path: root/transcripts/pcsbi-2010/2010-07-08

There are components in the cell, uh, George Church and others are going to be using those components, and we think we will as well. They have cell-free systems that can be reconstructed. It's interesting science, but it's irrelevant to these arguments for booting up a new piece of chemical software. One of the most important experiments was in 2007 where we isolated the DNA and chromosome from one species, transplated it into another cell, replacing the DNA in that cell. And it converted the cell into the species that we isolated the DNA from. So it's like putting different DNA into you, and converting you into an other species. We can do that at the single cell level. Doing it at the multiple cell level is uh perhaps decades off, but not centuries away. So these are important concepts about starting with digital code and DNA. It is software. It does build its own hardware. That's an interesting science question: what do we need? We need some ribosomes, a few tRNAs, some other things, some lipids, can we get some cells booted up from that? I think that's going to be important about understanding origins of life. But we are in fact building upon 3.5 billion years of evolution. Um. The arguments that because we're using genes that we discovered, versus inventing new genes, it's kind of spurious, like saying Tesla did not build a new electric car because they bought the batteries from one source, and they bought the electric motor from another source, they combined that to make a Tesla. Um, which is a pretty exciting electric car. My team has discovered the majority of genes known to science. We're up to 40M, there were less than 1M when we started. These are going to be the future design components. Biobricks are important teaching tools. It's great for getting students involved, but the number of genes will top out over the number of 200 or 300 million. We're dealing with a lot of design components. Nobody is going to patent them, and combining them in new ways, now using these tools that we had, the proof of concept experiment, that's the future of this field.

We couldn't do any of this until we did this study that was just published in Science. We're able to make these really large pieces of DNA, but until you can boot them up in a cell, it was just an interesting academic exercise. So, it's very different from what has happened before in molecular biology. This is a new set of tools, starting from a new vantage point. It changes a lot of the rules. Scientists sort of controlled uh who got what, whether they sent them their cell line or DNA clone for their gene. Now, anybody who has access to the internet, that information is in the public databases, you can download and make those genes, now any virus sequence that's in the public databases, can be pretty readily re-made, fortunately not all of them are the DNA-effective. Smallpox is one of them, where just having the DNA from smallpox on its own can't just boot up readily. But these tools are there, and it's a different starting point. All you need is the general information and a DNA synthesizer.

So, building the pieces of DNA was an interesting technological challenge. Getting it booted it up was straight-forward biology and molecular biology. None of it is cloning. So these are totally misuses.. cloning means anything and everything to biologists that sort of collects terms. It's making copies of cell, copies of DNA, splicing DNA, so, we think it's an irrelevant term for what we do. We used the term synthetic cell, because every protein in the cell all the constructions in the cell are derived from the synthetic DNA.

The cell that we used as the recipient cell, all its characteristics are 100% gone after a few replications, so everything in the cell that we have is from that synthetic DNA and therefore we defined it as a synthetic cell. It's a cell that never existed before, uh, of course we used copies of existing genomes. I agree with the very statements- that we're very early on in our knowledge of biology. But we definitely have new tools now to get there. We're using somebody's tools to make new vaccines, so we have a program that is funded by the NIH, to make synthetic components of every flu vaccine that we and others have sequenced, and we can combine these and make a new flu vaccine candidate in less than 24 hours. We're working with Norvartis (?), and it's possible that the flu vaccine that you get next year will be from these synthetic DNA and synthetic genomic technologies.

It was announced last year that we have a program with Exxon Mobile to get cells to capture CO2 and make uh basically a biocrude that can go into refineries. We have not found any cells that can do this naturally at the levels that are required. At the very minimum it is going to need extensive engineering. I am absolutely certain that by the time we get to version 2.0 of these cells they will be completely synthetic as will most things going forward in an industrial environment. Definitions are important, the definitions can be found in our scientific publications. I think this is an area that Drew Endy's students show are more limited by our imagination.

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Back to our early recommendations on licensing being, .. ecosystem of synthetic biology, is important, we need to have surveillance and testing of systems that are proposed to go in. This is just not restricted to bacteria. We have a very active synthetic biology community and human DIY community. Some of my undergraduates have gone and sequenced parts of their genome on their own without FDA approval and without special equipment. This is a whole-nother subject that we are not going to talk about, DIY, do it ourself, do it yourself, bioweathermap and so on. We've studied vaccination, that's another topic for another day, genomic engineering, some success stories, Artimesomeeon, Dupont .. million dollar project, very successful, 90% of the theoretical yield only inolved 8 foreign gene plus 13 up and .. Methylelososne from .. Methyleleosene. 27 changes was a lot of work back then. I am going to talk about 100s of changes that we've incorporated. These are two other companies that I helped start, that are.. not in the future, but are already making 1000s of liters of production-scale fuels either from biomass or carbon dioxide and light. These are making alkanes, diesel and gasoline. The success of comparative genomics, you can look at algae and .. to find those genes, you can take, look at .. and over.. produce them. 

Rob Carlson elluded to this, .. this exponential curve from 1.5 to 10 fold. More importantly this is a gap between our recent huge increase due to the second generation sequencing and synthesis. We're still stuck in the first generation for gene synthesis and those four companies, and genome synthesis. We're using first generation sequencing and synthesis for the most part. There are 21 next generation sequencing technologies and 21 companies that go with it, and I am an advisor for about 16 of them. There's also next generation synthesis off of chips, since around 2004, this has lagged a little bit behind for making genes and genomes, but it's certainly terrific for making very short constructs.

* A conformational switch controls the DNA cleavage activity of lambda integrase
* Design, activity and structure of a highly specific artificial endonuclease

Getting those, and working in cells; it's one thing to make DNA, getting it to work in cells; there are protein-based specificity tools, and more general tools which are DNA based and homology based, they do not require particular tools to put it in precise locations in the genome. Some of these require ssDNA, we've automated this in order to bring down the cost. This is multi-plexed automated genome engineering (MAGE) this has one particular implementation shown on this slide, it's a catch-all phrase, this one uses single stranded oligonucleotides, that use CAD to optimize secondary structure and to optimize the position and length, um, you have to have mismatch repair turned off for some of these, and there are special proteins, but the key point is that in a few years we moved from an efficiency of 1e-4 (1 in 10,000) to 25%, 100%, and now we can get up to 8 mutations per 2 hour cycle and we can just continue to cycle 8 changes precisely in the genome where-ever you want. You can make up to a billion different changes in a population. I'll show you an example of where we did 100,000. This is Harrison's prototype, CAD of the upgrade, this is the actual upgrade, this is applying it where we made 100,000 genomes (not one by one but in a mixture), but it shows the awesome power of accelerated evolution in a laboratory where we can make these 100,000 genomes focusing all of the changes in the known pathways including putting in some genes from other organisms, and in three days we can get the highest yields that we have ever seen for this hydrocarbon lipipine, which makes tomatoes red. It evolved (involved?) the order of 24 genes. 

Another project that we have done which is less combinatorial, less evolutionary-based, where we wanted to make a precise gneome more resistant to particular viruses, and allowing nucleotides to incorporate efficiently. Here we changed all of the codons for examples of codons (TAG) into TAA (?? i got this wrong) in order to free up that codon and allow us to delete the cellular factor that recognizes it.

SynBERC - Synthetic Biology Engineering Research Center

This can be generalized, there are 64 codons of these triplets, and we've targeted nine of them. This allows us to do three things: multi-virus resistance, safety features,  

There are 64 codons of these tirplets, we've targeted nine of them. This allows us to do three things: new amino acids, safety features, and multivirus resistance which itself is a safety feature. We've done one of these nine codons that we are targetting out of 64, we've synthesized all of the DNA to do the remaining eight, at least a proof of concept on the essential genes, and there's another topic- another project- where we are making new ribosomes which is an in vitro system which has interesting commercial applications. Just changing these nine codons would require 2.7% of the genome, but if we're making these 90-mers, we have to tile the genome 2 and a half fold over, we're essentially changing the genome, even though it's just 2.7% of it. Doing it more efficient, all synthesis all at once, where we will probably have multiple failures, doing it one at a time.

Just as a last slide or two, is this issue of safety in terms of isolation. You can have physical isolation, biological isolation. The changing of a genetic code gives you biological isolation- genes can neither go in or out that are functioning. Critics of GM organisms..

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New characteristics. That's part of synthetic biology that has been different from genetic engineering. It's not entirely distinct from systems biology- math, the ability to predict and design what we do. DNA synthesis is an enabling technology. You'll see from the first few slides, if we're talking about biology as engineering and getting easier, Drew Endy was talking about the tech gap between writing DNA and knowing what to write. Under the umbrella of synthetic iology, there's DNA synthesis using it at a mirnimal level. In that vain, this goal of engineering biology, what's our biological workspace? Microbes being a substrate of choice, because of its relative simplicity? It's less complex than a mammalian cell, and also plants, and from an applications perspective, if these are the biological substrates that we want to work with, the applications become pretty clear: therapeutics including pharmaceuticals, small-scale pharma, biologics, protein-therapeutics or more complex therapeutic agents, fuels and energy, not restricted to fuels, chemicals which may be part of the pharmaceuticals, new ways for materials to have renewable materials, get rid of the polyproplyene bottles, agriculture as we go from plants, the biological progression from plants, the potential to expand even what we've seen before with GMO for agricultural.

Controlling transgene expression in subctaneous implants using a skin lotion containing the apple metabolite phloretin

This has been funding, 42.5M from the Gates foundation, and Amyris, and the Keasling lab to develop this technology. One of the interesting things is that the University of California has to agree jadto agree to make the licenses basically free, and the commitment would be that they would develop this at-cost production, this would be a ... Amyris has transitioned this, it's in the hands of industrial manufacturing. They have switched their focus to fuels, so it's an example of how the basic technology of these achievements, synthetic biology and 

Non-fermattive pathways for synthesis of branched-chain higher alcohols as biofuels

.. as it applies to fuels. We've talked about microbes, there are efforts and achievements in synthetic biology that are going into more complex systems. This is a paper from Martin Fueznegger groups, from PNAS a year ago, about a circuit for controlling gene expression for implants. So the idea is that they were able to take pieces from bacterial cells in order to put together a regulatory element that would respond to an element, that would respond to a skin lotion, they could have subcutaneous implants, and if you applied this lotion, you would get gene expression. Again this notion of a circuit where you can control expression of a gene from the introduction of a small molecule. Activating gene expression with perhaps novel forms of gene therapy, subcutaneous implant, not modifying the genome with more I would say complex perspectives of gene therapy where you're looking at examples of removing stem cells, re-engineering them, putting them back in, this would be an implant that would be distinct from the native or human chromosome.

LS9, Inc.

Biologically templated photocatlytic nanostructures for sustained light-driven water oxidation
Synthetic protein scaffolds provide modular control over metabolic flux.
A synthetic oscillatory netwrok of transcriptional regulation.
A synthetic gene-metabolic oscillator
A tunable synthetic mammalian oscillator
A synchronized quorum of genetic clocks
Creation of a Bacterial Cell Controlled by a Chemically Synthesized Gneome

Moving on to fuels, I've already mentioned Amyris, this is from the University of California Los Angeles published in Nature. This has been licensed by Gevo. Scale, optimzation and getting something really industrial viable. This is a case where this work was licensed by a company and they are actively working to commercialize this process, similar to the work done with.

.. This is just a screenshot from their website. They really do talk about themselves as being a synthetic biology company, being able to take advantage of all of the extensive information from genome sequencing projects, but increasingly the tools and technologies that we are developing for synthesis and construction of biology. They are relying and focusing on fuels, but they are also focusing on biochemicals.

This is an example of what I mentioned before: energy but not being fuels related. This was a paper released in April of this year from a lab at MIT where again because of the multiple definitions of synthetic biology.. They used M13 phage as a templating device, so they were able to use them to form self-assembled structures, and the phage actually integrated with the inorganic often metals, and they are able to form higher-order structures. This is a case of using biological inspiration, or using biology as a template to make these nanostructures. But we can certainly think about how to expand this about having the power of synthetic biology and constructive biology, to be able to re-design these phages in different ways, so that the types of structures that we get are a little more complex.

One particular application was putting together a strructure that allows you to get photosynthesis. So they had light-driven water splitting, with the idea of using this for energy storage because you could capture now the hydrogen that comes from the splitting of water, and that the hydrogen could be stored and used at a later time. Whereas if we think about traditional solar energy, you have it available when the sun is out, and not available when the sun is not out.

This is from my lab, in collaboration with Keasling. They have the ability to make a pathway for gluceric cid...

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No, thank you very much for that list, and also for ending with the challenge that we have ahead. Keeping with the format that we have used before, I ask the commissioners to get their thoughts together, but I would return the favor and if anyone would like to fofer the first question.

Thank you all. Let me begin with a question to Craig, if I may. Uh, the potential power of synthetic biology creates hopes and it creates fears, and we're all too well aware now of the fears and I want to begin with the fears, as well. So, you mentioned a one-day production of vaccines for a flu, for example. Here's my question to you. What is the single hope that we should most believe in from synthetic biology moving forward? And, it would only be in incumbetent of me, to ask the same question as regard to fear. What is the single fear that we should take most seriously?

Venter: They both give me wide latitude, so I appreciate that. Don't make it too wide. On the hope side, obviously our own teams and others are trying to do as well, we need new tools to make new medicines a lot faster. Particularly with vaccines. It took quite a while with H1N1 the proper response, in part because the rate of building and deciding on seed stocks and in part because we're using 100 year old technology with chicken eggs to produce vaccines. Both need to change and quickly, but with rapid sequencing and all of these changes in reading the genetic code, and now the ability to quickly write the genetic code, it's now hours instead of weeks and months to make new seed stocks. The potential applications, because we can design cells with hundreds to thousands of energetic variation.. diseases like HIV that we're chatting about with that change in their genetic code very rapidly. Things like rhinovirus, we don't have a vaccine against the common cold because the virus evolves very rapidly. Designing things with the same rate of evolution or covering the spectrum of energetic variations gives us whole new ways to approach vaccines that never existed before. On the environmental side, I think it's clear that we need to do something different as we go from 6.5 to 9 to 10 billion people. We can't keep doing what we're doing. All of these different attempts need to be successful in creating new sources of fuel and energy and food or humanity will be irreversibly damaged and altered. We're a society dependent on science now, for our future. Biology is a key part of that future science. Synthetic biology, synthetic genomics are key, I hope, components of altering that future. On the fear side, the worst scenario is what happened in computing because we're talking about software: people make computer viruses that cause a lot of economic damage, but we don't want the same mentality going into making new animal or plant viruses either inadvertently or purposefully, and some of that can be readily prevented by straight-forward regulations, but obviously nobody develops new tech ever produce harm to others, we just would like to see the benefits. I think the molecular biology community has a pretty good track record from the last several decades because of the guidelines and rules that we've all been working under.

I also want to direct this question to Dr. Venter. Um, I've heard both you use and the literature, people have talked about the publication of science as a proof of concept. What is it exactly that it is proving? In part, because as I understand it, the cell wall of the bacteria was used in the first generation, and it's a natural organism that has been synthesized, what is it that it proves, how significant is that proof of concept, but second, building on that, looking forward, you might be working on algae or other multi-cellular organism where the genetic information is in the nucleus of the cell, rather than as a single strand. How far away are we from that? is that the proof of concept that would propel the field forward?

So, what's been possible in molecular biology is what several people described this morning as changing one or two things in a cell, and inserting that into plasmids. While we have evidence in evolution that many bacteria evolved by taking up entire chromosomes, and there are two chromosomes from two very different.. very clearly different origins, so they probably hapepned through these processes. But never before have we molecular biologists been able to take an entire bacterial chromosome, an entire chromosome of anything more than a small virus, transpl,ant it into a cell of one type and convert that cell into another type. Then you add to that, the digital code in the cfomputer, making the entire chromosome from scratch. Means now that we have the means to start with that digital code, make dramatic changes, while we have the basis of an existing organism, we made substantial changes to it, we inserted the names of 46 authors, several quotations, it's the first genome with its first built-in website, web address, these may seem like trivial changes, but uh they clearly identify it as a synthetically made chromosome, something we think is critical for this field and activated that, and completely transformed that one cell into a new cell. It was not just trivial.. one base pair set us back 3mo, one error in 3M bp did not enable this to happen. So, it's now because it's a proof of concept, we do know how to do it, and now we can make much more extensive modifications. We're building a robot to do combinatorial synthesis, where instead of making one chromosome over ten years, our goal is to make a million or so per day by randomly sorting genes or selecting the very specific ones and selecting for living cells that you can't get.. it's not a species that existed before. It's very similar to a pre-existing cell, but it grows substantially faster because of the 14 genes that we elliminated. "And on the multicellular front?" So that we don't get the negative consequences or the unintended ones of that..

"George, you've written on this way or not." I don't know if this is an ethical or policy issue, but many of the previous discussions, the conclusions have been that we're having more discussions. I actually think this is a place where we can do more than this, we can focus on licensing and surveillance, I don't know if this is ethics or not. What has hapepned since 1999 si that this exponential curve has gotten steeper. I would say that it's time to go beyond more discussions. 

Following on that, it seems that we have heard a lot from the previous panel and the three of you, about the wide-spread availability, the ability to do it in your garage, that seems to be about obtaining the sequences and the synthesis. And that generates worries, and my question is how big a step is it from having the sequence to actually getting it to work in a biological system, and is that gap big enough that we shouldn't be as fearful of this being misused, or the possibility of regulation or safeguards at that step that would be helpful? True or false?

That wasn't a yes no question, sorry. Then I would say, false. I think that with each new cellular system, and by the way, the Mycoplasma do not have a cell wall, and that's why we chose them - they had a simple plasma membrane which made it simple to make the DNA get across. At the JCVI, we're using spheroplasts that had a plasma membrane. We are at the earliest stages, we need to figure out how these tools are. Getting DNA past cell walls are tough. The two areas go in parallel. The design and synthesis and booting it up. Our worry was that we were going to have a large macromolecu.le, the largest man-made structure ever, and we couldn't activate it, because of one single error. It would have to be optimized for each individual biological systems. It's totally different with cell walls, without cell walls, plants and bacteria. This is going to be a very rapidly expanding research- regulation will be hard. Guidelines at the ... tough.. there's other ways to get around things.

This is going to be a very rapidly expanding area of research and probably difficult to regulate, the guidelines that get set up for approving projects at the institutional level with broader-guidelines at the funding level, and even though our work was not federally funded because our institution is a major federal grant recipient, we have to follow the rules regardless of the funded project that the government chooses. This has been a wonderful example of how to proceed, expanding the repetoire and expanding the way we go on with this.

I might be misinterpreting the question. The information is free. You can go to the NCBI database and get as much sequence as you want. It is cheaper, but still not trivial to pay for synthesis. My lab does not yet, we do not synthesis everything. We do a tremendous amount of PCR, I just had a meeting with a student a day ago, we can get these things synthesized, because it's $3k, but you can't get the 12 other variants that you want, because it's $36k. Some of it is access. Certainly we are not at $10 per base, but we're still under a $1/base, and we're not at a dime per base. At the levels of small amounts of orders, you can get negotations to get 10 cents to 25 cents a base if you want a lot of sequence. I might be misunderstanding your question. Whether or not you're talking about institutional access versus not, skilled labor versus not. Access in terms of cost, the things that may invoke a lot of fear and apprehension are beyond the cost of most non-institutional players, and then because we're talking about difficult biology, and one base pair setting you back three months, there's still a lot to do based on a skilled or unskilled.

Just to follow-up, I appreciate hearing that it is a little more difficult than just doing it in your garage to get the sequencing done, but I was talking about the next step, and whether or not people could do it in their garage, to get it to replicate in their cells, and whether or not that is an impoprtant place that regulation safeguards should be placed to make sure it doesn't get into the wrong hands.

if you read the blogs, everyone wants things to glow with green fluorescent protein. It's relatively easily to order a gene, order a plasmid that has a promoter and a terminator on one end, and transform that in your garage and say hey, it glows. It's very difficult to get your dog to glow. There's a level of complexity there, getting one gene to work in a garage with a junior high school is pretty close to trivial. The types of things that Venter and I did, are not going to happen in the garage with 14 year-olds. It's very expensive. 

Thank you very much for your presentations. You talked about some need for regulation, and I was wondering if you could comment on whether all of these things that would fall under the definition of synthetic biology are covered under existing regulations, because of certainly we have regulations on how to handle anthrax, ebola, or do you feel that we need to have new and different types of regulations to deal with issues in synthetic biology.

We certainly have recombinant DNA regulations, but most of this is dependent on federal grants, or in some other way being a responsible citizen. What we don't really have is surveillance that the regulations are being obeyed, by all citizens, not just the standard members of society. WE do not have many regulations on safety testing, even for things that might get into the environment. We take safety testing for granted, there's relatively little of that in biology. There are some gaps that we need to pay attention to. 

There are no limitations on what you can order from an oligonucleotide synthesis company. They are not required to list or screen against volatile agents. DNA synthesis is a global effort. If you can't get what you want here, you can order it in Germany or India or China, you can buy DNA synthesizers off of ebay, so maybe there are, people earlier, four companies that are probably 90% of the synthesis in the US even though they are not all in the US, getting them to require a screening against, ordering institution that could go towards the frivulous use. There is a lot of home-brew biology being done in kitchens, it's a new trend, Drew Endy has been in the past, it's great to see, he's stopped encouraging biohacking. We want some reasonable restraints on that, without destroying the wonderful creativity that these kids are doing, to come up with new circuitry that the kids can come up with. I don't think it's covered with existing regulations.

It would be possible to take 100-mers, to recognize that they may come from a pathogenic organism, order a bunch from one company, and be able to put them together. At the level of 100-mers, and anything over an 18-mer, you could get a pretty good trend of what someone is trying to do. The signatures are pretty clear-cut. Also these companies are beginning to coordinate voluntarily, this would be nice to be backed up with regulation. They are voluntarily doing this- splitting an order over four companies would be an alarming event, which combining with sequences that could be recognized, you could put the story together. There will be on-going efforts to get around this.

Entirely based on computational algorithsm, and testing, you could put that story together in hours. Government agencies that are willing to act in hours, that would be great.

I think you've gotten to the heart of a lot of my concerns, as you know. Asymitrio Cult, had an irreavalent strain of anthra, however that is a low hurdle to overcome in the ongoing biological processes, so I've heard a couple of different things. This is still difficult to do, very difficult, but the second part is that it is getting easier. I think that Rahj brought up an interesting point. There are regulations in place now, for traditional biology, being able to reproduce organisms that are on the toxins list or select agents list. But we are talking more about biobricks now, which are quite frankly not part of the regulation for BSAT, so what concerns, and I think you've expressed them, this evolving technology, and getting around the BSAT, the security measures that we would need to take to make sure these would not happen, and what is the balance, you've been prevvied of the latest BSAT, in balancing scientific discovery versus the security for the american people.

I don't actually think that this is a trade-off between security and science discovery. If this is properly implemented, where most of the effort is in developing computer software and getting compliance at the company level and surveillance at the government level. The researchers shouldn't even see it, and they could get on their work. If you require them to sign a piece of paper every few minutes, every time they pipette something, I think that's unlikely. I think some serious computational efforts are in order.

I'm interested in the money behind it all. It's a very expensive proposition to come up with a new cell as you did with over 40M dollars, or new products, and mostly funded now by often small biotech companies, with venture capital kind of money. Are there recommendations for encouraging the entrepreneurship, or not having so tight control or getting a payback on the amount of money you put into a project? Getting that payback quickly, and encouraging entrepreneurs to work on projects. There are huge profits in this if this works out in biofuels or energy, if it continues. Do you have thoughts or recommendations?

In my opinion, this current system is quite healthy to the contrast the one that Rob Carlson described. Most of my experience with dozens of companies, they can get the job done without spending tons of money on lawyers. You don't need the patents, you need the know-how. I have very few examples of patents getting in the way of academic research, and generally not getting in the way of startups. This is such a vibrant field, people are inventing so quickly, they invent around patents, or don't even concern themselves about patents. Going from small to large is quite quickly. LS9 and Exxron, they have Proctor and Gamble and Shevron, this is all in a short number of years. This is healthy.

It's healthy and critical- to get an ecological benefit of taking carbon out of the ground and putting it into the atmosphere, we need that to work out. There have been some recent changes in the stock market; stepped in where the government hasn't. Genentech and private investment, the majority comes from private investment. In our case, we would have been stuck in 2003 with a small synthetic virus if we did not have independent money to start synthetic genomics. 

I would only add to that, I agree with what's been said, I think one of the impediments to progress if you will, that can arise if the all of the achievements are done individually, is one of the very big goals of synthetic biology is to have standardization and interoperability. one of the ways that the federal government can help with this is to promote something in the community to organize on a reuglar basis around what should those standards should be, so that you don't have innovation happening in isolation, so that we have technologies evolving independently, and netwroking those becomes difficult. As we dream about synthetic biology, and ...


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Pubsliehd a paper in Science, it's great for understanding the concepts, converting it into the reality, where you can buy the fuel at the gas pump, that could only work in an economically competitive environment. All of these companies like LS9 will only survive if they have an economically competitive product. Any new fuels have to be available and they have to be cheaper than existing fuels, or at least cost-competitive. We need economic driving forces to pull this stuff much more rapidly. I don't see limitation of access, we need access pretty rapidly to CO2-based fuels as an example.

Before I go to NIDA and Anitya, I had a thought that was dropped. In connecting to something that you said. It's the flip side of regulation. Stimulation. one of the ways to insure safety, and in a gressykq-eseuqe way, have the ability to skate ahead of the puck, were you suggesting, or leave it open, the question that we had sort of posed in the prior panel, what would be the very next thing to be funded? Where would we like to see, in view of having a knowledge base and an ability to advance the kinds, and have the deeper knowledge to help ensure, that we can recognize, as Dr. Church mentioned, some of the sinister or potentially sinister application of these things? What would you fund, standardization?

It's a difficult question. So, I think about for example the BIOFAB which Drew Endy is funding and directing. It has an ambitious goal of being a focal point where you can develop parts to use a term in synthetic biology, some discrete pieces of DNA that encode for biological function, characterize them, composability, what happens when you put this thing with another thing, and it's an ambitious goal, and it's great, and they have $2M in 2 years, what happens when that is gone? There is a need for an effort that is more ambitious in scope, much bigger in scope, let's bring, that does two things: (1) serve as a forum or fora for bringing different players together and brainstorming and saying, here's what we're doing, ehre's what this person is doing, how do we interface this for setting a standard, so that new tech will fit in very well. Then how do we set priorities and safely, and so, I'll jumep ahead to a thought- the Synberc experience, we had discussions all along, we had a .. human practices, biosafety and biosecurity and intellectual property and silo applications. So, but that's a small number of people, and an increasingly populated academic field. If you look at the number of people associating with synthetic biology, it's grown astronomically over the past few years. If you take NSF's investment into synberc, and DOE's investment in the joint bio.. institute.. It's real money, it's not trivial, but it's a very small number of people that it impacts. But I would like to see efforts that are designed to bring the community together, and so that we could be more progressive and as opposed to re-active. We're in the midst now where we go with this, here's how it's all wrong, and I'm not saying it's not going to get us where we're going, if we want to bridge the technological divide, where we have the potentials and exciting technologies that have a real impact on humanity, we need to bring this forward quickly.

The two questions that I get most often, people are worried about bioterrorism, and environmental release. It depends on where you are, which one is first or second. George gave some wonderful examples of safety mechanisms that would be built in. It would be nice to have orders of magnitude more, suicide genes, large algae plants, modified organisms, they need to not be able to survive in the environment, suicide genes, artificial amino acids, chemical toxins, expanding the reepertorie of what safe and secure means, that would be a very beneficial thing.

Just quickly- I think it's coupling this question about small versusl arge. Large has some certain safety advantages, when we get to large manufacturing of automobiles did we start to get high levels of safety, as technology gets to a certain point, amateurs stop making it. I made a computer when I was young, the know-how starts to fade away, which is a mixed blessing, but from a safety standpoint, it's important.

It made me very proud of my heritage, engineering. One of the things that has been demonstrated in physical systems is that it is far more effective to design-in safety rather than regulate safety. Anita is next. 

>> Thank you. I really have appreciated all these remarks. I wanted to ask Dr. Prather a very specific question about something that you said toward the end of your slides. You had a slide in which you made the intriguing point our synthesis capabilities exceed our design capabilities. We know how, but not what. Can you elaborate on this? The reason I want you to elaborate, we know how but not what, does that point to limitations of the applications that might be forth coming from this science?

It absolutely does.

http://www.tvworldwide.com/events/bioethics/100708/globe_show/default_go.cfm?live=1&type=flv


starting with digital code and starting with DNA. It is software and it does build its own hardware. I think that's an interesting science question. What do we need? Do we need some ribosomes and TRNAs and lipids and can we get some cells bootup. I think that's going to be important about understanding origins of life. But we are in fact building upon 3.5 billion years of evolution. The argument is because we're using genes that we have discovered versus inventing a new gene is a spurious one. It's like they didn't create a new car because they built a battery from one source and they combined that to make the Tesla, a pretty exciting electric car. My team has discovered a variety of genes known to science. We're up to 40 million. There were less than 1 million when we started. These are going to be the future design components.
I think biobricks are an important teaching tool. It's great for getting students involved but the number of genes on this planet I'm sure will top out somewhere over 200 or 300 million. We're dealing with a lot of design components. Nobody is going to patent them. And I think combining those in new ways, now using these tools we had in the proof of context experiment is what the future of this field is going to be. We couldn't do any of this until we did the study that was just published in science. We were able to make these really large pieces of DNA. But until you can boot them up in a cell and get them activated, it was an interesting academic exercise. So it's very different from what's happened before in molecular biology. This is a new set of tools starting from a new vantage point. And as Drew said, it changes a lot of the rules. Scientists sort of controlled who got what, whether they sent them their cell line or their DNA clone for a gene. Now anybody who has access to the Internet, if that information is in the public databases, you can download it and you can make those genes. Now, any virus sequence that's in the public databases can be pretty readily remade. Fortunately, not all of them is the DNA ineffective. Smallpox is one of them where having the DNA from smallpox on its own can't just boot up readily. But these tools are there. It's a different startling point -- starting point. All you need is the digital information in a DNA synthesizer. So building the pieces of DNA was an interesting technological challenge. Getting booted up was straightforward biology and molecular biology. None of it is cloning. These are totally misuses of terms. Cloning means everything and anything to biologiesists and it's sort of a collecting term and making copies of cells. It's making copies of DNA. It's splicing DNA. So we think it's an irrelevant term for what we do. We use the term synthetic cell because every protein in the cell, all the constructions in the cell are derived from the synthetic DNA. The cell that we use as the recipient cell, all its characteristics are 100% gone after a few replications. So everything in the cell that we have is from that synthetic DNA and, therefore, we define it as a synthetic cell. It's a cell that never existed before. Of course, we use copies of existing genomes. I agree with the very statements we're very early on in our knowledge of biology, but we definitely have new tools now to get there. We're using some of these tools to make new vaccines so we have a program that is funded by the NIH to make synthetic components of every flu vaccine that we and others have ever sequenced. And we can recombine these and make a new flu vaccine candidate in less than 24 hours that we're working with no virus and it's very possible the flu vaccine you get next year will be from these synthetic DNA, synthetic genomic technologies. It was announced last year that we have a program with ExxonMobil to try and get cells to capture CO2 and make basically a biocrude that can go into refineries. We still have not found any cells that can do this naturally at the levels that are required. So at the very minimum, it's going to need extensive engineering, but I'm absolutely certain, at least by the time we get to version 2.0 of the cells, they will be completely synthetic as will most things going forward in an industrial environment. Definitions are important. The definitions can be found in our scientific publication. I think this is an area that Drew Endy's students show we are limited more by our imaginations now than any technological limitations. I think having an intelligent ethical framework for this new science to emerge in is absolutely critical. Thank you very much.
>> Thank you. We appreciate your views and clarification on the definition. And also impressing upon us the value of the technology coupling with the science. Dr. George Church is our next presenter. Professor of genetickic at Harvard med. He has spineered innovations in reading and writing DNA, he directs personal genomes.org with a goal of enabling open access integration of full genome sequences, environmental and trait data goal of working toward 100,000 individuals. Very interesting application. Again, this session being on applications, very eager. And Dr. George to hear what you have to say. Thanks for being here.
>> So thank you for the time here. As soon as my slides come up, I'm going to talk almost entirely about application. And it's going to different a little bit from previous talks in that I'm not going to talk about introductory definitions and in particular about what we can't do or having done but what we are doing. So this is my thank you slide. And my conflict of interest slide.
[LAUGHTER]
Next slide, please. Can I control this? There we go. Perfect. Okay. Sorry. And so as a graduate student, I work with Greg sutCliff, which we did a ridiculously high cost even though we were students, this has been used in many recombinant DNA efforts that some of them are listed here that we're really single gene efforts. What's wrong with this picture? This fellow is not using safety goggles. He's not properly grounded for electropouration. But the main thing is we've gone well beyond main genome engineering I had in the last slide. We have gone beyond minimal slides to these fast robust useful cells. We're focusing on lower costs. We have talked a lot about scaling up, but not lowering costs. I will focus on that. We look forward to whole genomes, but most of what I'll talk about is doing a little bit less than a whole genome but on a genome scale. And the question is, why do we do things on a genome scale? And then there's safety and security for the reason for doing things on a genome scale. And evolution is a unique capability that we have that most other fields in engineering do not have. And my major takeoff for all this is that we are going much faster than it appears. And we should not be reassured that biology is not capable of engineering and there's no difference between what we're doing and what I did as a graduate student. Why, genome-wide? We need to know why. Genome engineering is a commonly used term and is also a couple of genes done in the chromosome rather than on a plasma. Big deal.
Metabolic you might have a pathway or small network. 30 genes or less. But genetic code offers us multi-virus resistance and safety measures and some use of new amino acid and this is genome wide and one of the few articulated goals that is genome wide. The safety component is incredibly important. This is not meant to just be an analogy or images. But we have interoperable parts. These are all from cars but the same thing applied in biological design. Cost effectiveness, standards and isolation. We need to -- is it sufficient to have a set of rules and guidelines if there isn't testing, if there isn't surveillance? You can do licensing like driver's license but you have to do surveillance to make sure people are obeying the laws. And then again evolution is something that's new. There have been recommendations in 2006 and the next slide 2007 which I think don't go far enough. We talk about preferred practices. We pragmatically talk about federal grantees and contractors. There's a lot more out there than federal grantees and contractors. The Sloan 2007 went a little further than this. But we need to have surveillance and enforcement. And so back to my earlier recommendations on really licensing the entire ecosystem in synthetic biology, it's important. We need to have surveillance and testing of systems that are proposed to go in. And this is not restricted to bacteria. We have a very active human synthetic biology community and human do-it-yourself community. Some of my undergraduates have gone and sequenced part of their genomes on their own without F.D.A. approval and without really using any special equipment. And this is a whole another subject we're not going to talk about. But do it yourself or do it ourself biology and bio weather map and so on. We have studied vaccinations. Genome engineering, some success stories, we already mentioned one but also propane from DuPont to a $400 million project 90% successful. It only involved eight foreign genes plus 13 -- I'm sorry 13 down and six up regulations in the e.coli genome. 27 changes was a lot of work back then. I'm going to talk about hundreds of changes that we have incorporated. These are two other companies that I helped start that are not in the future but are already making thousands of liters of production scale fuels, either from biomass or from carbon dioxide and light. These are making alcanes, diesel and gasoline. Part of this is the success of comparative genomics. You can look through the bacteria for those that make trace amounts, Greg sort of alluded to trace amounts of the alcanes by taking fatty acids and reducing and decarbonnallating them. You can look at the genomes that produced and those that didn't produce these trace amounts and then you can identify the genes and overproduce them. Rob Carlson alluded to this exponential curve. This is actually quite different than his curves, although basically the same. What's different is that around 2004 or 2005, there was an increase in the rate of this exponential curve from 1.5 to 10 fold. And more importantly, this is a gap between the -- thank you. Between our recent huge increase in second generation or next generation sequencing and synthesis. And we're still stuck in the first generation for gene synthesis in the companies and genome synthesis that we're using first generation sequencing and synthesis for the most part.
There are 21 next generation sequencing technologies and 21 companies that go with it. And I am an advisor for about 16 of them. And similarly, there's a next generation synthesis off of chips that we've been doing since around 2004. This has lagged a little bit behind from making agains and genomes but it's certainly terrific for making short constructs.
Working in the cells, it's one thing to make DNA but getting the work in the cells, there are many tools. These are protein based specificity tools. And more general tools which are DNA based, they don't require specific proteins to put it in precise locations in the genome to make precise changes. But some of these involve single stranded DNA number 3 and number 4 in particular. And we have automated this in order to bring down the cost and extend our capabilities industrially. One of these is called -- or the general term is multiplex genome engineering or MAGE. And this has one particular implementation shown on this slide but there are many others. You can see it's a catch-all phrase. This one uses single strand nucleotides that use computer aided design to optimize secondary structures, optimize the position and length. You have to have a mismatch repair turned off for some of these. And there's a special proteins. But the key point is in a few years, we move from an efficiency around 10 to minus 4, 1 in 10,000 to 25% to 100%. And now we can get up to 8 mutations per two-hour cycle and we can just continue the cycle, 8 changes precisely in the genome wherever you want. And I'm sorry. You can make up to 1 billion different changes in a population. I'll show you an example where we did 100,000. This is Harris' prototype. A computer aided design of the upgrade. This is the sphul upgrade. This is applying it where we made 100,000 genomes, not one by one, but in a mixture. And it shows the awesome power of accelerated evolution in the laboratory, where we could make these 100,000 genomes focusing all of the changes in the known path ways, including putting in some genes from other organisms. And in three days, we can get the highest yields we have ever seen for this hydrocarbon lycopene which makes tomatoes red involved on the order of 24 genes. Another project that we have done which is less commonna torial and allows new amino acids and has safety features, here we changed all of the codons into TAA genome wide in order to free up that codon and allow us to delete the cellular factor that recognizes it. This can be generalized. There are 64 codons of these triplets and we have targeted nine of them. This allows us to do three things. New amino acids, safety features and multi-virus resistant which itself is a safety feature. We have these nine. We have done one of these nine codons that we're targeting out of 64. We have synthesized all the DNA to do the remaining eight, at least proof of concept on the essential genes. And another topic that is far beyond what we can talk about today probably is the project where we're making ribosomes and Greg aleaded to an in-vitro system which has interesting commercial applications. The key thing here is just changing these nine codons would require changing just 2.7% of the genome, not the whole genome. But if we're making these optimal 90, we have compiled the genome two and a half fold over and we essentially have remade the genome, even though we've only changed 2.7% of it. And that lies in the future, and it remains to be seen which is more efficient. Doing it all synthesis all at once where we'll probably have multiple failures or doing it one at a time. And just as a quick last slide or two is this issue of safety in terms of isolation. You can have physical isolation or you can have biological isolation. The changes of the genetic code, the genes can neither go out or come in that are functioning. The critics of the genetically manufactured organisms have wanted it.
A third way that it's isolated is physical and genetic and it's this metabolic dating back to the early days of recombinant DNA there was this acid that was used by deleting the biosynthetic pathway that you made the bacterium dependent upon that. It's not common in the environment, but it does occur. And that's one of the down sides. Some of these other SACB or tox-antitox pairs are used but as counter selections. But they are ways of having the cell self-destruct but they have the problem that they can be lost just before you need them. So they are not ideal. So we think going forward using the new genetic code to allow us to design multiple essential genes to have multiple dependencies that have been used in Peter Schultz's group. So in conclusion, just to remind you, you know, where we think we need genome engineering and synthetic biology, it's in making biology safer than it already is and this involves really using some of the lessons of other engineering disciplines, interoperable parts, hierarchyial designs, cost effectiveness, standards, isolation, testing, redundant systems, surveillance very important, not just surveillance of government grantees. Licensing at every part of the ecosystem. And focusing on this ability to evolve both in the lab and outside the lab. Thank you.
>> George, thank you for that. Your message is loud and clear in the face of advancement and technology advancement is astounding. And some near-term applications are very exciting. And also clarifies and I appreciate your last slide. And it was used before to help clarify for us what some engineering challenges are going forward. Our final speaker in this panel is Kristala Jones Prather. Dr. Prather is an Assistant Professor of chemical engineering at MIT and worked in industry as well as academia. Has been recognized for her work with numerous awards and investments. She is a research young investigator and received technology reviews TR35 young investigator award. She has also the NSF investing in her through an NSF career award. She's an investigator in the multiple institutional synthetic biology engineering research center funded by NSF. Welcome, we're pleased to have you here.
>> Thank you very much. Let me start by thanking the commission for an opportunity to come and speak to you today. The title of this panel is applications in synthetic biology. And what I'm going to do is try to give an overview in the field have been. And I hope what we can learn by that is both what we have done today and we can start to think about how that may project forward into what potential achievements or applications of synthetic biology might be in the future. Unlike George, I am going to start with a definition. You have heard a lot of them and you have heard -- I think what's clear is there is I will say lack of universal agreement on what synthetic biology is and how it should be defined. I'm going to give a practical definition, one we use within the research center. It's very simple and it says that synthetic biology is about making biology easier to engineer. You have heard some of these things before, particularly this morning. And in the first session about the relationship between biology and engineering and how they react with each other. For us in particular, it's about applying engineering principles to biological testimonies and it involves words like design, modeling and characterization. I was raised by Jay Keasling and there's a well-known cartoon that Greg Stephanopoulos used to show that there's a group of students in the class and the student raises their hand and says what's the difference between metabolic engineering and genetic engineering and there's a professor that says lots and lots of math. And then there's a picture of the professor and no students. There is this idea that we like to have models of systems that are numerical and mathematical. And it's an attempt so we can have this loop back and change your model and see what the new characteristics are. So I think that is a part of synthetic biology, which has traditionally been different from genetic engineering. But it's not wholly distinct from what you may know as systems biology. Again this effort to include math and the ability to predict and design and what we do.
And we'll highlight DNA synthesis as an enabling technology. You'll see from the first few slides that if we're talking about making biology easier to engineer and we want to get started with that now and based on I thought what Drew gave was a very good slide of the technology gap, if you will, between the ability to write DNA and to know what to write on DNA. Much of what's happening now under the umbrella of synthetic biology is using DNA synthesis at a very minimal level because we have to start with some existing biological substrate. In that vein, if we think of this goal of synthetic biology and what is our biological workspace, we heard about microbes being the substrate of choice because of their relative simplicity and I'll use relative quite intentionally because we're still talking about very complex organisms although they are less complex than the million cells which you see there and also plants. And if you think about now from an applications perspective, the biological substrates we work with, the applications I think become pretty clear in terms of extrapolating. We can think of therapeutics including pharmaceuticals as well as biologics or biopharmaceuticals and protein therapeutic fields, energies and I'll give a brief slide on that. Chemicals which may be part of the pharmaceuticals but leading toward thinking of new ways for materials to have renewable materials, things to get rid of other polypropylene bottles which will fill landfills if we can't figure out good ways to recycle. And agriculture when we think about the biological works of plants and the potential to expand with genetically engineered organisms for agriculture. Wrong button. Too many buttons. With this paper here that we have heard about already, which is the work from the Keasling lab from the University of California at Berkeley, producing the antimalarial drug which can be used for an antimalarial, this was funned as the numbers have come up. I am sure we can all recite them. $42.5 million from the gates foundation as something of a public-private partnership between UC Berkeley and Amyris which was a company founded by folks from the Keasling lab to develop this technology. One of the unique aspects, intellectual property came up previously. There were lots of issues because the University of California had to agree to make the licenses available essentially free and the commitment by all parties involved is that they would develop this as a remedy for at-cost production. This was to be a non-profit generating venture as far as the company is concerned. Amyris, if you have been keeping up with the literature, has sort of transitioned this process. It's now in the hands of industrial manufacturing and they have switched their focus almost exclusively to fuels. So it's an example of how the basic technology of these achievements and what we're able to do with engineering of biology with synthetic biology, with metabolic engineering, whatever particular phase you want to use, builds a repository of intellectual information and intellectual properties that can be then converted into other downstream applications and in this case from therapeutics to fuel. We have talked a lot about microbes. That work was done in microbes. There are efforts and achievements in synthetic biology going into increasingly more complex systems. This is a paper from Martin fussnegger's group about developing effectively a circuit to control gene expression for implants. So the idea was they were able to take pieces from microbial cells to respond to a particular molecule they put into a skin lotion. They could have subcutaneous implants. If you applied this lotion, you would get gene expression. This notion of a circuit to control expression of a gene from the introduction of a small molecule, this is now an example where we can think about how that actually has potential applications in medicine in terms of being able to activate gene expression perhaps with novel forms of gene therapy that would be a subcutaneous implant so you're not talking about trying to modify the genome with more I would say complex perspectives of gene therapy where you're looking at, for example, removing some cells and reengineering them and putting them back into the cell. This would be a separate implant that would be distinct from the native or the human chromosome. Moving on to the field, I have already mentioned Amyris work. This is work from the University of California in Los Angeles that was published in "Nature" a couple of years ago for making higher order branched alcohols as biofuels. This is technology licensed by GeeBo. Dr. Bassler mentioned and if I can paraphrase from going to scale and optimization and getting something industrially viable, this is the case where this work was licensed by a company and they are actively working to commercialize this process. Similar to the work done that we have heard about before, these are pathways to a certain extent are all natural. The molecules being produced were ones being identified as minor products in wine fermentation so the enzymes or the genes needed in order to convert what ends up being intermediate amino acid synthesis were optimized in the most promising was 22 grams per liter of iso buttenol being produced. This is a screen shot from a website. I wanted to highlight the fact that they really do talk about themselves as being a synthetic biology company, being able to take advantage as one has already referred to of all the extensive information that's come to us from genome sequencing projects and increasingly the tools and technologies that we're developing and being able to take advantage of that had have to do with synthesis and construction of biology. They are focusing on fuels, but also on biochemicals. This is an example I mentioned before in terms of energy but not being fuels. This is a paper from April of this year from a lab at MIT where again because of the multiple definitions of synthetic biology we may or may not think of this as synthetic biology. But it just describes briefly what was done here. The Belcher lab at MIT used M-13FHAGE as biotemplating devices. They were able to use them and the PHAGE interact with inorganic often metals and able to form these higher order structures. This is a case of biological expression and biology as a template for making these nanostructures. We could certainly think about how to expand that towards now having the power of synthetic biology and constructive biology to be able to redesign these PHAGEs so the structures are complex. And the particular application was to be able to put together a structure that would allow you to have effectively photosynthesis. And they had the idea you could use this for energy storage and capture the hydrogen from the splitting of water and that hydrogen can be stored and used at a later time. And solar energy you have available when the sun is out and don't have it available when the sun is not out. This is work from my own lab in collaboration. In the chemical space what we were looking at is being able to make a pathway for a compound acid where we don't actually have a natural metabolic pathway for this compound. This is different from the work I presented previously on the branch of alcohols where we weren't starting from a pathway and trying to reconstruct. We started here's a compound we want to make, how do we think about doing that? The particular innovation in this case was to be able to use these novel synthetic scaffolds. And Dr. Bassler mentioned the wonderful spatial organization that happens with a naturally occurring system. This was a synthetic device designed to introduce this spatial organization into a microbial cell. And the result was to have increased productivity for the compounds we were interested in. And I want to refer to biological computing or a lot of the analogies to programmability. The first of which was a program about 10 years that described the repress later. And also from Jim's group, the first was from Princeton and Mike Elowitz and the oscillator called the retabbilator, which is often a fluorescent protein this is oscillation in the metabolite. Now going from again microbial systems and mammalian cells and this was referred to by Dr. Bassler and is now looking at these oscillators and genetic clocks taking advantage of intercellular communication. And this is often discussed and deresided as toy applications and you're just making cells blink. What is that good for? From my own perspective from making these that make high quantities of synthetic chemicals, we're interested in these because we know that timing of gene expression is important for some systems that we're looking at. So we can look at oscillators that have been designed even with clean production proteins and think about how do we extend those into practical applications of systems where we're using them either in therapeutic purposes in order to have time expression of genes, for example, in development, talking about stem cell biology or even in like a large bioreactor talking about chemicals. The last sort of screen set I have is the paper that again was sort of the impetus for this particular discussion from the Venter group which you have already heard about. And I just -- my comment I wanted to make sure is all the things I have talked about so far, you may be thinking what does that have to do with synthetic genomics and the ability to completely synthesize the bacterial genome, what I would say is this is about trying to bridge this
technological divide. What we currently have is the capacity to do very extensive reengineering of genomes from existing cells, taking out lots of genes. Putting in lots of genes. Beyond that, where the challenges often arise or how do you precisely control them, temporally, spatially, these other issues about natural biology, they are complex and very confusing for us. What you have here is now this very clear synthetic capability. And where I see this bridging is that as we get better and better at understanding how to do the kinds of engineering we're doing, then it really is about the differences in scale that Dr. Bassler referred to this morning, that we can think about now going from making these manipulations at the level of an existing genome towards designing them in de novo and starting from scratch with a genome that works the way we want it to work. The final comments is there are, of course, lots of challenges. Biology is complex as we have heard over and over again. I'll add it's often context dependent. We do have the stream of having interchangeable interoperable parts. I'll say from personal experience, you move them from one cell to the other, they don't work the same way. And that's exciting. It's a challenge. It's something that we have to become better at understanding. The synthesis capabilities as you've already heard far exceed the technological capabilities and that's a gap that does in some way point at what our future ambitions are but indicate what our current limitations are. The potential benefits I think are enormous. I indicated a few of these, but, you know, we can think about this in any way where we think about biology being important. At the same time, the risks are real. Because there is this information gap between what we really understand about biology and what our capabilities are, it's impossible for us to really predict what's going to happen in every single experiment. And so I do think it's very worthwhile to think about being as careful as possible as we do this to minimize those risks. And two seconds over, I'll stop.
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>> Very impressive. Thank you very much for that list. And also ending with a challenge that we have ahead. Keeping with the format we used before, I have asked the commissioners to get their thoughts together. But I'll return the favor, Amy, if you would like to offer the first question.
>> Thank you very much. And thank you all. Let me begin with a question to Craig, if I may. The potential power of synthetic biology creates hopes and it creates fears. And we're all too well aware now of the fears. But I want to begin with the hopes as well. So you mentioned the one-day production of a vaccine for flu, for example. So here's my question to you. What is the single hope that we should most believe in from synthetic biology moving forward? And it would only be incumbent on me to ask you the same question with regard to fear. What is the single fear that we should take most seriously?
>> Well, they both give me wide latitude, so I appreciate that. I think --
>> Don't make it too wide.
>> On the hope side, obviously, what our own teams and others are trying to do as well, we need new tools to make new medicines a lot faster. Particularly with vaccines. It took quite a while with H1N1 to get a proper response, in part because the rate of building and deciding on seed stocks and in part because we're using 100-year-old technology with chicken eggs to produce vaccines. Both need to change and quickly. But with rapid sequencing and all these changes in reading the genetic code, and now the ability to quickly write the genetic code, it's now hours instead of weeks and months to make new seed stocks. The potential applications because we can design cells with hundreds to thousands of energetic variation, diseases like HIV that they were chatting about with that change their genetic code very quickly. The rhino virus, we don't have a vaccine against the common cold because the virus evolves rapidly. Designing things with the same rate of evolution or covering the spectrum of energetic variations gives us whole new ways to approach vaccines that never existed before. On the environmental side, I think it's clear we need to do something different in the environment as we go from 6.5, to 9, to 10 billion people. We can't keep doing what we're doing. So attempts, all these different attempts, they all need to be successful in creating new sources of fuel and energy and food or humanity will be irreversibly damaged and altered. So we are a society dependent on science now for our future. Biology is a key part of that future science. Synthetic biology, synthetic genomes are key I hope components of altering that future. On the fear side, obviously, the worst scenario is what happened in computing because we're talking about software. People make computer viruses that cause a lot of economic damage. Well, we don't want the same mentality going into making new animal or plant viruses. Either inadvertently or purposely. And some of that can be readily prevented by some pretty straightforward regulations. But obviously, nobody who develops new technology wants to see that ever produce harm to others. We just would like to see just the benefits. I think the molecular biology community has a pretty good track record for the last several decades because of the guidelines and rules that we have all been working under.
>> So I also want to direct this question to Dr. Venter. I heard both your views and in the literature people have talked about the publication of science and the proof of concept. And I wanted to understand exactly what it is that it is proving. In part, as I understand it, the cell wall of the bacteria was used in the first generation and it's a natural organism that has been synthesized. I'd like to understand what it is that it proves and how significant that proof of concept is. And second, building on that, looking forward, I understand that you may be working on algae and other multi-cellular organisms where the genetic information is in the nucleus of the cell rather than a single strand.
How far away from that are we? Is that the proof of concept that will propel this field forward?
>> What's been possible in molecular biology is what several people have described this morning. Changing one or a few genes in the cell by inserting the genes in plasmics. Although some evolve by taking up chromosomes, for example, color has two chromosomes from two very clearly different origins so they probably happen through these kind of processes. But never before have we molecular biologists been able to take an entire bacterial chromosome, an entire chromosome of anything other than a small virus and transplant into a cell of one type and convert that cell into another. Then you add to that, starting with the digital code in the computer making the entire compromise from scratch means now we have the means to start with that digital code, make gramattic changes. While we'd the basis of an existing organism, we made changes to it and inserted the names of 46 authors, several quotations. It's the first genome with its first built-in website and web address. These may seem like trivial changes but identify it as a synthetically made chromosome something we think is critical for this field. And activated that and completely transformed that one cell into a new cell. It was not trivial. One base pair being wrong set us back three months. One error out of a million base pairs did not enable this to happen. So it's now, because it's a proof of concept, we do know how to do it. And now we can make much more extensive modifications. So we're building a robot to do common synthesis instead of making one chromosome over 10 years, our goal is to make 1 million or so a day by randomly sorting genes or selecting very specific ones, selecting living cells that you can't get. It's not a species that existed. It's very closely similar to a pre-existing cell, but it grows substantially faster because of the 14 genes we eliminated.
>> And on the multi-cellular front.
>> There are a lot of eukaryotes, moving nuclei around has been done 50 years or more, changing the DNA in the nuclei and replacing the DNA we don't think will be a huge challenge. It's probably usier to replace the chromosomes and eukaryote yeast by replacing them one at a time with synthetic DNA.
>> Thank you.
>> Dr. Venter, do you think it's fair to say that, you know, in the very elegant transformation experiment, that really that's how I read your paper first on that day, I saw it as, you know, probably the world's most elegant bacterial transportation formation that had been done to date.
>> Thank you.
>> I think maybe trying to clarify what Nita was driving at, you need today collaborate with existing life in order to make that transformation experiment work. And whiem it's true that after after several replications that all components, not just the proteins of that cell, were obviously derived from what you had produced in sillico and printed out, it did require a collaboration with existing life that had been derived by natural selection.
>> Absolutely. So we're starting, as I said, with the 3.5 billion years of evolution. We used that starting system to read the new genetic code and start making all the new proteins. As I said earlier, I think it's an interesting scientific question how few of those components we can get away with. As I said, perhaps just a ribosome, some polymerase, a few lipids. So, you know, when people evoke that you start with existing life, it takes us back to vitalism, that people try and amazingly, the "New York Times" has tried to reovoc vitalism. Most scientists view it disappeared 8 years ago as a concept and certainly with DNA being the material coding for everything, there's nothing vital in the cell other than the ability to read that new software. So we are clearly software-driven machines. That software is DNA.
>> First, thank you all very much for your comments. I would like to ask Dr. Venter, this is an important scientific step, but as you described what you have been working on for many years, you also described a process of thinking about the ethical issues right from the very beginning. So I am wondering if you could say from your perspective, what has changed now ethically, if anything. And where you think building on Dr. Atkinson's question earlier, where you think -- I think you mentioned we need an intelligent ethical-legal framework. Where are we lacking in that regard? What do you think we can do to help in that regard? I'd love to hear others' opinions on that as well.
>> I think it's a very critical question. It's not clear that anything has changed so dramatically as what some people describe as minor changes in biology with minor but significant changes in the ethical and legal framework, primarily because the way we control who has, for example, A-list agents and has been controlling who has access to these agents. Now, if all you need is the genetic code in the computer, it totally changes who has access and how you get access to them. If students can order anything from a DNA synthesis company and there's no tracking of what they order, some could try and make ebola virus which is only 8 genes or at least the DNA. The DNA is not ineffective but I'm sure if Homeland Security started detecting an ebola virus DNA, they'd probably get upset. Those would be the kind of hacking things that we don't want to occur. I think those can be pretty much eliminated by requiring companies to screen against A-list agents and requiring bona fide institutions to be doing this work versus being done in somebody's garage. I think creating new life forms -- I think what we did is as much a philosophical step as a scientific-technical one. Because it now opens the window for literally merging the digital world with the biology world. And because anything that's totally open-ended, we think there's some guidelines that are needed. I think it's sensible to start in that framework, so that we don't get the negative consequences are unintended ones from lack of paying proper attention.
>> George, you have written on this as well. Would you weigh in, please?
>> Yeah, I'm not sure whether this is an ethical or policy issue. But many of the previous discussions, the conclusions have been we should have more discussion. And I think that we are actually in a place that we can do more than that, which is to focus on licensing and surveillance. And I don't know whether that's a new -- whether that's ethics at all, much less a new one. What's happened since 1999 is this exponential curve has gotten steeper. And I think that's something you can't ignore. So I would say that it's time to go beyond having more discussions.
>> Following actually on that, it seems to me we have heard a lot both from the previous panel and from the three of you about the widespread availability of various codes, the quote-unquote ability to do it in your garage, unquote. But that seems to me to refer to the obtaining of the sequences and perhaps the synthesizing of those and that generates worries for people. But the question I have is, how big a step is it -- and you have alluded to this, I think, Dr. Venter -- from having the sequence to actually getting it to work in a biological system? And is that gap big enough that we shouldn't be as fearful as we are of the possibility of this being misused because we could in fact have regulation or safeguards at that step that would be very helpful? True or false.
>> That wasn't a yes-no question. I'm sorry.
>> You have a 50% chance of getting it right.
[LAUGHTER]
>> Then I'll say false. I think with each new cellular system and the micro plasmas don't have a membrane which made it simple to get the DNA across. What we are trying to do with the algae right now is maybe using a plasma membrane to transform things. We're at the earliest stages. We need to see how extendible these tools are. Getting DNA past cell walls may be very tough, but there are other ways to get around things. The two areas go in parallel. One is the design and the synthesis and the two booting it up. The biggest worry was we were going to have this really nice macro molecule, the largest one of a defined structure ever made and we couldn't activate it in the cell. We were there for a long time because of one single error in the genetic code. So I think it's going to have to be optimized for each individual biological system. It's totally different getting DNA into plants than it is to bacteria and totally different with cell walls, without cell walls. What I think this is going to be a rapid expanding area of research and probably difficult to regulate. I think the guidelines that get set up for approving projects at the institutional level with broader guidelines at the funding level and even though our work was not federally funded because my institution is a major federal grant recipient, we have to follow the federal rules regardless of whether it's funding that particular research. So I think the way molecular biology has been practiced, particularly in this country, has been I think a wonderful example of how to proceed, but expanding the repertoire and expanding some of the ways we monitor things.
>> Go ahead.
>> Yeah. I may be misinterpreting the question. So the information is free. You go to the database and get as much sequence as you want. It is cheaper, but still not trivial to actually pay for synthesis. So my lab does not yet, as a matter of pract tis, synthesis everything. We still do a tremendous amount of PCR. I just had a meeting with a student a couple of days ago and said, okay, you can get these things synthesized and it's going to cost about 3 thowshd but you can't get the 12 other variants of it synthesized that you want. Now, we're talking about $36,000. So as far as access, some of it is thinking to the future in terms of if we go -- certainly we're not at $10 a base as George showed but we're under $1 per base but not at a dime per base, at the level of small amounts of orders. So you can do negotiations with some companies to get things on the order of 10 cents to 25 cents a base if you want a lot of sequence. So because of that, I think again I may be misunderstanding your question. But I think there are different answers in terms of whether or not you're talking about institutional access versus noninstitutional access, skilled labor versus unskilled labor. In terms of access because of the cost, I still think that a lot of the more fear was the word used earlier but the things that may evoke fear and apprehension are still beyond the cost of most noninstitutional players. And then because we're really talking about difficult biology and one base pair mistake setting you back three months, there's still a big difference between what you can do as a skilled practitioner versus an unskilled practitioner.
>> Just to follow up, I appreciate hearing that it's a little more difficult than just doing it quote-unquote in your garage to get the sequencing done. But I was talking about the next step and whether people can do that in their garages quote-unquote, of getting that to replicate inside a cell and how difficult that is and what material is needed there and whether that's an important place in which regulatory safeguards could be placed to make sure this doesn't get into the wrong hands.
>> So, at the simplest level, and if you read some of the blogs and the popular press, everybody wants to make things glow. You want fish that glow. And it's like let's put protein in anything you think about. It is relatively inexpensive and on a skill level relatively easy to order a gene that would affectively be a plasma that encodes for green fluorescent with motor on one end and terminator on one end and transform a bacterium in your garage and say, hey, it glows. It's very difficult to make your dog glow. So again, we're still talking about a level of complexity there. And the one gene being able to transfer one gene and getting that to work in a garage with a junior high school student, pretty close to trivial. The types of things that the Venter lab did, not going to happen in the garage with 14-year-olds.
[LAUGHTER]
>> Thank you very much for your presentations. All three of you really talked about the need for some level of regulation. And I wonder if you could comment on whether all of these different things that fall under the definition of synthetic biology are already covered under existing regulations because certainly we have regulations of how to handle anthrax or ebola or other this process of things. Do you feel we need to have different and new types of regulations to deal with the issues of synthetic biology?
>> We certainly have recombinant DNA regulations. Many of these depend on the person practicing and having federal grants or in some other way being a responsible citizen. I think what we don't really have is surveillance that the regulations are being obeyed by all citizens, not just the standard members of society. And I think we also don't really have many regulations about safety testing as we make things that either are intended or could accidentally get into the environment. I think as safety testing, we take for granted in many other engineering disciplines, there's relatively little of that in biology. It probably doesn't require major overhauls but I think there are some gaps that we need to pay attention to.
>> There are really no limitations on what you can order from a nucleotide synthesis company. At the present time, they are not required to screen against any list of agents. Some are voluntarily doing it now. And it's not just a U.S. problem. DNA synthesis is a global effort. If you can't get what you want here, you can order it in Germany or you can order it in India or get things made in China. You can buy DNA synthesizers off of eBay. So maybe there are, as people said earlier, four companies that are probably 90% of the synthesis in the U.S., even though they are not all in the U.S., requiring them to screen against A-list agents, requiring them to have bona fide credentials of the ordering institution, I think are things that could go towards preventing the frivolous use. There is a lot of home-brewed biology being done in kitchens. It's a new trend. Drew Endy has been in the past -- I was pleased to see he stopped doing it and encouraging biohacking. You know, we want some reasonable restraints on that, without destroying this wonderful creativity that these kids are doing to come up with some new circuitry that could totally change what we work on. But I don't think it's covered by any of the existing regulations.
>> Just as a follow-up, you know, in the kinds of experiments that you published, it is possible to be able to take what nobody may be able to recognize and may come from an organism and you could order a bunch from one company and a bunch from another company and be able to put together.
>> At the level of 100 meres or anything over probably an 18 mer or something, you could get a pretty good trend of what somebody was trying to do. The signatures are pretty clear-cut.
>> Also, these companies are beginning to coordinate voluntarily. This is something that would be nice to be backed up with regulation. But they are voluntarily coordinating their efforts. So if someone split their order over four companies, that in and of itself would be an alarming event, which combined with the sequences that could be recognized, I think you could put the story together. But it will be ongoing efforts to get around that.
>> How quickly will you put the story together realistically?
>> Well, if it's entirely based on computational algorithms pretested -- and I emphasize the importance of testing. You could put it together in hours. The computers, especially if you have got government agencies that are willing to act in hours.
>> I think you have gotten to the heart of a lot of my concerns. As you probably all know, I'm there was a strain of anthrax that seems to be becoming a low hurdle to overcome in the ongoing biological processes.
I have heard a couple of different things. One is that this is still difficult to do, very difficult to do. But the second part is that it's getting easier. And so I think that raj brought up an excellent point. There are regulations in place now for I think what we would consider traditional biology being able to reproduce organisms that are on the biological toxins list. However, we're talking more about the biobricks now which, quite frankly, are not part of the regulation. So what concerns -- and I think you have expressed them here. So your concerns about this evolving technology and getting around the B-sat, the security measures that we would need to take to make sure that these would not happen, and what is the balance? You have probably been privy to the discussions of the latest B-sat. In balancing scientific discovery versus security for the American people.
>> I don't actually think that this is a trade-off between security and scientific discovery. I think if this is properly implemented, where most of the effort is in developing computer software and getting compliance at the company level and getting surveillance at the government level, the researchers in a certain sense shouldn't even see it. It should be transparent to them and they can get on with their work. On the other hand, if you require them to sign a piece of paper every few minutes and every time they type something, you could interfere. I think that's unlikely that's where we would be going with this. I think some serious computational efforts are in order.
>> I am interested in the money behind it all. It's a very expensive proposition now to come up with a new cell, as you did over $40 million or new products. And mostly funded now in often small biotech companies and with venture capital kinds of money. Are there recommendations for being able to encourage the entrepreneurship, while not having so tight control on it that you can't get a payback to the amount of money you spend putting into a project and can't get ta payback fairly quickly? Versus being able to also encourage entrepreneurs to work on projects. I mean there's going to be huge profits in this, if it would work out the way it is looking like it might work out in biofuels or energy and so on. I just wondered if you had recommendations or thoughts on what the commission should recommend on those issues.
>> Just my opinion is the current system is actually quite healthy. In contrast to the one that Rob Carlson described, most of my experience with dozens of companies is they can get the job done without spending a lot of money on lawyers. Very often you don't really even need the patents in the end. It's the know-how that's incredibly important. I have very few examples of a patent getting in the way of academic research. And generally, not even in the getting in the way of start-ups as well. This is such a vibrant field that people are inventing so quickly, that they invent around or don't even concern themselves. I think it's actually quite healthy and going from small to large is happening quite quickly, too. Craig mentioned Exxon and the case of LS9 and they have Procter & Gamble and Chevron. This is in theory a authority number of years. I think in my opinion it's healthy.
>> In fact, if I can add briefly to it, it's healthy and critical. I think if all these bets are right that everybody is placing to get proper ecological benefit and change the use and dependency on taking carbon out of the ground and burning it and putting it into the atmosphere, we need things to work economically. And I think there's a healthy investment climate in the U.S. despite the stock market. Better step in where the government hasn't.
Most of the advancements in biotechnology have come with companies like Genentech. In our case, we would have been stuck back in 2003 with a small synthetic virus if we did not have independent money from starting synthetic genomics to fund this work at the not-for-profit institute.
>> I would only add to that I think one -- so I agree with what's been said. I think one of the impediments to progress, if you will, that can arise if all of the achievements are done individually, is one of the very big goals of synthetic biology is to have standardization and interoperability. One of the ways the federal government can help with that is promote in some tangible way an effort for the community to be able to organize on a regular basis around what should those standards be, so that you don't have innovation happening in isolation in a way that you have very great technologies evolving independently and to network those and interface those becomes very difficult. As we dream about synthetic biologies and you see the leggo kits all over the place as a good analogy, that works because you have standardization and you know you can get leggos from anywhere and they are going to work together. That's an effort I think has been more difficult to get real support for. Because it's not -- it's fundamental. It's foundational. And it's enabling, but it doesn't in and of itself get you biofuels and it doesn't in and of itself get you new vaccines. It facilitates all those things and sometimes there is a gap between the foundational more engineering-oriented standardization work and the applications-oriented things which can be very interesting and attractive to investors.
>> I want to begin by thanking all of you three for excellent presentations. I have got a couple of concerns or questions for you. One has to do with the fact this is all now accessible on the Internet and it's international. So if we're a commission set up to think about regulations here in the United States, I'm wondering what the context of our deliberations should be. If these activities are really taking place all over the world. So, you know, I mean what sort of international collaboration has to take place for U.S. regulations to have any real effective bite. That's the first question.
>> I think it's a critical question because science is international. These tools are international. The Internet is international. And I think first and foremost, the U.S. can set a good positive example. That didn't happen with stem cells in the recent past. And research expanded overseas at the expense of research in the U.S. I think we can do the opposite here if we do it intelligently. The same concerns that we have here have been expressed in the EU and basically every country I visited around the world. So I think if there's a positive example of how to deal with things, that would be a good start. But it has to be international ultimately to have any impact.
>> Thank you. A follow-up has to -- it's really a follow-up question about the role of industry. There has been some talk around the table about the movement from small to large, right. So listening to all the panelists, you get that picture that we're currently living through, the kind of biological Woodstock with people experimenting in their garages and so forth. But the movement will be, as it's been in the pharmaceutical industry and computer industry, from small to large. And so I'm wondering what the implications of that might be with regard to access to the goods produced by the industry. We have seen in the area of pharmaceuticals, a lot of public concern about the patent system and the rules and regulations relating to access, particularly with regard to access to life-saving drugs for diseases like HIV, where it's perceived by many people that the patent system is working against access to life-saving medications. So I'm wondering if we have anything to worry about that's analogous in this area, right. In other words, should we be worrying now about the synthetic bio analogs of Microsoft and Pfizer limiting access to knowledge and limiting access to the goods that are produced.
>> My answer is quite simple, no. I don't think there's any worry at all. In fact, the worry is in the opposite direction. If we don't get the things that really work at a commercial level, this is an interesting academic field. Published in a paper like my team did in science is great for understanding the concepts, converting it into reality where you can buy fuel at the gas pump made from carbon dioxide instead of from oil out of the ground will only work if that's done in an economically competitive environment. LS9 and these companies, synthetic genomics will only survive if they have economically competitive products. Most people aren't going to buy things just because it's better for the environment unfortunately. So any new fuels, for example, have to be available and they have to be cheaper than existing fuels or at least cost competitive with it. So we need economic driving forces to pull this stuff much more rapidly than is currently happening. I don't see any limitation of access. We need access pretty rapidly to CO2-based fuels as an example.
>> Before I go to Nita, I don't want a thought that was dropped, Kristala, that you brought up and connected with something to George. There's the flip side of regulation. And that's stimulation. Will one of the effective ways to ensure safety to be the sort of a way to be able to skate ahead of the puck, know where the puck was going? Were you suggesting Dr. Prather -- maybe I should leave it open. The question that we had sort of posed in the prior panel to you.
What would be the very next thing to be funded? In view of being able to have a knowledge base and an ability both to advance the applications of the kinds you have all been talking about, but have a deeper knowledge to help ensure that we can recognize as Dr. Church mentioned some of the sinister potentially sinister applications of these things. What would you fund next? Would it be your standardization?
>> You're not allowed to answer your own lab.
>> Yeah. That's fair enough.
>> What's the second thing?
>> No. It's a difficult question. So I think about, for example, the biofab which Drew Endy is directing which has the ambitious goal of being a focal point where you can develop parts to use a term in synthetic biology, discreet pieces of DNA in code for some biological function. And you can characterize them and see how they behave and see things like composability, what happens this this thing and the other thing. And I think it's a very ambitious goal and I think it's great. And they have got like $2 million in two years. And so what happens when that's gone? So I think that there's a need for an effort that is more ambitious in scope and much bigger in scope to say, okay, let's bring -- it does two things. One is that it can serve as a forum, if you will, for bringing different players together and brainstorming and saying, okay, here's what I'm doing, here's what you're doing. Here's what this person is doing. How do we get those to interface in a way that we can actually set a standard moving forward so that as new technologies are developed, we know they are going to fit in very well? And then how do we set priorities for -- and safely, yes. And so I'll jump ahead to a thought. You know, I think one of the things that's been very nice about the sinberg experience is we have had discussions all along about discussions and the thrust called human practices that deals with biosafety and biosecurity and intellectual processes. But that is a very, very small number of people. And what's becoming an increasingly populated academic field.
If you look at the number of people associating themselves with synthetic biology, it's grown astronomically over the past few years. And so there are questions in terms of if you are focusing and if you take NSF's investment into synberg and take DOE into the joint bioenergy institute, you're talking about real money. We're not giving it back. It's not trivial but a very small number of people it's impacting. I'd like to see efforts to bring the community together in a way that we can think about what the next steps are going forward and that we can be more progressive and proactive as opposed to reactive in saying, well, you did it wrong, so here's my other way to do better. We're in that midst now where it's a bunch of you go back and forth between I did it this way, that way is all wrong. Here's why it's all wrong. And I'm not saying that won't eventually get us to where we're going. But if we want to be able to bridge this technological divide and say we have exciting technology and the potentials for the impact it can actually have on human existence are very real, we need to be able to move that forward more quickly.
>> Thank you. Craig, did you have a comment?
>> The two questions I get more often, most often when I give lectures on this topic is people are worried about bioterrorism and environmental release. And it depends where you are which one is first or second. And so George gave some wonderful examples of safety mechanisms that could be built in. It would be nice to have orders of magnitude more. We're trying to build in suicide genes to organisms. If we're going to have large algae plants made from genetically synthesized or modified organisms, they need to not be able to survive in the environment on their own. Suicide genes, chemical dependencies, using artificial amino acids so they couldn't possibly grow in different environments, expanding the repertoire of what is safe and secure I think would be the sing the most beneficial thing out of any government funding.
>> I'll just quickly -- and I think coupling this question to the previous small versus large. Small has safety advantages. It's only once we got to the large manufacturing of automobiles that we really started getting very high levels of safety. And furthermore, as the technology gets to a certain point, amateurs stop making it. So, you know, I made a computer when I was young. I wouldn't bother to make one today. The know-how starts to fade away at the grassroots level which is a mixed blessing. But from a safety standpoint I think it's incredibly important.
>> Certainly since we're taking so much of a lead from engineering, it made me very proud of my heritage actually today. But one thing that has been demonstrated in so many physical systems is that it is far more effective to design in safety than it is to try to regulate in safety. That was the basis for those questions and very responsive. Thank you. Nita, I think you're next. Anita is next?
>> Thank you. I really have appreciated all these remarks. I wanted to ask Dr. Prather a very specific question about something that you said toward the end of your slides. You had a slide in which you made the intriguing point that our synthesis capabilities exceed our design capabilities. We know how, but not what. Could you elaborate? The reason why I want you to elaborate is because I'm wondering if we know how but not what points to some limitations on the applications that may be forthcoming from this science.
>> It absolutely does. Simply put, this is all back to comments that Bonnie Bassler made earlier that these are really complex systems that we're talking about. I don't yet know anyone in the field for whom their design works the first time they implement it. And the question is always -- I describe when I am sort of giving my pitch to first-year graduate students, they say what we do is to pick molecules we want to build and then we have problems. And your thesis research is all about how do you solve those problems and what do you learn from solving those problems. And we often learn things we didn't expect to learn. Sometimes we run into problems and they go, yeah, we figured that was coming eventually. There is two different aspects of it. One is that biology even for very simple organisms is still very complex. So being able to predictably know what's going to happen if you make a single perti vaition is difficult to do. That's one aspect of it. The other part is the type of manipulations that we're talking about doing are in and of themselves somewhat different from what we see in nature. We are mimicking nature but trying to take natural components and stream them together in ways that haven't been done before. There are no some cases a lack of fundamental knowledge of how that's going to behave. So there's a need for experimentation, to actually have the observation that says, okay, this is what I observed when I did this particular configuration. Let me make four or five different variants and see those observations and then put it on a graph and see if it's just a random set of points orphic then draw some conclusion that if I have these specific changes, here's the effect I'm going to get from that. What the Venter lab group I think has shown that the capability, the capacity to go from sequence on a computer into something that is physical DNA is there. But if you ask anybody, okay, you're free to write 500,000 base pairs, DNA, what would you do? Most of us are going to copy something that already exists because we just don't know how to make it all work together. And even the stuff that's working together, we don't know why it's working the way it's working.
>> How can we get better at knowing what -- given our genius at knowing how, how do we get to the knowing what.
>> There are two parts that also came through this morning. Some of it is more information about biology. As an engineer, I have no desire to interfere with the biologist doing what he or she does on a day-to-day basis to uncover fundamental knowledge of biology. I applaud it and we steal as much of it as we can with proper credit that we took it from somebody.