The recombinant DNA
controversy
“Those who disregard the past are bound to repeat it”
George Santayana
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Herb Boyer
(EcoRI)
Stanley Cohen
(pSC101)
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J. Bacteriol. 64(4): 557–569 (1952)
1953
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http://opbs.okstate.edu/~melcher/MG/MGW3/MG331.html
Werner Arber (1965): met-depleted E. coli don’t generate “modified” phage
 CH3 of phage DNA!!
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The restriction/modification system:
a bacterial pathway for defence against viruses
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Dan Nathans
Ham Smith
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“How restriction enzymes became the
workhorses of molecular biology”
A significant breakthrough came in 1970 when the first of two papers
from Smith's laboratory described an enzyme, endonuclease R, that
was able to cleave bacteriophage T7 DNA into specific fragments
(2). This was the first type II restriction enzyme, the sort that now
populates our freezers, because it recognize specific sequences and
also gives rise to very specific cleavage. Smith had been looking for
an enzyme that might be involved in site-specific recombination in
Haemophilus influenzae and thought at first that endonuclease R
might be his long-sought quarry. With Tom Kelly, he went on to
determine the DNA sequence recognized by endonuclease R and
reported it as GTY RAC (11). This sequence seemed too short for a
recombination enzyme, and during correspondence with his close
friend Nathans, who ran the neighboring laboratory but was away on
sabbatical, it became clear that this enzyme might have very
practical uses for the analysis of DNA.
R. Roberts – PNAS 2005
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Radioautogram of 14C-labeled SV40 DNA cleaved with endonuclease R
showing 11 distinct fragments. Figure 3 from:
Danna, K. & Nathans, D. (1971) Proc. Natl. Acad. Sci. USA 68, 2913-2917
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plasmids
9.7
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9.12
1.8 million
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SV40
(primate polyomavirus)
1972:
E. coli
(resident of human GI tract)
+
=?
Michael Rogers “Biohazard” (1977)
The question, quite simply, concerned the wisdom of transplanting SV40 genes,
conceivably coding for tumor production, into a bacterium that not only lacks that
capacity to begin with, but which lives in virtually every human gut on the planet.
Berg: “ ‘My God! – people said – ‘You cannot put SV40 into E. coli! I
think I was upset by the criticism at first, but then I went out and
started to talk about the problem with a lot of people. … I realized
that I’d been wrong many, many times before in predicting the
outcome of an experiment, and that if I was wrong about my
assessment of the risk in this experiment, then the consequences
were not something that I would want to live with.”
Berg cancelled the experiment.
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Science, September 1973: letter to the NAS from participants at Gordon
conference on nucleic acids.
“We are writing … on behalf of a number of scientists to communicate a
matter of deep concern. .We presently have the technical ability to join
together, covalently, DNA molecules from diverse sources... This technique
could be used, for example, to combine DNA from animal viruses with
bacterial DNA... In this way, new kinds of hybrid plasmids or viruses, with
biological activity of unpredictable nature, may eventually be created. These
experiments offer exciting and interesting potential, both for advancing
knowledge of fundamental biological processes, and for alleviation of human
health problems. Certain such hybrid molecules may prove hazardous to
laboratory workers and to the public. Although no hazard has yet been
established, prudence suggests that the potential hazard be seriously
considered.”
Maxine Singer (Carnegie)
Dieter Söll (Yale)
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September 1974 – a moratorium?
New plasmids: novel antibiotic resistance
markers into E. coli; Xenopus and
Drosophila genomic DNA into E. coli.
Norton Zinder: “If we had any guts at all,
we’d tell people to not do these
experiments!”
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Sept. ’74 – Feb’ 75
“Until the potential hazards of such recombinant
DNA molecules have been better evaluated or
until adequate methods are developed for
preventing their spread, scientists throughout
the world join with the members of this
committee in voluntarily deferring the following
experiments” – cloning new antibiotic resistance
genes in currently naïve hosts and cloning
fragments of oncoviral genomes into bacterial
plasmids.
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“an epidemic of cancer”?
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Asilomar conference, February 1975
Paul Berg, David Baltimore, Sydney Brenner, Mike
Bishop, Don Brown, Ron Davis, James Watson,
Phil Sharp, Herb Boyer, Joshua Lederberg + 150
more people
+ Nature, Science, NY Times, Washington Post,
Boston Globe, SF Chronicle, etc
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(Left to right) Maxine Singer, Norton Zinder, Sydney Brenner, and Paul Berg
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The measures
1. Moratorium lifted.
2. Experiments will proceed under two levels of
containment, biological (use weakened host
cell), and physical, the level of which will be
gauged by the level of risk presented by the
organism created (minimal, low, moderate,
high):
1.
2.
3.
4.
Stringent, commonsense cleanliness (P1)
Similar to surgical operating theatre (P2)
Giant isolator (P3)
Comparable to what one uses in biological warfare
research (P4)
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Where to draw the cutoff line, idea #1
invertebrates
(including flies)
Stringent, commonsense cleanliness (P1)
vertebrates
Similar to surgical operating theatre (P2)
Giant isolator (P3)
Comparable to what one uses in biological warfare research (P4)
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Where to draw the cutoff line, idea #2
invertebrates
and cold-blooded
vertebrates
(including frogs)
Warm-blooded
vertebrates
Stringent, commonsense cleanliness (P1)
Similar to surgical operating theatre (P2)
Giant isolator (P3)
Comparable to what one uses in biological warfare research (P4)
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Enter the sceptics
Robert Sinsheimer (CalTech):
• The dangers of “shotgunning” –
evolutionary jumping? – creation of
chimeras with completely unforeseen new
properties?
• Prokaryote-to-eukatyote gene transfer?
• Use of E. coli as a host is potentially
dangerous
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Genomic
library:
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RNA splicing removes introns
• Exons – sequences found in a gene’s DNA
and mature mRNA (expressed regions)
• Introns – sequences found in DNA but not
in mRNA (intervening regions)
• Some eukaryotic genes have many introns
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Dystrophin gene underlying Duchenne muscular
dystrophy (DMD) is an extreme example of introns
Fig. 8.15
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Spliced Segments at the 5’ Terminus of Adenovirus 2 Late mRNA
Susan M. Berget, Claire Moore, and Phillip A. Sharp
Proceedings of the National Academy of Sciences USA, Volume 74, 1977,
pages 3171 3175
Also:
Louise Chow, Richard Gelinas, Tom Broker, and Richard Roberts
An amazing sequence arrangement at the 5 ends of adenovirus 2 messenger RNA,
Cell 12: 1 8, 1977
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How RNA processing splices out
introns and adjoins adjacent exons
Fig. 8.16
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• Splicing is
catalyzed by
spliceosomes
– Ribozymes –
RNA molecules
that act as
enzymes
– Ensures that all
splicing
reactions take
place in concert
Fig. 8.17
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“What seems to have been disregarded completely is that we
are dealing here much more with an ethical problem than one
in public health, and that the principal question to be
answered is whether we have the right to put an additional
fearful load on generations that are not yet born… You can
stop splitting the atom; you can stop visiting the moon… But
you cannot recall a new form of life. Once you have
constructed a viable E. coli cell carrying a plasmid DNA into
which a piece of eukaryotic DNA has been spliced, it will
survive you and your children and your children’s children.
An irreversible attack on the biosphere is something so
unheard of, so unthinkable to previous generations, that I
could only wish that mine had not been guilty of. The
hybridization of Prometheus with Herostratus is bound to give
evil results. … My generation … has been the first to engage,
under the leadership of exact sciences, in a destructive
colonial war against nature. The future will curse us for it.
Chargaff E, Simring FR. “On the dangers of genetic meddling.” Science. (1976) 192:938
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Enter the public, part 1
June 8, 1976: “Biohazards at Harvard:
scientists will create new life forms – but
how safe will they be?”
John Lear Recombinant DNA: the Untold Story
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“Frankenstein monsters from
Harvard Yard”
Mayor Alfred Vellucci: “They may come up with a disease that
can’t be cured – even a monster! Is this the answer to Dr.
Frankenstein’s dream?”
 an open meeting of the Cambridge City Council, June 13,
1976
“Why didn’t someone advise the mayor of Cambridge and the
CCC what was going on at Harvard?”
Resolution: ban all recombinant DNA research in the city,
whether publicly or privately financed, for two years.
 Establish Cambridge Experimentation Review Board. City
Manager Sullivan chose to name an ERB consisting entirely
of nonscientists.
John Lear Recombinant DNA: the Untold Story
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Not only Cambridge
City ordinances requiring uniform
compliance with city-specified controls on
working with recombinant DNA:
Princeton, Emeryville, Berkeley (Sept. 77),
Amherst, etc etc.
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Enter the public, part 2
The opening session of a three-day forum in
March 1977, sponsored by the National
Academy of Sciences, … was disrupted by a
citizens’ group, The People’s Business
Commission, led by activist Jeremy Rifkin,
whose members, chanting “We Will Not Be
Cloned,” draped across the platform a
banner with the caption, “We Will Create the
Perfect Race.”
S. Wright Molecular Politics
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The severe adverse events that have occurred in
the subsequent 30 years of nonstop cloning of all
imaginable sorts of DNA into E. coli vectors
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A more severe example of the
society-genetics interface
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Approximately 1.4 billion people, 25% of the world’s
population, receive insufficient food to meet daily
requirements.
Even with the heavy use of various pesticides,
approximately 50% of the world’s food crops are destroyed
in the field or in storage.
Levetin and McMahon Plants and Society
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After the revolution of 1917,
agriculture in Russia
collapsed. Formerly one of the
world’s greatest agricultural
nations became a shadow of a
shadow of its former self.
Something needed to be done.
Stalin decided that the way to go
was nationalize agriculture. In
1930, some 10 million landowning peasants were killed,
and their land was transferred
to the state.
Agricultural production levels
were still unacceptably low,
and hunger was ubiquitous.
Some 5 million people
perished in the Ukraine,
formerly an agricultural
paradise comparable to
California, in the Great Famine
of 1932-34.
Dmitry Mor Help – 1922
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Technical problem
Getting better yielding,
or more pestresistant, or more
cold-resistant crop
varieties by
conventional selection
takes TIME.
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Nikolai Koltsov
Genetics
Trofim Lysenko
Sergei Chetverikov
Population genetics
Nikolai Vavilov
Plant genetics and evolution
Iosif (Josef) Stalin
See the book “Commissar Vanishes” and also:
http://www.newseum.org/berlinwall/commissar_vanishes/
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The two central tenets of Lysenkoism
1.
2.
Lamarck was right. Acquired traits can be inherited.
From an agricultural perspective, this meant that plant
lines with desired characteristics could be obtained by
treating a set of plants a certain way, getting them to
develop a desired characteristic, and then breeding
them. It was then claimed that the F1 would have the
parents’ trait.
How could inheritance of acquired traits work?!
Weissman (“continuity of germ plasm”), Mendel
(particulate inheritance) and Morgan (genes lie on
chromosomes in the nucleus) are wrong. The entire
cell (or the organism) is a carrier of hereditary
information. “Gene” and “chromosome” are bourgeois,
capitalist inventions.
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Lysenko, 1936
“The most wonderful forms of animals and
plants are found in Nature. … Man will be
able to create similarly wonderful forms
within an immeasurably shorter time frame
… he will be able to create forms that have
never existed and could not have
appeared in Nature in a million years.”
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1948: Apocalypse Now
Annual Meeting of the Soviet Academy of Agricultural
Sciences:
Lysenko announces that “Comrade Stalin has not only read
my talk, but approves of it.”
Result: genetics banned and declared a pseudoscience
(“Weissmanism-Morganism-Mendelism”).
Geneticists are officially dubbed “fly lovers = people haters”
(мухолюбы – человеконенавистники).
Countless geneticists go through the same experience as,
earlier, Chetverikov (sent into exile, 1929) and Vavilov
(who died in a prison camp, 1943).
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Cheap populism and demagoguery:
a textbook example
“Genetics, mendelism-morganism, the fruitlesness of which
can be considered to be proven, has nothing to do with
our plan to harvest enough grain. … There is a science
[Lysenko’s “theories”] that is making a major contribution
to our [goals in agriculture].
The truth is that our people, our working class and our
peasants are decisively removing from science all that is
decrepit, all that is against the people, all that is borne
from a slave-like allegiance to the bourgeois West.
The truth is that [Lysenko’s “theories”] have been tested,
appreciated, confirmed and beloved by the masses, and
not only in [research institutes].”
A. Mikhalevich, speech at the 1948 agricultural academy annual meeting,
cited in Nikolai Dubinin, “The History and Tragedy of Soviet Genetics.”
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A perspective
The central phenomenon of Lysenkoism, and the enormity
of the nightmare that ensued, was a blatant invasion of
ideology into science.
The scientific method was abandonded, and notions were
declared “right” or “wrong” based on whether they fit a
certain ideology, not whether there are data supporting
or refuting them.
It did not matter to Stalin’s henchmen in science, what the
data showed. The only thing that mattered was whether
a certain theory fit the ideology of lysenkoism. Morgan’s
chromosome “theory” of inheritance, or Avery’s ‘theory”
that DNA carries genetic information were proclaimed as
wrong because they were the products of capitalist
ideology.
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Further reading
“Lysenko and the tragedy of Soviet science”
Valery Soyfer
“The rise and fall of T. D. Lysenko”
Zhores Medvedev
“The Lysenko affair”
David Joravsky
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A European Perspective on ID
It is a strange experience for a European biologist to read about the growing
support in the United States for so-called "intelligent design," the current name
for good old unintelligent creationism. Strangest of all, though, are the recent
activities of the Kansas Board of Education. The Kansas Board's proposal to
"[change] the definition of science" is unheard of in a western democracy,
although similar activities have been common in dictatorships. In Nazi
Germany, relativity was considered "Jewish science" and therefore
unacceptable, while in the Soviet Union, modern genetics was rejected as
unmarxist in favor of the ravings of the charlatan Lysenko. Is this the way the
good citizens of Kansas (and the many other states where similar initiatives are
seen) want to go? Obviously, there must be a profound ignorance of science
and the scientific method among the U.S. public for such a thing to happen (an
ignorance that intelligent design supporters evidently hope to perpetuate), and
for this, scientists must be held responsible. There is too much looking down at
colleagues who engage the public through popular science, such as the late
Carl Sagan (1). All scientists, not just biologists, should realize that an attack on
the very roots of science concerns every one of them, and accordingly, they
should do their utmost to counteract it by actively participating in the debate.
Ejnar J. FJERDINGSTAD Retired Professor of Anatomy, University of Aarhus,
Denmark.
Science July 29, 2005
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Wed/Fri
What if – if! – acquired characteristics could
be inherited?...
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