Biotechnology: Timeline 2005
Tuesday, August 30, 2005

Print out and Read these articles before class Tuesday: (PS. Please register for the New York Times - free)
(1) Nature, January 2003: The Birth of Biotechnology
(2) Scientific American, March 1997: Announcing Dolly
(3) From NHGRI (genome.gov), April 2005: Polymerase Chain Reaction (PCR) fact sheet
(4) Science, February 2001:The Human Genome: Barbara R. Jasny and Donald Kennedy
(5) New York Times, 12 February 2004: Human Embryos Created through Cloning
(6) Nature, 21 October 2004: Human genome: End of the beginning 431, 915 - 916
(7) Science,17 June 2005 Patient-Specific Embryonic Stem Cells Derived from Human SCNT Blastocyst Hwang et al. 308:1777-1783 [Note: A 7-page PDF, and also a pretty heavy-duty scientific paper! Please read Pages 1-2 only and we will read the rest later in the semester when we talk about cloning!

Steps on the Road to Biotechnology....

1859 Charles Darwin publishes On the Origin of Species, establishing the Theory of Evolution and its mechanism, natural selection, and challenging both the accepted scientific and religious views of Western culture that had been taught for centuries. The first edition sold out on the day it was published. Theologians quickly labeled Darwin "the most dangerous man in England"... Yet, after reading it, Darwin's friend and colleague T. H. Huxley had a different reaction: "How extremely stupid not to have thought of that." Quote

1865 The age of genetics begins when Gregor Mendel, studying inherited traits of pea plants, outlines the basic laws of heredity that still hold true today for all organisms. Mendel's discoveries about "heritable factors" (genes) are not recognized by other scientists for over 35 years.

1910 Chromosomal theory of inheritance proposed: Thomas Hunt Morgan establishes that genes are located on chromosomes by physically tracing a specific gene to a specific chromosome. Morgan wins the 1933 Nobel Prize in Medicine.

1941 One gene, one enzyme: George Beadle and Edward Tatum establish that one gene makes one enzyme or protein, and share the 1958 Nobel Prize in Medicine.

1952 The "Waring Blender Experiment": Martha Chase and Alfred Hershey use the common kitchen appliance to separate the protein coats of viruses from their DNA to demonstrate that DNA is the substance that transmits inherited characteristics from one generation to the next. Hershey wins the 1969 Nobel Prize.

1953 Unraveling the double helix. James Watson and Francis Crick deduce the structure of the DNA molecule - a double helix - without ever doing a single experiment at the bench. In a classic "race to the finish", Watson and Crick submit a one-page paper to the journal Nature, starting with "We wish to suggest a structure for the salt of deoxyribose nucleic acid (D.N.A.)", and ending with the subtle understatement: "It has not escaped our notice that the specific pairing that we have postulated immediately suggests a possible copying mechanism for the genetic material". Their work is recognized with the 1962 Nobel Prize, shared with Maurice Wilkins (Rosalind Franklin died 4 years earlier) [Image]

1967 Cracking the the genetic code. Har Khorana, Robert Holley, and Marshall Nirenberg decipher the mechanism that enables DNA to be translated into proteins. Nirenberg, Khorana, and Holley share the 1968 Nobel Prize.

1968 Stanley Cohen, studying bacterial disease at Stanford, determines that bacteria carry genes for antibiotic resistance on plasmids, extrachromosomal circles of DNA. Cohen learns how to purify plasmids and reinsert them into other bacterial cells, transferring antibiotic resistance in the process. [Image]

1970 Restriction enzymes discovered. UCSF scientist Herb Boyer, working with bacteriophages, discovers that certain bacteria preferentially fought off (or "restricted") certain phages by producing enzymes that chopped up the phage's DNA, leaving "sticky ends" on the cut strands. Boyer isolates the "Big Daddy" of restriction enzymes, EcoR1. In the ensuing years, hundreds of different restriction endonucleases are found that cleave DNA at specific sites. Earlier investigators studying restriction enzymes (Hamilton Smith et al.) win the1978 Nobel Prize in Medicine. [Image]

1972 Recombinant DNA technology begins: Stanford biochemist Paul Berg splices together two blunt-ended fragments of DNA from the SV40 virus and E. coli, creating recombinant DNA. Berg shares the 1980 Nobel Prize in Chemistry "for his fundamental studies of the biochemistry of nucleic acids, with particular regard to recombinant-DNA" (with Walter Gilbert and Fred Sanger below). [Image]

1972 "From Corned Beef to Cloning": In November, at a scientific meeting in Hawaii, Cohen hears Boyer describe his work with EcoR1 and his findings that the sticky ends of DNA can be linked together or "spliced" with DNA ligases. Cohen and Boyer meet at a Waikiki Beach Deli, where they discuss ways to combine plasmid isolation with DNA splicing. They form the idea of inserting desired DNA into bacterial plasmds that would then churn out specific proteins - the basis of the biotechnology industry. Quote

1975 Asilomar Conference: Pacific Grove CA: Paul Berg organizes an international conference on recombinant DNA technology with over 100 other scientists to discuss what they knew (and didn't know) about recombinant DNA and to draw up guidelines that would let the science proceed without undue risk. The scientists agree to suspend research involving recombinant DNA technology research until potential risks can be evaluated, a "milestone of self-regulation in science". Although recombinant DNA technology turned out to be much more harmless than many had suspected, Asilomar remains an important scientific landmark, a rare instance of scientists independently questioning and successfully regulating their own work. Quote

1975 DNA sequencing developed: Walter Gilbert and Allan Maxam of Harvard University and Fred Sanger of Cambridge University simultaneously come up with two techniques for determining the exact sequence of bases that make up a gene. Gilbert and Sanger share the 1980 Nobel Prize (with Paul Berg).

1975 Cesar Milstein, Georges Kohler and Niels Jeme develop monoclonal antibody technology by fusing immortal tumor cells with antibody-producing B lymphocyte cells to produce "hybridomas," that continuously synthesize identical (or "monoclonal") antibodies. Milstein, Kohler and Jeme are awarded the 1984 Nobel Prize in Medicine.

1976 Big bucks for Biotech: The commercial potential of using cells as factories for hormones and proteins to produce "biopharmaceuticals" is not lost on the business world. Robert Swanson, a 29-year-old Silicon Valley venture capitalist, and Herb Boyer team up to form Genentech, Inc. (for "GENetic ENgineering TECHnology") with the goal of cloning human insulin. Genentech goes public on Oct. 14, 1980, offering one million shares of stock for $35 a share - and makes $35 million in an afternoon. By the end of the day, Genentech's stock makes market history by hitting a high of $89, a record for an initial public offering. [Image] and Q .

1978 Human insulin cloned into E. coli by Genentech sciensists. Genentech licenses the human insulin technology to Eli Lilly. In 1982, human insulin, or Humulin, becomes the first recombinant DNA drug approved by FDA. [Image]

1985 Genentech becomes the first biotechnology company to launch its own biopharmaceutical product in 1985, ProTropin - growth hormone for children with growth hormone deficiency.

1986 The Polymerase Chain Reaction (PCR) is conceived by Kary Mullis and revolutionizes molecular biology... PCR uses a thermostable DNA polymerase to amplify any given DNA segment billions of times in a few hours. Taq polymerase is chosen as the 1989 Molecule of the Year by the journal Science. Kary Mullis, having parted ways with his employer (Cetus) as well as alienated himself from the scientific establishment (we won't go there), is not even mentioned in the Science article. However, Mullis IS awarded the 1993 Nobel Prize in Chemistry and goes on to become a best-selling novelist. A long, bitter "David vs. Goliath" patent struggle between Hoffman-LaRoche and Promega Biotech over ownership of PCR technology and the key enzyme,Taq polymerase, was at last resolved in 1999 (in favor of Promega). [Image]

1989 The Human Genome Project (HGP) begins. An ambitious plan to "map, sequence and render accessible for further biological study" all ~100,000 human genes by the year 2005, initially led by director James Watson. Human chromosomes are parceled out to labs around the world, with new genomic sequencing technologies springing up to meet the need for faster sequence analysis. Anticipated cost: $3 billion (NIH / DOE). [Great timeline of its own: Exploring our Molecular Selves]

1990 First use of gene therapy to treat human patient. Ashanti DiSilva, a 4-year-old girl with ADA deficiency is the first recipient of gene therapy. William French Anderson and colleagues at the NIH insert a normal ADA gene into the girl's T-cells and re-introduce into her bloodstream. Injections of corrected T-cells every 2 months restores 25% of her immune system function, allowing her and others with ADA deficiency to lead a normal life.

1994 Brave new foods ­ On May 18, the Food and Drug Administration announces the arrival of Calgene's FlavrSavr tomato, the first transgenic food, to the supermarket shelves. FlavrSavr had undergone a decade of testing, costing $525 million, before being approved safe by the FDA. Engineered to remain firm even as it turns red and ripe, FlavrSavr "provides summertime taste year round". Although delicious, the FlavrSavr suffers from consumer resistance, high price and a boycott by chefs. Calgene, heavily in debt before the tomato hit the market, declares the FlavrSavr dead on the vine in 1997. [Image]

1996 First mammal cloned from adult cells: A surrogate mother sheep gives birth to Dolly, a lamb cloned from an udder cell of an adult sheep born 6 years earlier. Ian Wilmut and colleagues at the PPL Theraputics and the Roslin Institute in Scotland quietly announce the birth of Dolly in February, 1997 in the journal Nature, after which, all hell breaks loose..Sadly, on February 14, 2003: "Dolly the sheep, the first cloned mammal, is dead at age 6 of a lung infection common to sheep raised in barns. (snif!). .[Image]

1996 Development of the GeneChip®: The Department of Biochemistry at Stanford and Affymetrix introduce a technological breakthrough in gene expression and DNA sequencing technology with the introduction of DNA chips, small glass or silica microchips that contain thousands of individual genes that can be analyzed simultaneously. Since then, DNA Chip technology has become a growth industry as new tools for making, probing, imaging, and analyzing arrays are introduced almost daily. [Image]

1997 Three Cloned Mice: Dolly is joined on October 3, 1997 by the cloned mouse "Cumulina" and, shortly afterward, by 22 of her cloned siblings (some of whom were cloned from clones) using the 'Honolulu Technique' of nuclear transfer. Author Teruhiko Wakayama concludes that "contrary to previous opinion, mammals can be reproducibly cloned from adult somatic cells".

1997 First Human Artificial Chromosome: Scientists at Athersys in Cleveland OH use a combination of natural and synthetic DNA to create a "genetic cassette" that can potentially be customized and used in gene therapy. Genes on the artificial chromosome are expressed and replicated in cells for over 6 months. [Image]

1998 (May) Race for the Genome: J. Craig Venter and Perkin Elmer merge to create Celera Genomics. Goal: sequence the entire human genome by December 31, 2001 - 2 years before the completion by the HGP, and for a mere $300 million. The company is massive genomics sequencing facility with a capacity greater than that of the current combined world output, and second only to the Pentagon in computing power. Venter calls the plan a "mutually rewarding partnership between public and private institutions." Press release

1998 (October) Human Genome Project on 'Fast Track' for Early Completion: The DOE and NIH approve new 5-year goals aimed at completing the Human Genome Project in 2003, generating a "working draft" of the human genome DNA sequence by 2001. PS. "It's NOT a race"

1998 (November): Two research teams, led by James Thompson (UW Madison) John Gearhart (Johns Hopkins) succeed in growing human Embryonic Stem (hES) cells, pleuripotent, self renewing cells with the demonstrated ability to differentiate in vitro into all three embryonic germ layers. Science selected stem cell research and technology as the 1999 "Breakthrough of the Year." The research was funded by and is licensed to Geron Corporation.

1999 (September) Speed Matters: Celera announces completion of the Drosophila genome sequence (With Gerry Rubin et. al. of Howard Hughes Medical Institute) on September 9, and immediately begins sequencing the Human genome. Critics had predicted that (a) the full sequence would not be able to be deciphered and (b) that Celera would not release the sequence to the public, neither of which proved to be the case. Science reports the full scoop. [Image]

14 March 2000: Code Red for Biotech Stocks: President Clinton and Prime Minister Tony Blair released a joint statement that genome information "should be made freely available to scientists everywhere". While Clinton and Blair went on to reinforce “the intellectual property protection for all gene-based inventions”, the market seemed to react to only the first part of the statement, putting stock market investors in a panic. Biotech stocks across the board went into a 'screaming nosedive', dragging down the NASDAQ, which on that day suffered its second-biggest point loss ever! By the end of the day, investors in the biotechnology sector lost over $40 billion. Ouch! Image

26 June 2000 - The Race is Over: President Clinton, Tony Blair, the HGP, and Celera announce the completion of a "working draft" sequence of the human genome. The achievement provides scientists with a road map to the location and sequence of an estimated 90% of genes on every chromosome, with all HGP data freely available on the Internet. Although the draft contains gaps and errors, it provides a high-quality reference genome sequence -- with the final draft expected by 2003 or sooner. Quote (PS. Celera made the announcement to the White House on April 6, 2000, but decides to make a joint announcement with the HGP). [Image] Contest: Provide a caption for this photo...

26 January  2001: Inaugural Indecision: U.S. President George W. Bush, 43rd President of the United States, is undedcided whether to allow federal funding for stem cell research, despite letters of support from Nobel laureates, support from both conservative and liberal Senators, and a lawsuit by Christopher Reeves and concerned scientists.

February 15 and 16, 2001: The simultaneous publication of the historic Genome Issues of Nature (Collins et. al, Human Genome Project) and Science (Venter et. al, Celera Genomics). Both feature free full-text access, so check them out!

The Biggest Surprise about the Human Genome: The human genome contains only about 35,000 genes, just a fraction more than many 'lower' organisms and far fewer than numbers originally predicted for humans.

31 July 2001: Cloning Ban: by a vote of 265 to 162, the US House of Representatives passes the Human Cloning Prohibition Act of 2001 (HR2502), a ban on all human cloning, either for reproduction or for therapeutic cloning to derive immunologically compatible embryonic stem cells. Despite warnings, on August 7th, at a National Academy of Sciences Meeting, 3 privately-funded US / European groups vow to contunue their programs on Human Cloning - Dr. Brigitte Boisselier, Dr. Panos Zavos of Lexington KY, and Dr. Severino Antinori of Italy. (Note: The bill stalled in the Senate in June 2002).

10 August 2001: The Bush Decision on Stem Cells: In his first address to the nation, Bush approves a comprimise on stem cell funding. His decision allows for (a) full federal funding for research on adult and umbilical stem cells, (b) limited federal funding for research on human embryonic stem cells (hES cells) to pre-existing cell lines drawn from surplus embryos created for in-vitro fertilization, (c) no federal funding for research on hES cells from Donor Embryos created specifically for developing stem cells or for research in theraputic cloning (to obtain hES cells stem cells, tissues or organs that are genetically identical, and immunologically compatible, to the donor's). ["W", deep in the heart of Texas]

September 11, 2001...

23 January 2002: So it WAS his genome...! J. Craig Venter steps down as president of Celera Genomics, as Celera decides re-organize as a pharmaceutical business and drug discovery company. Venter announces the creation of the not-for-profit J. Craig Venter Science Foundation, initiated by $100 M of his own money. In his spare time Venter says he plans write a book "examining my own genetic code", revealing that he was one of the six supposedly anonymous donors used to generate Celera's genome sequence. [Quote]

15 August 2002: Next-generation genomes. The Institute for Genomic Research (TIGR) announces the formation of two non-profit organizations: the Institute for Biological Energy Alternatives (IBEA), analyzing genomes of organisms that metabolize carbon or hydrogen for cleaner energy alternatives, and The Center for the Advancement of Genomics (TCAG), a bioethics think-tank, supported by the J. Craig Venter Science Foundation. "Our goal is to build a new and unique sequencing facility that can deal with the large number of organisms to be sequenced, and can further analyze those genomes already completed," said Venter, "and at such reduced cost that health care customized to one's own DNA would be feasible". [Image] and [quotes].

28 January 2003: In his State of the Union Address, President Bush states to congress that "...because no human life should be started or ended as the object of an experiment, I ask you to set a high standard for humanity and pass a law against all human cloning."

28 February 2003: House Passes Ban on All Human Cloning: "The House bill bans all human cloning — for reproduction or research — and imposes a $1 million fine and a prison sentence of up to 10 years for violators." Competing bills have yet to be brought before the US Senate (stay tuned)
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14 April 2003: The Human Genome Project - fini! BETHESDA, Md., – "The International Human Genome Sequencing Consortium, led in the United States by the National Human Genome Research Institute (NHGRI) and the Department of Energy (DOE), today announced the successful completion of the Human Genome Project more than two years ahead of schedule. 'The Human Genome Project has been an amazing adventure into ourselves, to understand our own DNA instruction book, the shared inheritance of all humankind,' said NHGRI Director Francis S. Collins, M.D., Ph.D., leader of the Human Genome Project since 1993. "All of the project’s goals have been completed successfully – well in advance of the original deadline and for a cost substantially less than the original estimates." ...and just in time for.....

25 April 2003: Happy Birthday Double Helix! It's DNA Day! On April 25, 1953, James Watson and Francis Crick published their landmark letter to Nature describing the DNA double helix. Nature marks the 50th anniversary of the event with a free Nature web focus "containing news, features and web specials celebrating the historical, scientific and cultural impacts of the discovery of the double helix."

29 May 2003: Way to go, Idaho!: A healthy mule named Idaho Gem is the first member of the horse family to be cloned, by Gordon Wood et al. at the University of Idaho. Since mules can't have babies the good old fashioned way, cloning may allow breeders to produce identical copies of champion mules. Idaho Gem is the brother of Taz, a champion racing mule, and the Idaho Gem will also be trained to race. A second mule clone, Utah Pioneer, was born June 9th. Yee-hah! [Image]

7 August 2003: World's first cloned horse born to its genetic twin: LONDON, England (Reuters) -- "Italian scientists have created the world's first cloned horse from an adult cell taken from the horse who gave birth to her. Prometea weighed in at 80 lbs after a normal, full-term pregnancy. Although sheep, mice, cats, cattle, goats and pigs have already been cloned, Prometea is the first animal known to be carried and born by the mother from which she was cloned...". [Image]

10 December 2003: Chimp Genome Assembled the most closely related species to humans The sequence of the chimpanzee, Pan troglodytes, was assembled by NHGRI-funded teams led by Eric Lander, Ph.D., at The Eli & Edythe L. Broad Institute of the Massachusetts Institute of Technology and Harvard University, Cambridge, Mass.; and Richard K. Wilson, Ph.D., at the Genome Sequencing Center, Washington University School of Medicine, Saint Louis.

12 February 2004: Cloning Creates Human Embryos "Scientists in South Korea report he first human embryonic stem cell line produced with somatic cell nuclear transfer (cloning). Their goal, the scientists say, is not to clone humans but to advance understanding of the causes and treatment of disease. Patients with diseases like Parkinson's and diabetes have been waiting for the start of so-called therapeutic cloning to make embryonic stem cells that are an exact genetic match of the patient. Then those cells, patients hope, could be turned into replacement tissue to treat or cure their disease without provoking rejection from the body's immune system". We will talk about Human embryonic stem cells in Human Clone Produces Stem Cells 

26 February 2004: FDA Approves Avastin, the first Anti-Angiogenesis drug for treating cancer  "Genentech today announced the FDA approval of Avastin - the first FDA-approved therapy designed to inhibit angiogenesis, the process by which new blood vessels develop, which is necessary to support tumor growth and metastasis." Watch out, cancer cells! (Note: On August 13th, the FDA and Genetech released a warning that Avastin can increase the risk of clots that could cause a stroke or heart attack. Genentech shares fell nearly 6 percent on news of the warning (stay tuned).

2 April 2004: Sailing the Genome Seas: The Sorcerer II Expedition: J. Craig Venter, Ph.D., president of the Institute for Biological Energy Alternatives (IBEA), announced today in the journal Science (Environmental Genome Shotgun Sequencing of the Sargasso Sea, 2 April 2004) results from sequencing and analysis of samples taken from the Sargasso Sea off Bermuda. Using the whole genome shotgun sequencing and high performance computing developed to sequence the human genome, IBEA researchers sequenced over 1 billion bp of DNA, discovered at least 1,800 new species (mostly microbial) and more than 1.2 million new genes from the Sargasso Sea, all while sailing on Venter's 55-foot yacht .Check out the full story here! [Image]

14 July 2004: Woof! Dog Genome now available: "A team of scientists (MIT, Harvard, and Agencourt Bioscience) successfully assembled the genome of the domestic dog (Canis familiaris). The breed of dog was the boxer, one of the breeds with the least amount of variation in its genome and therefore likely to provide the most reliable reference genome sequence. Next mammals up: the orangutan, African elephant, shrew, the European hedgehog, the guinea pig, the lesser hedgehog , the nine-banded armadillo, the rabbit. and the domestic cat (each represents an important position on the mammalian evolutionary tree and is likely to by important in helping to interpret the human genome.)"

30 July 04: Francis Crick, DNA pioneer, dies at age 88 "Scientists around the world have paid tribute to British scientist Francis Crick, co-discover of the structure of DNA..."

12 August 2004: Green light for stem cell clones: Newcastle University (Britain) is granted first U.K. licence to create to create embryonic stem cells from human embryos for research. The decision adds the U.K. (with Korea) to the forefront of global research in hES cell technology.

17 June 2005 Patient-Specific Embryonic Stem Cells Derived from Human SCNT Blastocysts Hwang et al. Science, Vol 308, 1777-1783, (published online May 19) - 11 new “genetically matched ES cell lines for patients needing a cell transplant. It's a breakthrough that I didn't think would happen for decades..”

21 October 21, 2004, The Finished Sequence for Euchromatin - IHGSC, in Nature, announced a new refinement: "Finishing of the euchromatic sequence of the human genome" 2,851,330,913 nucleotides finished, at an error rate of 99.99% (no more than 1 error in 100,000). 341 gaps remain, (~228,000,000 nucleotides, ~198,000,000 bases of which is highly condensed heterochromatin - chromosomal replication and maintainence, only ~28,000,000 bases of active, gene-containing euchromatin). These gaps will have to await the technology to sequence them - not here yet!! A startling downward revision: Human Gene number revised (down) to only 20,000-25,000

29 July 2005: Frist stem cell support boosts biotechs Biotech analysts urge caution, as stock prices rocket on Frist's support. NEW YORK (CNN/Money) - Biotech stock prices surged after Senate Majority Leader Bill Frist stated his support for a bill that would provide federal funding for stem cell research, but analysts urged investor caution even as they hailed the good news. JDRF Hails Decision by Senator Frist to Support Stem Cell Research

3 August 2005: Announcing Snuppy: The first cloned puppy, "Seoul National University Puppy" The painstaking work was completed by - AGAIN - the lab of Woo Suk Hwang, the South Korean researcher who is famous for creating a cloned human embryo and deriving stem cells. Dogs cloned from adult somatic cells

24 August 2005 Lance Armstrong and Epo: Oui or Non??? French newspaper L'Equipereports that urine samples provided by Lance Armstrong during the '99 Tour de France tested positive for the red blood cell-booster EPO. The Question du Jour:"Les révélations sur le Tour 1999 de Lance Armstrong jettent-elles le discrédit sur l'ensemble de sa carrière?" :)

Objectives specific to this lecture

1. Identify Nobel-winning scientists and their research stated above.
2. Explain how the Cohen-Boyer partnership resulted in 'the birth of biotech', and the role of Robert Swanson developing the biotechnology company Genentech.
3. What was the Asilomar conference and what was its significance in the use of recombinant DNA technology?

In later discussions, we will focus in on (much) more specific objectives for the Human Genome Project, Cloning Animals, Stem Cells and Stem Cell legislation, Cloning Human Cells, Genomics, etc. You will want to keep returning to this timeline to help you study for those particular lectures!