TOPICS IN (NANO) BIOTECHNOLOGY Lecture I

1. PhD Course TOPICS IN (NANO) BIOTECHNOLOGY Lecture I 25th February, 2004

3. Overview • 1. What is Biotechnology? • Definitions of Biotechnology • Timeline of Biotechnology • Techniques used in Biotechnology • Who's Who in Biotechnology • 2. How is Biotechnology being used? • Applications of Biotechnology • Medicines on the market today • Agriculture - GM Foods and Animals • DNA fingerprinting and forensic science • Gene Therapy and Transgenic Animals • Human Embryonic Stem Cells and Cloning • 3. What are some of the societal issues Biotechnology raises? • Bioethics / "Genethics" • Public attitudes to biotechnology - safety, awareness • Therapeutic uses of human genes and tissues

4. What is biotechnology? • Biotechnology = bios (life) + logos (study of or essence) • Literally ‘the study of tools from living things’ • CLASSIC: The word "biotechnology" was first used in 1917 to describe processes using living organisms to make a product or run a process, such as industrial fermentations. (Robert Bud, The Uses of Life: A History of Biotechnology) • LAYMAN: Biotechnology began when humans began to plant their own crops, domesticate animals, ferment juice into wine, make cheese, and leaven bread (AccesExcellence)

5. What is biotechnology? • GENENTECH: Biotechnology is the process of harnessing 'nature's own' biochemical tools to make possible new products and processes and provide solutions to society's ills (G. Kirk Raab, Former President and CEO of Genentech) • WEBSTER’S: The aspect of technology concerned with the application of living organisms to meet the needs and ends of man. • WALL STREET:Biotechnology is the application of genetic engineering and DNA technology to produce therapeutic and medical diagnostic products andprocesses. Biotech companies have one thing in common - the use of genetic engineering and manipulation of organisms at a molecular level.

6. What is biotechnology? • Using scientific methods with organisms to produce new products or new forms of organisms • Any technique that uses living organisms or substances from those organisms or substances from those organisms to make or modify a product, to improve plants or animals, or to develop microorganisms for specific uses

7. What is biotechnology? • Biotechnology is a multidisciplinarian in nature, involving input from • Engineering • Computer Science • Cell and Molecular Biology • Microbiology • Genetics • Physiology • Biochemistry • Immunology • Virology • Recombinant DNA Technology  Genetic manipulation of bacteria, viruses, fungi, plants and animals, often for the development of specific products

8. What are the stages of biotechnology? • Ancient Biotechnology • early history as related to food and shelter, including domestication • Classical Biotechnology • built on ancient biotechnology • fermentation promoted food production • medicine • Modern Biotechnology • manipulates genetic information in organism • genetic engineering

9. Ancient biotechnology History of domestication and agriculture • Paleolithic society – Hunter-gatherers  Nomadic lifestyle due to migratory animals and edible plant distribution (wild wheat and barley) (~2 x 106 yrs.) • Followed by domestication of plants and animals (artificial selection)  People settled, sedentary lifestyles evolved (~10,000 yrs. ago) • Cultivation of wheat, barley and rye (seed collections) • Sheep and goats  milk, cheese, button and meat • Grinding stones for food preparation • New technology  Origins of Biotechnology  Agrarian Societies • History of domestication and agriculture History of domestication and agriculture History of domestication and agriculture

10. Ancient biotechnology Fermented foods and beverages • Long history of fermented foods since people began to settle (9000 BC) (fervere –to boil) • Often discovered by accident! • Improved flavor and texture • Deliberate contamination with bacteria or fungi (molds) • Examples: • Bread • Yogurt • Sour cream • Cheese • Wine • Beer • Sauerkraut

11. Ancient biotechnology Fermented foods and beverages • Dough not baked immediately would undergo spontaneous fermentation  would rise  Eureka!! • Uncooked fermented dough could be used to ferment a new batch  no longer reliant on “chance fermentation” • 1866 – Louis Pasteur published his findings on the direct link between yeast and sugars  CO2 + ethanol (anaerobic process) • 1915 – Production of baker’s yeast – Saccharomyces cerevisiae

12. Classical biotechnology Industry today exploits early discoveries of the fermentation process for production of huge numbers of products • Different types of beer • Vinegar • Glycerol • Acetone • Butanol • Lactic acid • Citric acid • Antibiotics – WWII (Bioreactor developed for large scale production, e.g. penicilin made by fermentation of penicillium) • Today many different antibiotics are produced by microorganisms • Cephalosporins, bacitracin, neomycin, tetracycline……..)

13. Classical biotechnology Chemical transformations to produce therapeutic products • Substrate  + Microbial Enzyme  Product • Examples: • Cholesterol  Steroids (cortisone, estrogen, progesterone) (hydroxylation reaction  -OH group added to cholesterol ring)

14. Classical biotechnology Microbial synthesis of other commercially valuable products • Amino acids to improve food taste, quality or preservation • Enzymes (cellulase, collagenase, diastase, glucose isomerase, invertase, lipase, pectinase, protease) • Vitamins • Pigments

15. Modern biotechnology • Cell biology • Structure, organization and reproduction • Biochemistry • Synthesis of organic compounds • Cell extracts for fermentation (enzymes versus whole cells) • Genetics • Resurrection of Gregor Mendel’s findings  1866  1900s • Theory of Inheritance (ratios dependent on traits of parents) • Theory of Transmission factors • W.H. Sutton – 1902 • Chromosomes = inheritance factors • T.H. Morgan – Drosophila melanogaster

16. Modern biotechnology • Molecular Biology • Beadle and Tatum (Neurospora crassa) • One gene, one enzyme hypothesis • Charles Yanofsky  colinearity between mutations in genes and amino acid sequence (E. coli) • Genes determine structure of proteins • Hershey and Chase – 1952 • T2 bacteriophage – 32P DNA, not 35S protein is the material that encodes genetic information

17. Modern biotechnology • Watson, Crick, Franklin and Wilkins (1953) • X-ray crystallography • 1962 – Nobel Prize awarded to three men • Chargaff – DNA base ratios • Structural model of DNA developed • DNA Revolution – Promise and Controversy!!! • Scientific foundation of modern biotechnology • based on knowledge of DNA, its replication, repair and use of enzymes to carry out in vitro splicing DNA fragments

18. Modern biotechnology • Breaking the Genetic Code – Finding the Central Dogma • An “RNA Club” organized by George Gamow (1954) assembled to determine the role of RNA in protein synthesis • Vernon Ingram’s research on sickle cell anemia (1956) tied together inheritable diseases with protein structure • Link made between amino acids and DNA • Radioactive tagging experiments demonstrate intermediate between DNA and protein = RNA • RNA movement tracked from nucleus to cytoplasm  site of protein synthesis

19. Modern biotechnology • DNA  RNA  Protein • TranscriptionTranslation • Genetic code determined for all 20 amino acids by Marshal Nirenberg and Heinrich Matthaei and Gobind Khorana – Nobel Prize – 1968 • 3 base sequence = codon

20. What are the areas of biotechnology? • Organismic biotechnology • uses intact organisms and does not alter genetic material • Molecular Biotechnology • alters genetic makeup to achieve specific goals • Transgenic organism: an organism with artificially altered genetic material

21. What are the benefits of biotechnology? • Medicine • human • veterinary • biopharming • Environment • Agriculture • Food products • Industry and manufacturing

22. What are the applications of biotechnology? • Production of new and improved crops/foods, industrial chemicals, pharmaceuticals and livestock • Diagnostics for detecting genetic diseases • Gene therapy (e.g. ADA, CF) • Vaccine development (recombinant vaccines) • Environmental restoration • Protection of endangered species • Conservation biology • Bioremediation • Forensic applications • Food processing (cheese, beer)

23. Anti-cancer drugs Transfer of new genes into animal organisms Culture of plants from single cells Diagnostics Crime solving Tracers DNA technology Genetic Engineering Synthesis of specific DNA probes Banks of DNA, RNA and proteins Cloning Synthesis of new proteins Mass prodn. of human proteins Localisation of genetic disorders Resource bank for rare human chemicals New types of plants and animals New antibiotics New types of food Gene therapy Cell Culture Monoclonal Antibodies Molecular Biology Complete map of the human genome

24. Biotechnology Timeline 1675 Leeuwenhoek discovers cells (bacteria, red blood cells) 1750 BC The Sumerians brew beer. 500 BC Chinese use moldy soybean curds as an antibiotic to treat boils 1590 Janssen invents the microscope 1830 Proteins are discovered 1833 The first enzymes are isolated 1855 The Eschirium coli bacterium is discovered

25. Biotechnology Timeline 1859 Charles Darwin publishes On the Origin of Species 1865 The age of genetics begins 1864 Louis Pasteur shows all living things are produced by other living things 1902 Walter Sutton coins the term ‘gene’ - proposed that chromosomes carry genes

26. Biotechnology Timeline 1910 Chromosomal theory of inheritance proved 1928 Fleming discovers antibiotic properties of certain molds 1941 George Beadle and Edward Tatum propose that one gene makes one protein 1949 Sickle cell anaemia demonstrated to be molecular disease

27. Biotechnology Timeline 1953 The double helix is unravelled 1973 Recombinant DNA technology begins 1952 The ‘Waring Blender’ experiment 1967 The genetic code is cracked 1975 First international conference on recombinant DNA technology

28. Biotechnology Timeline 1978 Kary Mullis discovers PCR 1975 DNA sequencing discovered 1975 Monoclonal antibody technology introduced 1978 Genentech Inc. established 1978 Genentech use genetic engineering to produce human insulin in E.coli - 1980 IPO of $89

29. Biotechnology Timeline 1994 The FLAVRSAVR tomato - first genetically engineered whole food 1989 The Human Genome Project begins 1990 First use of gene therapy 1990 First product of recombinant DNA technology introduced into US food chain 1993 FDA announces that transgenic food is safe

30. Biotechnology Timeline 1996 First mammal cloned from adult cells 1996 Development of Affymetrix GeneChip 1997 First artificial chromosome 1990s First conviction using genetic fingerprinting

31. History of Biotechnology 1999 Celera announces completion of Drosophilia genome sequence 2000 90% of Human Genome sequence published on web 2001 Human genome project complete 1998 Human embryonic stem cells grown

32. Excercise • Choose 3 scientists who have contributed to the biotechnology revolution and write a paragraph describing their input • Discover more about the Asilomar conference and decribe its signficance on the use of recombinant DNA technology • Discover more about what led to the death of the FLAVRSAVR tomato. • Follow one ‘linked set of discoveries’ outlining the path from the first experiment to today.

33. Excercise • Compare and argue both sides of Monsanto vs Greenpeace on the theme of genetically modified food • Outline the IMCLONE story and explain the potential impact on biotechnology industry. • Argue both for and against the use of human embryonic stem cells and outline international stance on this research. • Outline the story of the race to unravel DNA.

34. Discussion • What is the societal impression of biotechnology? • What are the negative impacts that biotechnology may have? • What are the potential ethical issues associated with biotechnology? • Why are biotechnology companies targeted by anti-globalisation protesters? • How can the image of biotechnology to the public be improved? Should it be improved? • What are the potential dangers of biotechnology?

35. Useful Resources • http://www.geocities.com/cwfennhcc/bi200/intro.html • http://www.geocities.com/cwfennhcc/bi200/quiz1.htm • http://www.accessexcellence.org/ • The Uses Of Life – A History of Biotechnology (Robert Bud)