At the break, I was asked by several people
about my comments about the aging debate.
And this will be my only comment on it.
And that is, I understand
that optimists greatly outlive pessimists.
(Laughter)
What I'm going to tell you about in my 18 minutes is
how we're about to switch from reading the genetic code
to the first stages of beginning
to write the code ourselves.
It's only 10 years ago this month
when we published the first sequence
of a free-living organism,
that of haemophilus influenzae.
That took a genome project
from 13 years down to four months.
We can now do that same genome project
in the order of
two to eight hours.
So in the last decade, a large number of genomes have been added:
most human pathogens,
a couple of plants,
several insects and several mammals,
including the human genome.
Genomics at this stage of the thinking
from a little over 10 years ago
was, by the end of this year, we might have
between three and five genomes sequenced;
it's on the order of several hundred.
We just got a grant from the Gordon and Betty Moore Foundation
to sequence 130 genomes this year,
as a side project from environmental organisms.
So the rate of reading the genetic code has changed.
But as we look, what's out there,
we've barely scratched the surface
on what is available on this planet.
Most people don't realize it, because they're invisible,
but microbes make up about a half of the Earth's biomass,
whereas all animals only make up about
one one-thousandth of all the biomass.
And maybe it's something that people in Oxford don't do very often,
but if you ever make it to the sea,
and you swallow a mouthful of seawater,
keep in mind that each milliliter
has about a million bacteria
and on the order of 10 million viruses.
Less than 5,000 microbial species
have been characterized as of two years ago,
and so we decided to do something about it.
And we started the Sorcerer II Expedition,
where we were, as with great oceanographic expeditions,
trying to sample the ocean every 200 miles.
We started in Bermuda for our test project,
then moved up to Halifax,
working down the U.S. East Coast,
the Caribbean Sea, the Panama Canal,
through to the Galapagos, then across the Pacific,
and we're in the process now of working our way
across the Indian Ocean.
It's very tough duty; we're doing this on a sailing vessel,
in part to help excite young people
about going into science.
The experiments are incredibly simple.
We just take seawater and we filter it,
and we collect different size organisms on different filters,
and then take their DNA back to our lab in Rockville,
where we can sequence a hundred million letters
of the genetic code every 24 hours.
And with doing this,
we've made some amazing discoveries.
For example, it was thought that the visual pigments
that are in our eyes -- there was only one or two organisms
in the environment that had these same pigments.
It turns out, almost every species
in the upper parts of the ocean
in warm parts of the world
have these same photoreceptors,
and use sunlight as the source of their energy
and communication.
From one site, from one barrel of seawater,
we discovered 1.3 million new genes
and as many as 50,000 new species.
We've extended this to the air
now with a grant from the Sloan Foundation.
We're measuring how many viruses and bacteria
all of us are breathing in and out every day,
particularly on airplanes
or closed auditoriums.
(Laughter)
We filter through some simple apparatuses;
we collect on the order of a billion microbes from just a day
filtering on top of a building in New York City.
And we're in the process of sequencing all that
at the present time.
Just on the data collection side,
just where we are through the Galapagos,
we're finding that almost every 200 miles,
we see tremendous diversity
in the samples in the ocean.
Some of these make logical sense,
in terms of different temperature gradients.
So this is a satellite photograph
based on temperatures -- red being warm,
blue being cold --
and we found there's a tremendous difference between
the warm water samples and the cold water samples,
in terms of abundant species.
The other thing that surprised us quite a bit
is these photoreceptors detect different wavelengths of light,
and we can predict that based on their amino acid sequence.
And these vary tremendously from region to region.
Maybe not surprisingly,
in the deep ocean, where it's mostly blue,
the photoreceptors tend to see blue light.
When there's a lot of chlorophyll around,
they see a lot of green light.
But they vary even more,
possibly moving towards infrared and ultraviolet
in the extremes.
Just to try and get an assessment
of what our gene repertoire was,
we assembled all the data --
including all of ours thus far from the expedition,
which represents more than half of all the gene data on the planet --
and it totaled around 29 million genes.
And we tried to put these into gene families
to see what these discoveries are:
Are we just discovering new members of known families,
or are we discovering new families?
And it turns out we have about 50,000
major gene families,
but every new sample we take in the environment
adds in a linear fashion to these new families.
So we're at the earliest stages of discovery
about basic genes,
components and life on this planet.
When we look at the so-called evolutionary tree,
we're up on the upper right-hand corner with the animals.
Of those roughly 29 million genes,
we only have around 24,000
in our genome.
And if you take all animals together,
we probably share less than 30,000
and probably maybe a dozen
or more thousand different gene families.
I view that these genes are now
not only the design components of evolution.
And we think in a gene-centric view --
maybe going back to Richard Dawkins' ideas --
than in a genome-centric view,
which are different constructs of these gene components.
Synthetic DNA, the ability to synthesize DNA,
has changed at sort of the same pace
that DNA sequencing has
over the last decade or two,
and is getting very rapid and very cheap.
Our first thought about synthetic genomics came
when we sequenced the second genome back in 1995,
and that from mycoplasma genitalium.
And we have really nice T-shirts that say,
you know, "I heart my genitalium."
This is actually just a microorganism.
But it has roughly 500 genes.
Haemophilus had 1,800 genes.
And we simply asked the question,
if one species needs 800, another 500,
is there a smaller set of genes
that might comprise a minimal operating system?
So we started doing transposon mutagenesis.
Transposons are just small pieces of DNA
that randomly insert in the genetic code.
And if they insert in the middle of the gene, they disrupt its function.
So we made a map of all the genes
that could take transposon insertions
and we called those "non-essential genes."
But it turns out the environment is very critical for this,
and you can only
define an essential or non-essential gene
based on exactly what's in the environment.
We also tried to take a more directly intellectual approach
with the genomes of 13 related organisms,
and we tried to compare all of those, to see what they had in common.
And we got these overlapping circles. And we found only 173 genes
common to all 13 organisms.
The pool expanded a little bit if we ignored
one intracellular parasite;
it expanded even more
when we looked at core sets of genes
of around 310 or so.
So we think that we can expand
or contract genomes, depending on your point of view here,
to maybe 300 to 400 genes
from the minimal of 500.
The only way to prove these ideas
was to construct an artificial chromosome with those genes in them,
and we had to do this in a cassette-based fashion.
We found that synthesizing accurate DNA
in large pieces was extremely difficult.
Ham Smith and Clyde Hutchison, my colleagues on this,
developed an exciting new method
that allowed us to synthesize a 5,000-base pair virus
in only a two-week period
that was 100 percent accurate,
in terms of its sequence and its biology.
It was a quite exciting experiment -- when we just took the synthetic piece of DNA,
injected it in the bacteria and all of a sudden,
that DNA started driving the production of the virus particles
that turned around and then killed the bacteria.
This was not the first synthetic virus --
a polio virus had been made a year before --
but it was only one ten-thousandth as active
and it took three years to do.
This is a cartoon of the structure of phi X 174.
This is a case where the software now builds its own hardware,
and that's the notions that we have with biology.
People immediately jump to concerns about biological warfare,
and I had recent testimony before a Senate committee,
and a special committee the U.S. government has set up
to review this area.
And I think it's important to keep reality in mind,
versus what happens with people's imaginations.
Basically, any virus that's been sequenced today --
that genome can be made.
And people immediately freak out about things about Ebola or smallpox,
but the DNA from this organism is not infective.
So even if somebody made the smallpox genome,
that DNA itself would not cause infections.
The real concern that security departments have
is designer viruses.
And there's only two countries, the U.S. and the former Soviet Union,
that had major efforts
on trying to create biological warfare agents.
If that research is truly discontinued,
there should be very little activity
on the know-how to make designer viruses in the future.
I think single-cell organisms are possible within two years.
And possibly eukaryotic cells,
those that we have,
are possible within a decade.
So we're now making several dozen different constructs,
because we can vary the cassettes and the genes
that go into this artificial chromosome.
The key is, how do you put all of the others?
We start with these fragments,
and then we have a homologous recombination system
that reassembles those into a chromosome.
This is derived from an organism, deinococcus radiodurans,
that can take three million rads of radiation and not be killed.
It reassembles its genome after this radiation burst
in about 12 to 24 hours,
after its chromosomes are literally blown apart.
This organism is ubiquitous on the planet,
and exists perhaps now
in outer space due to all our travel there.
This is a glass beaker after
about half a million rads of radiation.
The glass started to burn and crack,
while the microbes sitting in the bottom
just got happier and happier.
Here's an actual picture of what happens:
the top of this shows the genome
after 1.7 million rads of radiation.
The chromosome is literally blown apart.
And here's that same DNA automatically reassembled
24 hours later.
It's truly stunning that these organisms can do that,
and we probably have thousands,
if not tens of thousands, of different species
on this planet that are capable of doing that.
After these genomes are synthesized,
the first step is just transplanting them
into a cell without a genome.
So we think synthetic cells are going to have tremendous potential,
not only for understanding the basis of biology
but for hopefully environmental and society issues.
For example, from the third organism we sequenced,
Methanococcus jannaschii -- it lives in boiling water temperatures;
its energy source is hydrogen
and all its carbon comes from CO2 it captures back from the environment.
So we know lots of different pathways,
thousands of different organisms now
that live off of CO2,
and can capture that back.
So instead of using carbon from oil
for synthetic processes,
we have the chance of using carbon
and capturing it back from the atmosphere,
converting that into biopolymers
or other products.
We have one organism that lives off of carbon monoxide,
and we use as a reducing power
to split water to produce hydrogen and oxygen.
Also, there's numerous pathways
that can be engineered metabolizing methane.
And DuPont has a major program with Statoil in Norway
to capture and convert the methane
from the gas fields there into useful products.
Within a short while, I think there's going to be a new field
called "Combinatorial Genomics,"
because with these new synthesis capabilities,
these vast gene array repertoires
and the homologous recombination,
we think we can design a robot to make
maybe a million different chromosomes a day.
And therefore, as with all biology,
you get selection through screening,
whether you're screening for hydrogen production,
or chemical production, or just viability.
To understand the role of these genes
is going to be well within reach.
We're trying to modify photosynthesis
to produce hydrogen directly from sunlight.
Photosynthesis is modulated by oxygen,
and we have an oxygen-insensitive hydrogenase
that we think will totally change this process.
We're also combining cellulases,
the enzymes that break down complex sugars into simple sugars
and fermentation in the same cell
for producing ethanol.
Pharmaceutical production is already under way
in major laboratories
using microbes.
The chemistry from compounds in the environment
is orders of magnitude more complex
than our best chemists can produce.
I think future engineered species
could be the source of food,
hopefully a source of energy,
environmental remediation
and perhaps
replacing the petrochemical industry.
Let me just close with ethical and policy studies.
We delayed the start of our experiments in 1999
until we completed a year-and-a-half bioethical review
as to whether we should try and make an artificial species.
Every major religion participated in this.
It was actually a very strange study,
because the various religious leaders were using their scriptures as law books,
and they couldn't find anything in them prohibiting making life,
so it must be OK. The only ultimate concerns
were biological warfare aspects of this,
but gave us the go ahead to start these experiments
for the reasons we were doing them.
Right now the Sloan Foundation has just funded
a multi-institutional study on this,
to work out what the risk and benefits to society are,
and the rules that scientific teams such as my own
should be using in this area,
and we're trying to set good examples as we go forward.
These are complex issues.
Except for the threat of bio-terrorism,
they're very simple issues in terms of,
can we design things to produce clean energy,
perhaps revolutionizing
what developing countries can do
and provide through various simple processes.
Thank you very much.