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A look at the world today Genetic Engineering (Bilingual Program) Fall, 2010 He, Xiaoyang Dr. Li, Hui Dr. Tian, Shengli Scope Introduction and Molecular Tools Introduction Enzymes vectors Recombinant DNA Technology The study of Gene Expressions Modern Methods in Molecular Biology Schedule 第一章 绪 论 Introduction 2 第二章 基因工程的工具酶 Enzymes 4 第三章 基因工程载体 Vectors 4 第四章 目的基因的获得 2 第五章 基因与载体的体外连接 2 第六章 重组DNA分子导入受体细胞 4 第七章 含目的基因重组体的筛选、鉴定与分析 5 第八章 目的基因的表达 6 第九章 基因工程药物无性繁殖系的组建 3 第十章 基因工程抗体 4 Reading 基因工程 彭银祥等主编 武汉:华中科技大学出版社 ©2006 Gene Cloning and DNA Analysis: An Introduction By Terry A. Brown English Vs. Chinese (PDF) Molecular Biology (Instant Notes, 3rd edition) By Phil Turner, Alexander Mclennan, Andy Bates & Mike White 北京: 科学出版社,2009 References Molecular Cloning A laboratory manual(Third Edition) Sambrook & Russell Cold Spring Harbor Laboratory Press c2001 《分子克隆实验指南》第三版 【作 者】(美)萨姆布鲁克等著 金冬雁等译 【出 版 社】 科学出版社 Molecuolar Biology ( 4th edition ) Robert F. Weaver New York: McGraw Hill Higher Education, c2008 Principles of Gene Manipulation S. B. Primrose, Richard M. Twyman Goals • Groundwork for understanding the experimental and conceptual basis of gene cloning and DNA manipulation • Principles of genetic engineering • Powerful experimental tools and strategies • New discoveries and new methodologies • Perspectives in the literature • Perspectives for the future Evaluation Attendance 10% Assignments 20 ~ 30 % 1. Select a company that sale any biotech products that you like. Then, write a “product review” describing this product in as much detail as possible but no more than one A4 page (or team work presentation?)。 Exam 70 ~ 60 % Close book, 2 hrs. 考试与成绩评定方式 学期总成绩包括出勤、平时成绩和期末考试成绩三部分组成。平时记 录的出勤情况、课堂提问、以及课后作业等占30%,期末成绩占70%。 Chapter 1 Introduction Overview 1.1 1.2 1.3 1.4 Gene and Genetic Engineering A Brief History Applications Safety and Ethical Discussions (伦理学的) 第一章 绪论 教学目的、要求: 1.掌握基因工程的基本概念与研究内容 2.了解基因工程发展简史及与基因工程发展有密切关系的的关键事件 3.了解现代生物技术与基因工程的研究内容和发展趋势 教学内容: 1、基因工程的概念 2、基因工程的发展史 3、基因工程的应用 4、基因工程的安全性与伦理学问题 1.1 Gene and Genetic Engineering 1.1.1 Review Gene: A gene is a basic unit of heredity(遗传) in a living organism. It is "a locatable region of genomic sequence, corresponding to a unit of inheritance, which is associated with regulatory regions, transcribed regions, and or other functional sequence regions “. [1] Coding for proteins 3 classes of genes Coding for RNAs Specific functions Genes hold the information to build and maintain an organism's cells and pass genetic traits to offspring. [2] 1. Pearson H (2006). "Genetics: what is a gene?". Nature 441 (7092): 398–401 2. http://en.wikipedia.org/wiki/Gene Allele(等位基因): Each gene can have different alleles. An allele (from the Greek αλληλος allelos, meaning each other) is one of two or more forms of the DNA sequence of a particular gene. Diploid (二倍体); Triploid (三倍体);etc. The vast majority of living organisms encode their genes in long strands of DNA. The most common form of DNA in a cell is in a double helix structure RNA is common as genetic storage material in viruses, in mammals in particular RNA inheritance has been observed very rarely. Central Dogma of Molecular Biology: The flow of genetic information in the cell starts at DNA, which replicates to form more DNA. Information is then ‘transcribed” into RNA, and then it is “translated” into protein. The proteins do most of the work in the cell. Once information gets into protein, it can't flow back to nucleic acid. Gene Material DNA/RNA Function Nature Peptide coding Reproducible RNA transcripts Movable Regulatory unit Mutagenicity Reconstructible …… 1.1.2 Definition of Gentic Engineering GE: The technology entailing(承担) all processes of altering the genetic material of a cell to make it capable of performing the desired functions, such as producing novel substances. In other words: Genetic engineering is the deliberate(深 思熟虑的), controlled manipulation(操纵,篡 改)of genes in an organism in order to upgrade that organism. 1.2 A Brief History of Genetic Engineering Some major steps in the development of GE Theoretic basis Identification of DNA as the genetic material DNA double helix Central dogma Tools & Tech breakthroughs DNA manipulative enzymes DNA sequencing PCR Plasmid Vector Libraries Bioinformatics Animal Cloning w/t Nuclear Transfer …… Steps marked the beginning of a new age in biology Rediscovery of Mendel's laws helps establish the science of genetics Huntington DNA recombination disease gene mapped to & delivery method chromosome 4 1900 1972 1953 Watson and Crick identify DNA (the double helix) as the Chemical basis of heredity Genetic and physical mapping 1983 1980 DNA markers used to map human disease genes to chromosomal regions 1994-98 1990 Human Genome Projects (HPG) begins-an international effort to map and sequence all the genes in the human genome 1998 Working Draft of the human genome sequencing complete 2000 2005 (or earlier) DNA markers used to map human disease genes to chromosomal regions Gene map expected to be complete Health Policy Research Bulletin, volume 1 issue2, September 2001 1.2.1 Some major steps in the development of GE http://www.nature.com/milestones/miledna/timeline.html 2003 2005 2006 mRNA) Finished the sequence of human genome Finished the sequence of chimpanzee genome Craig C. Mello and Andrew Fire's received a noble prize for RNAi (1998 discovered RNAi degrading 1.2.2 People, Events & Theoretical basis Identification of DNA as the genetic material Factors carryout the genetic messages? locating on the chromatins/chromosomes chromosomes consist of DNA and proteins Which one is the genetic material? Traits ------- Heredity From Mendel to Avery Mendelian Inheritance Parents contribute specific particles (genetic units) to their offspring. ----Implication of gene Genes can exist in several different forms, or alleles. One allele can be dominant over another, so heterozygotes having two different alleles of one Gene will f nerally exhibit the characteristic dictated by the dominant allele. The recessive allele is not lost; it can still exert its influence when pairs with another recessive allele in a homozygote. Johan Friedrich Miescher Swiss Biologist Isolated nuclei of white blood cells in 1869 The major component of “nuclein” is DNA Protein is the other major component of nuclein Led to identification of nucleic acid by Walter Flemming DNA and RNA are both nucleotide polymers Walter Sutton Determined in 1903 that chromosomes carried units of heredity identified by Mendel Wilhelm Johannsen Danish Botanist Named “genes” in 1909 Thomas Hunt Morgan Studied genetics of fruit flies in early 1900’s Experimented with eye color Eye color phenotype was sex-linked His work contributed to the knowledge of X and Y chromosomes Nobel Peace Prize in 1933 for research in gene theory The Chromosome Theory of Inheritance Genes are arranged in linear fashion on chromosome. The reason that certain traits tend to be inheritated together is that the genes governing these traits are on the same chromosome. Every gene has its place (locus) Diploid organism (human) normally have two copies of all chromosomes (except sex chromosomes) DNA recombination occurs in nature Griffith’s Transformation Experiment The discovery of the genetic role of DNA in 1928 two strains of a bacterium, a pathogenic(致病性) “S” and a harmless “R” mixed heat-killed remains of the pathogenic strain with living cells of the harmless strain, some living cells became pathogenic He called this phenomenon transformation, now defined as a change in genotype and phenotype due to assimilation(同化作用) of foreign DNA Living S cells (control) Living R cells (control) Heat-killed S cells (control) Mixture of heat-killed S cells and living R cells RESULTS Mouse dies Mouse healthy Mouse healthy Mouse dies Living S cells are found in blood sample Challenges: “principle “ transform the R into S with smooth coat? Oswald Avery and Colin Macleod In 1944, Oswald Avery, Maclyn McCarty, and Colin MacLeod announced that the transforming substance was DNA Their conclusion was based on experimental evidence that only DNA worked in transforming harmless bacteria into pathogenic bacteria Many biologists remained skeptical, mainly because little was known about DNA Led by the earlier experiment of transfer genetic trait from one train of bacteria to another Avery’s Transformation Experiment Identity the “principle” Animation: http://www.dnaftb.org/dnaftb/17/animation/animation.html Hershey-Chase Bacteriophage Experiment In 1952, Alfred Hershey and Martha Chase performed experiments showing that DNA is the genetic material of a phage known as T2 To determine the source of genetic material in the phage, they designed an experiment showing that only one of the two components of T2 (DNA or protein) enters an E. coli cell during infection 32P is discovered within the bacteria and progeny(子代) phages, whereas 35S is not found within the bacteria but released with phage ghosts(衣壳). They concluded that the injected DNA of the phage provides the genetic information http://glencoe.mcgrawhill.com/sites/9834092339/student_view0/chapter14/hershey_and_chase_experiment .html Phage Radioactive protein Empty protein shell Radioactivity (phage protein) in liquid Bacterial cell Batch 1: Sulfur (35S) DNA Phage DNA Labeled Pr shell Centrifuge Pellet (bacterial cells and contents) Radioactive DNA Batch 2: Phosphorus (32P) Labeled DNA Core Centrifuge Pellet Radioactivity (phage DNA) in pellet Additional Evidence That DNA Is the Genetic Material 1947: Erwin Chargaff- DNA composition varies from one species to the next By 1950s: DNA is a polymer of nucleotides, G=C, A=T Franklin’s X-ray crystallographic images of DNA enabled Watson and Crick to deduce that DNA was helical The X-ray images also enabled Watson and Crick to deduce the width of the helix and the spacing of the nitrogenous bases The width suggested that the DNA molecule was made up of two strands, forming a double helix Double Helix Model of DNA Structure James Watson and Francis Crick Collaborated at Cambridge University and presented the double helix model of DNA structure in 1953 Described DNA dimensions and spacing of base pairs Had major impact on genetic engineering carried out today 1958, 1970 Crick: Central Dogma 1988, Watson: Principle scientist of the HGP Central Dogma Meselson-Stahl DNA must replicate during each cell division 1958 DNA replication: semiconservative model Nirenberg, Ochoa, Khorana 1966 genetic code elucidation Meselson-Stahl Experiments Labeled the nucleotides of old strands with a heavy isotope of nitrogen (15N), new nucleotides were indicated by a lighter isotope (14N). The first replication in the 14N medium produced a band of hybrid (15N-14N) DNA, eliminating the conservative model. A second replication produced both light and hybrid DNA, eliminating the dispersive model and supporting the semiconservative model. http://highered.mcgraw-hill.com/olc/dl/120076/bio22.swf Bacteria cultured in medium containing Bacteria transferred to medium containing 15N 14N Tech supports DNA sample centrifuged after 20 min (after first replication) DNA sample centrifuged after 40 min (after second replication) Less dense Radio labelling More dense First replication Conservative model Semiconservative model Dispersive model Second replication Ultracentrifuge Synchronization 1.2.3 Molecular tools and Technological breakthroughs Enzymes- nucleic acid cleavage, ligation, …… Vector- molecular cloning Polymerase chain reaction DNA sequencing Electrophoretic separation Detection of genes: DNA-Southern blotting; in situ hybridization; FISH technique; RNA- Northern blotting Pr-Western blotting; inmmunohistochemistry Purification Transgenetic organisms …… Discovery of DNA ligase ----the dawn of DNA manipulation DNA recombination happens in the cell — for example, when breaks caused by UV are repaired • • search for an enzyme that could join DNA molecules In this illustration, DNA ligase (in color) encircles the DNAdouble helix. • 1967: The first DNA ligase was purified and characterized in different labs. Enzymatic breakage and joining of deoxyribonucleic acid, I. Repair of single-strand breaks in DNA by an enzyme system from Escherichia coli infected with T4 bacteriophage" , PNAS 57: 10211028. 1967 Making the Cut ----discovery of the restriction enzymes Hamilton Smith and Kent Wilcox • 1970: isolation/characterization of endonuclease R (HindII) from extracts of Haemophilus influenzae (嗜血杆菌)strain Rd. • the enzyme degraded foreign DNA, such as that of phage T7, but did not affect native H. influenzae DNA. • proposed that the enzyme recognizes a specific sequence on the foreign DNA, The 'recombination' potential of restriction enzymes was first demonstrated by Janet Mertz and Ronald Davis. They showed that the R1 restriction endonuclease produces ‘staggered‘(参差) breaks, generating ’cohesive’ ends that are identical and complementary(互补). Their findings suggested that any R1-generated ends can be joined by incubation with DNA ligase to generate hybrid DNA molecules. Thus, the era of recombinant DNA technology was born. key concept : Use of plasmid as vector for shuttling DNA into bacteria 1973, Stanley Cohen and his Stanford colleague Annie Chang, in collaboration with Herbert Boyer and Robert Helling at the University of California in San Francisco, reported the first in vitro construction of a bacterial plasmid. Using EcoR I, they generated fragments from two plasmids (each conferring resistance to one antibiotic), joined them using DNA ligase and applied the mixture to transform E. coli. As they had hoped, a fraction of the transformed bacteria became resistant to both antibiotics while carrying a single hybrid plasmid. Not only had they demonstrated that bacterial plasmids constructed in vitro were functional in bacteria, but they had also described the first plasmid vector. Paul Berg had devised(设计) a similar experiment to transfer foreign DNA into mammalian cells, using the tumour virus SV40 as a vector. In 1972, he made a hybrid molecule in vitro by inserting phage sequences into SV40. These reports immediately raised concerns, as E. coli, which is a natural habitant of the human gut, could now carry hybrid DNA molecules containing SV40 oncogenes(致癌基因) or other potentially harmful sequences. These fears led the community to a self-imposed moratorium(暂停) on recombinant DNA experiments. However, the foundation had been laid and progress soon resumed. Discovery of reverse transcriptase ——Full-length cDNA technologies puzzle: the ability of RNA tumour viruses to stably transform cells without incorporation of a DNA copy of viral genes into the host genome Baltimore, Temin and Mizutani, looked for DNA polymerase activity in purified preparations of such viruses. DNA was being synthesized in RNA-dependent way. Baltimore, D. RNA-dependent DNA polymerase in virions of RNA tumour viruses. Nature 226, 1209–1211 (1970) reverse transcriptase could be used in vitro to synthesize cDNA from mammalian mRNAs. Verma et al. and Kacian et al. both added preparations of glob in mRNAs to reverse transcriptase from avian myeloblastosis virus. They correctly hypothesized that the reaction would only work efficiently if they also added oligo (dT) Reverse transcription has become hugely important in molecular biology. Its influence extends from cloning to the development of microarrays to the annotation of genomes. DNA Libraries: YACs and BACs 1987, 1992 A vector carrying a 50-kb insert was far too small to also contain all regulatory regions. constructing comprehensive libraries covering the whole genomes of higher organisms. Maynard Olson and colleagues exchanged the E. coli plasmid for a yeast artificial chromosome (YAC): a linear DNA molecule that mimics a yeast chromosome, complete with centromere and telomeres. YAC problems: chimaeras of noncontiguous DNA fragments; inserts unstable; purification of YACs proved challenging Challenge: combination of the plasmid simplicity and stability with the aim of adapting it for largefragment cloning. A group led by Melvin Simon modified an endogenous circular plasmid in E. coli, the fertility (F) factor present at one or two copies per cell, to create a cloning vector. In reference to its yeast cousin, they called it bacterial artificial chromosome (BAC). With a cloning capacity of 300 kb, BACs are not as potent as YACs, but they have all the advantages of a bacterial vector: stability, and ease of manipulation and purification. The basic steps in gene cloning . Gene Cloning and DNA Analysis by T.A. Brown. © 2006 T.A. Brown. Cloning allows individual fragments of DNA to be purified. Gene Cloning and DNA Analysis by T.A. Brown. © 2006 T.A. Brown. 1.3 Genetic Engineering Applications Genetic Engineering of Bacteria and Fungi a. Drugs, Vaccines, & Antibiotics b. c. Food Products (e.g., cheese) Metabolism & Biofuel Production d. e. Human Gene Delivery Systems Fermentation & Alcoholic Beverages Genetic Engineering of Animals & Plants a. b. Biopharming (growing transgenic crops to produce pharmaceutical products: drugs, vaccines, proteins) Using Animals to Make Drugs c. Transgenic Crops: corn, soybean, cotton Genetic Engineering of Humans a. b. Germ Cell vs. Somatic Cell Gene Therapy In Vivo Gene Therapy c. d. Ex Vivo Gene Therapy Cloning, Stem Cells, & Gene Therapy The Industry of GE… The most profit… ? 1980 - the U.S. patent for cloning genes is awarded to Cohen and Boyer** First biotech companies formed: 1976 – Genentech (Boyer) 1978 - Biogen 1980 - Amgen 1981 - Immunex 1981 - Chiron 1981 - Genzyme 1.3.1 Agricultural Applications First food made by recombinant DNA technology – Flavr Savr Tomatoes (1994) 60% of foods are genetically modified Solution to world hunger One change in DNA sequence (mutation) can have a significant effect A 601 ACGGTGCCCG CAAAGTGTGG CTAACCCTGA ACCGTGAGGG B 601 ACGGTGCCCG CAAAGTGTGG ATAACCCTGA ACCGTGAGGG + Protein + Herbicide (除草剂) A Herbicide resistant B Herbicide sensitive In the process of causing crown gall disease, the bacterium A. tumefaciens inserts a part of its Ti plasmid — a region called T-DNA — into a chromosome of the host plant. The only vectors routinely used to produce transgenic plants are derived from a soil bacterium called Agrobacterium tumefaciens. This bacterium causes what is known as crown gall disease, in which the infected plant produces uncontrolled growths (tumors, or galls), normally at the base (crown) of the plant. The key to tumor production is a large (200-kb) circular DNA plasmid — the Ti (tumor- inducting) plasmid. When the bacterium infects a plant cell, a part of the Ti plasmid — a region called T-DNA — is transferred and inserted, apparently more or less at random, into the genome of the host plant The functions required for this transfer are outside the T-DNA on the Ti plasmid. The T-DNA itself carries several interesting functions, including the production of the tumor and the synthesis of compounds called opines. Opines are actually synthesized to the host plant under the direction of the T-DNA. The bacterium then uses the opines for its own purposes, calling on opine-utilizing genes on the Ti plasmid. Two important opines are nopaline and octopine; two separate Ti plasmids produce them. Genetically Engineered Crops 3 Types of Resistance • Herbicide Resistance (HR) – Most U.S. crops engineered with resistance to glyphosate • Insect Resistance (IR) – Types of soil bacterium (Bacillus thuringiensis) introduced into plant to target susceptible insects • Virus Resistance The Impact of Genetically Engineered Crops on Farm Sustainability in the United States Division on Earth and Life Study,The National Academes, US April 13, 2010 Genetically Engineering Crops Nationwide acreage of GE soybean, corn, and cotton as a percentage of all acreage of these crops 100 Percent GE crops 80 60 40 All GE Corn Varieties All GE Cotton Varieties 20 All GE Soybean Varieties 0 2000 2001 2002 2003 2004 2005 2006 2007 2008 2009 Year Source: USDA-NASS (2001, 2003, 2005, 2007, 2009b). • Soybeans – Herbicide resistance • Corn – Herbicide resistance – Insect resistance • Cotton – Herbicide resistance – Insect resistance • Oilseed Rape Herbicide resistance Insect resistance Research has mainly focused upon 2 herbicides: glyphosate (Monsanto's Roundup) and glufosinate (Hoechst's Challenge). 1.3.2 Biotechnology and the Environment Waste • Solid: landfills, combustion-including wasteto energy plants, composting • Liquid: septic(腐烂物): sewage treatment • Gas: fossil fuels, chlorofluorocarbons • Hazardous –anything that can explode, catch fire, release toxic fumes, and particles or cause corrosion Biotechnology and the Environment Garbage Test Banana Peel Wood Scrap/Sawdust Wax Paper Styrofoam Cup Tin Can Aluminum Soda Can Plastic Carton Glass Bottles 0.5 Years 4 Years 5 Years 20 Years 100 Years 500 Years 500 Years >500 Years Biogeochemical Cycles are a major part of the recycling process Carbon Cycle: The primary biogeochemical cycle organic cmpds CO2 and back Nitrogen Cycle: proteins amino acids NH3NO2NO3-NO2-N2ON2 NH3 etc_ Sulfur Cycle: Just like the nitrogen cycle, numerous oxidation states. Modeled in the Winogradsky column Phosphorous Cycle: Doesn’t cycle between numerous oxidation states only soluble and insoluble form There is no waste in Nature: From rocks and soil to plants and animals to air and water and back again: Recycled largely by Microbes Bioremediation Basics biodegradation • Aerobic • Oxygen is reduced to water and the organic molecules (e.g. petroleum, sugar) are oxidized • Anaerobic • An inorganic compound is reduced and the organic molecules are oxidized (e.g. nitrate is reduced and sugar is oxidized) • NOTE: Many microbes can do both aerobic and anaerobic respiration; the process which produces the most ATP is used first! Some microorganism of the genera(种属) Nocardia and Pseudomonas can grow in the environment where the hydrocarbons are the only source of food. These bacteria oxidize straight chain aliphatic hydrocarbon such as octane to their corresponding carboxylic acids: E? CH3(CH2)6CH3 +NAD+ O2——– CH3(CH2)6COOH + NADH + H+ China uses oil-eating bacteria to clean up spill Tue Jul 20, 12:09 pm ET BEIJING (AFP) – Authorities in China are using over 23 tones of oil-eating bacteria to help clean up an oil spill in the Yellow Sea caused by a pipeline explosion and fire at the weekend, state media said Tuesday. Petroleum spill in Gulf of Mexico? 1.3.3 Human & Medical Applications Production of Insulin Genetic Screening/Testing Production of experimental mice, oncomouse (cancer mouse) Transgenic animals Stem cells …… Your thinking on biotech companies in Shenzhen? Cloning and Stem Cells Clone created from one cell – not sex cell Cloning – process of creating an identical copy of an original (Example: Identical Twins) Stem cells (master cells) can develop into any tissue Cure their own disease or build new organs – NO REJECTION Stem Cell Differentiation Zygot Adult Five Day Pre-Embryo http://www.nationalgeographic.com/ngm/ in vivo ex-vivo 1.4 Safety and Ethic Issues Facts about GE The result is called genetically modified organism (GMO) One of the major aims is to produce much food at low cost to reduce the world hunger Conglomerates are buying up (收购) biotech start-up companies, seed companies, agribusiness and agrochemical concerns, pharmaceutical, medical and health businesses, and food and drink companies creating life sciences complexes to fashion bio-industrial world Can potentially be used in humans to change their appearance, intelligence, character and adaptability Pros & Cons of GE Is GE precise enough ? Pro arguments: Scientits use „gene guns“ to insert the specific gene in the organism precisely Contra arguments: The choice of gene is precise. But the insertion of this gene into a living cell is imprecise. There is no control where in the DNA the new gene is inserted. This process can disrupt the DNA Pros & Cons of GE Is Genetic Engineering safety? Pro arguments Contra arguments All genetically engineered foods have been thoroughly tested and demonstrated to be safe before they are released into the marketplace Tests are only conducted on animals like rats and mice. Apart from that the scientists are often not independent due to the fact that they are involved into the big companies Genetically engineered foods have been sold in the United States for several years and it is no evidence to indicate that these foods have harmed human health in any way The consequences are now unknown and unanticipated The consequences for the human health can only be assessed after human testing. Pros & Cons of GE Effects on the environment Pro arguments: GE minimizes soil erosion by reducing the need of flowing . Plants resisant to weather , climate insect infestation, desease, molds and fungi. Contra arguments: Every genetically engineered organism released into the environment is a threat to the ecosystem because they are unpredictable by interacting with other living things in the environment, therefore it is difficult to assess the threats of genetically engineered organisms to the ecosystem. GE can create toxins, noxiousvegetation, harm to wild life and may create new molds and fungi. Pros & Cons of GE Effects on the evironment Pro arguments: GE allows the creation of thousands of novel life forms in a brief moment. Contra arguments: Once GMOs are released into the environment they cannot be recalled therefore they are a very dangerous kind of pollution. Pros & Cons of GE Effects on the agriculture Pro arguments: Farmers can spray in order to kill weeds without killing the crops. Contra arguments: Furthermore the weeds might develop their spray resistance and greater herbicide resistance has to be created. The virus-resistance might also create new viruses that never existed before. Pros & Cons in general Genetic engineering reduces costs of production This means that the poor can afford more food Cheaper and safer source of human medicine Higher productivity GE can reduce the World hunger GE is unnatural GE crosses species barriers which would never occur in nature A high danger might be the “gene-flow”transfer of genes from crops to weedy relatives by crosspollination Stem Cell History 1998 - Researchers first extract stem cells from human embryos 1999 - First Successful human transplant of insulin-making cells from cadavers 2001 - President Bush restricts federal funding for embryonic stem-cell research