. But I wanted to do that with a virus next, and it turns out that viruses are already pretty minimized. In 9th grade, I said the next step is....

DI: What's your favorite virus?

KG: Ms2. 3500 base pairs. ssDNA into ecoli... 4 genes. It's the most amazing virus there is. ((applause)) I highly recommend looking at this virus on Wikipedia. ms2. It's all a secondary RNA structure. It's a crazy molecular machine. It produces 40,000 more of itself from a single cell. It's my favorite virus.

Q: You guys don't have like, zero background in biology or... went to university or anything. How did you guys self-start? What kind of groups or sources did you get? How did you start to get into genetic engineering?

DI: For me it was that I saw a TED talk and people were implying it was a lot easier. So I started googling stuff. There's actually shockingly large amounts of stuff on youtube. There's openwetware.org and the-odin.com and lots of kits and protocols from every company. If there's something weird that I don't know how to do, I just ask people and they tell me. Just try stuff online, fail a whole bunch of times, and the nfigure out the right thing to do.

KG: I got a biology book from a church bookfaire and got really interested in viruses. Only bacterial ones. In 7th grade, I was printing out genbank files of phage lambda stuff.... and I looked at a bacterial transformation kit in 7th grade.

DI: So the FBI has been watching you since about 7th grade?

KG: Hopefully.

Q: That means you're worth something.

KG: Exactly. How would you like to be remembered?

DI: Someone who did something meaningful. That's sort of a weird thing. You go to work, you go home, you go to work, you go home. And then you die. That's it. That's what you did. I work in the oil industry. My whole life product will be that I made some oil. It doesn't matter at all. If I had never lived, it wouldn't matter at all. I would rather help something to make something that lasts. If I can help all the dogs in the world to be healthier, even a little, then that impacts dogs. That's how I would rather be remembered.

Q: ...

Q: ... I remember last year, David you gave a talk and you got into the legislation that was coming down that would prevent you from doing what you want to do. So my question is, what kind of legislation has changed in this last year? How does that effect what you want to do? There have been changes.

DI: In 2017, the FDA stuff... as far as that, they haven't changed their policies. There's some interesting threats to take away their authority to handle animals and shift that to the USDA. I think tha twould be interesting because the USDA handles everything else with animals and agricultural animals. I think that would help. While you still would have ot prove it's okay to do transgenic stuff with dogs, that would be more effort, but to just take away diseases, USDA would be fine with that because it's just breeding. That's what I would like to see. If they do that, then that would be helpful. There was also a talk at the FDA about shifting more of its stance- it's more science backed and not sort of we're going to be really cautious and just grab everything and hope nothing weird happens.... but as far as the real rules, nothing has really changed. There's just been talk and rumor that things might change soon. I think things will change, I'm optimistic, which is dangerous when you're dealing with the government.

KG: I think Megan Palmer is out in the crowd. She is the person to talk to because she does lots of bioethics stuff like that ((boos from the audience)).

Q: I like the idea of the artificial cell. But what specifically what type of grassroots movements do you think should be setup?

KG: The technology that I'm talking about here is one that enables building a cell as a goal but how to get there is making it really easy to make a lot of DNA, verify it, and then make standards for that piece of DNA. Through doing that, you get a standardization of biology, you also get automated instructions and the costs of that going down. Finally, by making those two go down, the cost of monetary the amount of money to synthesize that gene and the cost of figuring out how to design it... you lower those two variables, then you make all bioengineering easier for everyone. Imagine if dog breeders could get that piece of DNA they need, for $10 instead of $1000. Imagine if any of the small breeding community even you could think of other animals or breweries or you could think of people producing bread or local products like yogurts... those communities now the ave the ability to use geneitc engineering to do whatever they need to do. Building a cell is the goal down the line, but what we have to do to aciheve that goal is what is going ot make the world a better place.

DI: So, it's like the moonshot that makes..

KG: Yes.

Q: ... How useful is it to go to school, anymore? Considering how fast the field moves.

DI: Since you work for a university, I'd like to hear your opinions on this.

KG: Oh boy. I work under Drew Endy. He's here today. It's going to be computer science in probably a decade or so. It's going to be how you perform what you've done is just determines what job you get or how useful that is or how useful you are or how you are received. It's not there right now, because biology is hard, and you can't do it until you get into grad school. If you don't have access to the technologies- and many people still don't-- then going to school can be useful to learn about that type of tech. But if you do have the access and you're passionate, then I think there are ways to perhaps strict schooling.

DI: I sure hope so. For me, I don't know what it's like on the other side. I never went to school. I dropped out in 5th grade. I don't know what they teach in universities. I've watched some lectures. I have found that there's just everything you would ever want to know on google. You could use Google Scholar and scihub to bypass the paywalls. You could see everything ever written. There's EdX. There's some really good sites. The only thing you don't have is hands-on, but if you build your own lab then you can do your own hands-on. Maybe it's slower, I don't know. I am glad there's some other route other than just getting an education and having 30 years to spare, or just having $100k. It's nice to see that there's more than one way. I don't know if there will be a point to universities in the near future. Maybe they will continue to be good for research. I think we're about to get to the point where it's not worth the money.

Q: ... if you were offered a position at MIT right now, would you say yes or no? The position in a PhD program right now, if they offered it to you right now, if you were paid $40k/year to get a PhD at MIT right now. Would you say yes or no?

KG: I would say no because I like the Bay Area.

DI: I would have to move to Boston? $35k/year there? I don't know. I would like to keep my quality of life. ((Would they fund the dog research?))

KG: I would probably finish what I'm doing now but then I would probably take it.

Q: I have a technical question. David, you do CRISPR?

DI: Not yet.

Q: There was a recent article a few months ago that claimed ....

DI: What I'm using is less precise than CRISPR. I'm just integrating GFP. I'm just verifying the process. The protocol I'm using has only been done in pigs and cows. I'm using random integration. It's way less precise than CRISPR. Most of the animals that have been produced by random integration of plasmids into genomes have been fine. So honestly, CRISPR is like 5% off-target effects or whatever... it's okay. But I also have a version of Cas9 that has been mutated to be high fidelity. So when I do the dalmation thing, the CRISPR-Cas9 that I will be using, the off-target mutation rate is less than random chance.

Q: ...

DI: Yes. Yes. The FDA will not be removing these dogs off my property. They will be here forever.

JZ: Any more questions? Last chance.

Q: I have a two part question. If you had the ability to do human genetic engineering, how would you go about doing it? What are the problems and possibilities of human genetic engineering?

KG: If I had to do human genetic engineering, the first thing I'd do is probably, is get The Odin's kit to genetically modify frogs, and test there first, and then move that system to myself. I'm not sure if I would be comfortbale genetically modifying my kids quite yet. I don't want to hurt them. In the future, we'll be able to remove most diseases that are genetic from the human line. I think that would be fantastic.

DI: A little more interesting.... almost everybody gets behind the idea of fixing hereditary diseases. What are your thoughts about going beyond bringing them back to healthy?

KG: It's controversial, but...

DI: Let's say it's proven to enhance their whatever.

KG: I am fine with it. I think advancing humanity is good and it will benefit everyone. In the early day of medicines, it starts off super expensive and only the wealthy can get access. But the price gets lower and lower. The tech gets better and there will be grassroots movements. What I hope to see is that, let's say there's an improvement ot the human condition. I would want that eventually available to everyone and there should be steps taken right now to make sure that future is available.

DI: So super humans are okay, as long as everyone can be super humans.

KG: Yes.

JZ: Last question.

Q: Something about biobrick foundation.... and I know you're interested in... synthetic biology... how are you going to help that from inside the organization to help diybio and synthetic biology cheaper?

KG: Free genes. We have a free genes project where people can submit genes they need, and we will synthesize them for free. That's a pretty low price. Further than that, the software that we're building to be able to do that, and the robots that we're managing to be able to do that, we're open-sourcing all of that. Anyone can create these, and get thousands of genes. Once you lower that cost and sometimes, centralize it, but anyone can do it- you lower the cost of actually building genes. So you're building software in an organization that will be able to lower the cost for you to build DNA.

Q: What about economic goals or challenges you have faced when trying to drive down this cost?

KG: We can talk later. The main challenge that we face.. are technical ones. It turns out, how you manage all of this data, how you sequence verify 2000 genes. It's hard. How you manage all those robots- what about 3 robots running at once running on the same plates or same pieces of DNA? Figuring all of that has been a challenge. We're beginning to solve it. It turns out that probably once we finish this, in software, we can give it to you and then you don't have to figure out those problems.