PRAGATI VIDYA PEETH SCHOOL GWALIOR INVESTIGATORY PROJECT IN BIOLOGY GENETICENGINEERING-A GLANCEATITSLATEST APPLICATIONS PREPARED BY: RAJVEER ATAL CLASS XIICERTIFICATE This is to certify that RAJVEER ATAL of Class XII has satisfactorily completed the project work in Biology prescribed by the Central Board of Secondary Education (CBSE) for AISSSE course for the year 2017-2018. Signature of the teacher: Signature of the invigilator: Name of the Candidate: RAJVEER ATAL Reg. No: Date of Practical Exam:ACKNOWLEDGEMENTS During the course of my project work, I had to encounter quite a few difficulties, which I could overcome thanks to the valuable guidance of my teachers Mrs. Mercy and Mrs.Pravitha, Dept. of Biology, Alfarook Senior Secondary School. | would also like to express my gratitude to my friends for the cooperation and the teamspirit they extended for the successful completion of this project.The century that we left behind has witnessed giant strides in almost all spherescfhuman life. Currently, biotechnology is looked upon as one of the most Promising branches of science, And itis Genetic Engineering that makes most biotechnological applications possible. Genetic engincering is the direct human manipulation ofan organism's genome Using modern DNA technology. It involves the introduction of foreign DNA or synthetic Benes into the organism of interest. The introduction of new DNA does not require the use oflassical genetic methods; however traditional breeding methods are typically uscd for the propagation of recombinant organisms, Humans have altered the genomes of Pecies for thousands of years through artificial selection and more recently mutagene: sis. Genctic engineering as the direct manipulation of PNAby humans outside breeding and mutations has only existed since the 1970s. Humans have altered the genomes of species for thousands of years through artificial selection and more recently mutagenesis, Genetic engineering as the direct manipulation of DNA by humans outside breeding and mutations has only existed since the 1970s, The most Common form of genetic engineering involves the insertion of new genctic material at an Linspecified location in the host genome. This is accomplished by isolating and copying the Genetic material of interest using molecular cloning, methoxls generate a DNA sequence Containing the required genetic elements for expression, and then inserting this construct into the host organism, Other forms of genetic engineering include gene targeting and B outspecific genes via the most common form of gineering involves the insertion of new genctic material at an specified location in the host genome, This is Sfcomplished by isolating and copying the genetic material of interest using mo! hous to Reneratea DNA sequence containing the required get and then inserting this construct into the host organism, engineering alters the genctic makeup ofan organism us reduce heritable material prepared outside the or into a cell thatis then fused or hybridized with nucleic acid (DNA or RNA) techniques to form ‘material followed by the i Vector systey ecular netic elements Genetic ing techniques that ganism either dir eclly into the hostor the host. This involve: sing recombinant Ms of heritable ‘Renetic ial either indirectly through . Macro-injection and micro-Gonetic engineering techniques have been applied in numerous fields including research, agriculture, industry, and medicine. This project seeks to focus on some most promising areas of biotechnological applications... “+ GENETICALLY MODIFIED ORGANISMS Agenetically modified organism (GMO) or genetically enginecred organism (GEO) is organism whose genetic material has been altered using genetic engineering techniques. ‘These techniques, generally known as recombinant DNA technology, use DNA molecules from different sources, which are combined into one molecule to create a new set of genes. This DNA is then transferred into an organism, giving it modified or novel genes. Transgenic organisms, subset of GMOs, are organisms which have inserted DNA that originated in a different species. GMOs are used in biological and medical research, production of pharmaceutical drugs, experimental medicine (c.g, gene therapy),and agriculture (eg. golden rice). The term "genetically modified organism” does not always imply, but can include, targeted insertions of genes from one species into another. For example,a gene from a jellyfish, encoding, a fluorescent protein called GFP, can be physically linked and thus co-expressed with mammalian genes to identify the lacatian of the protein encoded by the GFP-tagged gene in the mammalian cell. Such methods are useful tools for biologists in many areas of research, including those who study the mechanisms of human and other diseases or fundamental biological processes in eukaryotic ar prokaryotic cells. a) Genetically Modified microbes Bacteria were the first organisms to be modified in the laboratory, duc to their simple genetics. These organisms are now used for several purposes, and are particularly important in producing large amounts of pure human proteins for use in medicine. Genetically are used to produce the protein insulin to treat diabetes. Similar bacteria have been used to produce clotting factors to treat hemophilia and human growth hormone to treat various forms of dwarfism. Bacteria synthesize products such as; + Insulin + Hepatitis B vaccine + Tissue plasminogen activator + Human growth hormone+ Icc-minus bacteria + Interferons In materials science,a genetically modified virus has been used to construct more environmentally friendly lithium-ion battery. Gene therapy uses genetically modified viruses to deliver genes that can cure disease into human cells. Although gene therapy is still relatively new, it has had some successes. It has been used to treat genetic disorders such as severe combined immunodeficiency, In 2004, researchers reported thata genetically-moulified virus that exploits the selfish behaviour of cancer cells might offer an alternative way of killing tumours. Genetically modified virus and bacteria b) Genetically Modified Crops ‘Transgenic plants have been engineered to possess several desirable traits, such as resistance to pests, herbicides, or harsh environmental conditions, improved product shelf life, and increased nutritional value. Since the first commercial cultivation of genetically modified plants in 1996, they have been modified to be tolerant to the herbicides glufosinate and glyphosate, to be resistant to virus damage as in Ring spot virus- resistant GM papaya, grown in Hawaii, and to produce the [it toxin, an insecticide that is non-toxic to mammals. Most GM crops grown today have been modified with "input traits", which provide benefits mainly to farmers. Golden Riceis a transgenic variety of rice, with genes for the synthesis of b-carotene taken from the temperate garden favourite Narcissus pseudonarcissus (daffodil) and inserted into the genome of a temperate strain of rice, using Agrobacterium tumefaciens as the vector, to effect the transfer. ‘The gene construct also contains some genes for enzymes af the biosynthetic pathway of b-carotene, from another bacterium Erwinia uredovora,benctic engineering techniqu more precise than mutagenesis (mutation breeding) whe radiation or chemicals to create a non-specific but stable Which humans modify food organisms include selective animal breeding, andsomaclonal variation. GM foods were first put on the market in 1996, Typically, are transgenic plant products: soybean, corn, canola, and have also been developed, although as of July 2010 none 2006 pig was controversially engineered to Produce omega-3 fatty acids through the expression of a roundworm gene, Researchers have also developed a genetically-modified breed of pigs that are able to absorb plant phosphorus more efficiently, andasa Consequence the phosphorus content oftheir manure is reduced by as much as GO%t4. rites have abjected to GM foods on several grounds, including safety 'ssues ecological concerns, and economic concerns ralsed by the fact these organisms subject to intellectual property law, These techniques are much Te an organism is exposed to change. Other techniques by breeding; plant breeding, and Renetically modified foods cotton seed oil. Animal products are currently on the market2l In are Genetically Engineered Roses¢) Transgenic Animals Atransgenic animal is one that carries a foreign gene that has been deliberately inserted Into its genome. The foreign gene is constructed using recombinant DNA methodology. In addition to the gene itself, the DNA usually includes other sequences to enable it to be Incorporated into the DNA of the host and to be expressed correctly by the cells of the host. Transgenic animals are used as experimental models to perform phenotypic and for testing in biomedical research. Genetically Modified (Genetically Engineered) animals are becoming more vital to the discovery and development of cures and treatments for many serious diseases. By altering the DNA or transferring DNAto an animal, we can develop certain proteins that may be used in medical treatment. Stable expressions of human proteins have been developed in many animals, including sheep, pigs, and rats. Some examples are: Human-alpha-1-antitrypsin2! which has been developed in sheep and is used in treating humans with this deficency and transgenic pigs with human-histo- compatibility have been studied in the hopes that the organs will be suitable for transplant with less chances of rejection. Transgenic livestock have been used as bioreactors since the 1990s. Many medicines, including insulin and many imnumizations are developed in transgenicanimals.| In March 2011, the bioactive recombinant Human Lysozyme was expressed in the milk of cloned transgenic cattle, This field is growing rapidly and new pharming uses are being discovered and developed. The extent that trangenic animals will be useful in the medical field as well as other fields is very promising based on results thus far. The Glow Fish,a fluorescent red zebra fish sold as a novel pet, has become the first al sold to US. consumers. In 1999, Dr. Zhiyuan Gong and his colleagues at transgenic anim: i called green fluorescent the National University of Singapore were working with a gene i protein (GFP), originally extracted froma jellyfish, that naturally produced bright green fluorescence. They inserted the gene into zebra fish embryo, allowing it to integrate into the zebrafish's genome, which caused the fish to be brightly fluorescent under both hite light and ultraviolet light. Their goal was to developa fish that could natural wtdetect pollution by selectively fluorescing in the presence of environmental toxins. The developmentof the constantly fluorescing fish was the first step in this process. Shortly thereafter, his team developed a line of red fluorescent zebra fish by adding a gene froma sea coral,and orange-yellow fluorescent zebra fish, by adding a variant of the jellyfish gene, Later, a team of researchers at the National University of Taiwan, headed by Professor Muai-Jen Tsai succeeded in creating a medaka (rice fish) with a fluorescent green color, Which like the zebrafish is a model organism used in biology, Transgenic: Rat Fruit flies \n biological research, transgenic fruit Mies (Drosophila melanogaster) are model organisms used to study the effects of genetic changes on development [Fruit ies areoften Preferred aver other animals due to their s hort life cycle, low maintenance requirements, and relatively simple genome compared to many vertebrates, Mosquitoes 'n 2010, scientists created *malaria-resistant Mosquitoes” in the laboratory. The World Health Organization estimated that Malaria killed almost one million people in 2008. Genetically modified male mosquitoes conta in order to combat the spread of Dengue important carrier of dengue fever, were ‘mosquitoes in the Cayman Islands. Arou fever every year and 40,000 people die Mammals Genetically modified mammals are an important category of genetically modified Creinisms. Transgenic mice are often used to study cellular and tissue to disease. ining a lethal gene have been developed fever. Aedes aegypti mosquitoes, the single most reduced by 80% in a 2010 trial of these GM nd 50- 100 million people are affected by Dengue fromit. -specifle responsesIn 1999, scientists at the University of Guelph in Ontario, Canada created the genetically engineered Enviropig. The Enviropig excretes from 30 to 70.7% less phosphorus in manure depending upon the age and dict.In February 2010, Environment Canada determined that Enviropigs are in compliance with the Canadian Environmental Protection Act and can be Produced outside of the research contextin controlled facilities where they are segregated from other animals. In 2009, scientists in Japan announced that they had successfully transferred a gene into a primate species (marmosets) and produced a stable line of breeding transgenic primates for the first time, Their first research target for these marmosets was Parkinson's disease, but they were also considering Amyotrophic lateral sclerosis and Huntington's disease. In 2011,scientists in China released news that they have introduced human genes into 300 dairy cows to produce milk with the same properties as human breast milk. Aside from milk production, the researchers claim these transgenic cows to be identical to regular cows! Cnidarians Cnidarians such as Hydra and the sea anemone Nematostella vectensis have become attractive model organisms to study the evolution of immunity and certain developmental processes. An important technical breakthrough was the development of procedures for generation of stably transgenic hydras and sea anemones by embryo microinjection, Hybrid Animals “* GENETIC ENGINEERING IN MEDICINE Genetic engineering is becominga major force in conventional medicine. It has got numerous applications in medicine ranging from vaccines to transgenic organ transplants. Na) The Artificial Blood ‘The artificial blood is a genetically engineered form of hemoglobin, the complicated protein that enclosed in red blood cells~carries oxygen from the lungs to tissues throughout the body. Many companies have been searching for an artificial blood because of the annual worldwide shortage of about 100 million units of blood and the military's need for blood replacements that can be stored in field conditions without refrigeration. An artificial blood also would virtually eliminate the risk of contracting AIDS, hepatitis and other viral diseases through transfusions. Development of emulsion technologies resulted in the production of compounds which utilized smaller chain perfluoracarbon molecules to more effectively emulsify the perfluorocarbons, allowing higher concentrations of active agent in the emulsion and thus higher oxygen carrying capabilities. The Improved stability of the newer emulsions are vastly superior to the first generations of perfluorocarhons; current emulsions can be stored at 4°C for extended periods of time (months) without appreciable degradation of activity. Artificial Haemoglobin b) Cloned Pigs Modified for Use in Human Transplants Two competing teams have cloned pigs that have been genetically modified to produce organs mare suitable for transplantation into humans, re Organs are well suited for transplantation; they are approximately the same size as human organ nd have Sinuilar plumbing, which makes reconnecting blood vessels much cnr ti cof ters tends to be large and pigs reproduce quickly, raising the grass eres nply of “spare” organs. A problem with using pig organs, however, is it ated with sugar molecules that trigger acute rejection in people. H antibodies attach themselves to these sugar molecules and quickly destroy the needy transplanted pig organ. To circumvent the rejecti ists are working to produce pigs ie i ejection, scientists are working to produce pig q¢) Genetically Engineered FSH FSIlis produced by the pituitary gland and directly stimulates the ovaries to recruit and SUPNOr ovarian follicles, each containing ane egg. The hypothalamus adjusts the Production of FSH depending upon the levels af other hormones such ag estrogen. FSH is used in stimulated 101 and assisted reproductive technology cycles (IVF) because it Ruuses the development of numerous follicles. More follcies are needed in ART cycles because some do not fertilize or do not continue to develop. There are several FSH Products available in the United States including Follistim, Gonal-F, Bravelle, and Repronex, Follstim and Gonal- are produced by genctic engineering using manmotion culture cells, ‘These products are identical to the body's FSH. Bravelle and Repronex are derived from the urine of postmenopausal women. BIOFUEL- AN ALTERNATIVE Algae fuel might be an alternative to fossil fuel and uses algaeas its source of natural eposits, Several companies and government agencies are funding efforts to reduce capital and operating casts and make algae fuel production commercially viable Harvested algae, Uke fossil fuel, release COz when burnt but unlike fossilfucl the COs Is taken outof the auinosphere by the growing algae. High oi prices, competing demands between fonds and other biofuel sources, and the world fond crisis, have ignited interest in alga culture(farming algae) for making biodiesel, biocthanol, biagasoline, biomethanol, blobutanoland other biofuels, using land that is not suitable for agriculture Biodiesel Cure ly most research into efficient algal-oil production is being done in the private Sector, but predictions from small scale production expe iments bear out that using algae to produce biodiese! may be the only viable method by which to produce enough automotive fuelto replace current world diesel usage. I algae-derived biodiesel were to Feplace the annual global production of 1.1bn tons of conventional diesel, a land mass of 57.3 million hectares would be required, This compares highly favourable to other biofuels Microalgae have much faster growth rates than terrestrial ero ps.The per unit area yield of oilfrom algae is estimated to be from between 5,000 to 20,000 US gallons per pace Per year (4.700 to 18,000 m?/km?-a).U21 This is 7 to 30 times greater than the next best crop, Chinese tallow (700 US gal/acre-a oF 650 m1/km2a), Studies show that some species of algae can produce up to 60% of their dry weight in the form of ol. Because the cells graw in aqueous suspension, where they have more efficient secass to Water, COz and dissolved nutrients, microalgae are eapable of produicing large amounts of biomass and usable oil in either high rate algal ponds or photo bioreactors, Thisoil can then be turned into biodiesel which could be sold for use in automobiles. Regional production of microalgae and processing into biofuels will provide economic benefits to rural communities. LL The National Algae Biofuel Technology Roadmap_emphasizes the role of genetic engineering to find, adjust and deploy optimized algae species for biofuel development. » BIOWEAPON - A CHALLENGE Rapid developments in biotechnology, genetics and genomics are undoubtedly creating a variety of environmental, ethical, political and social challenges for advanced societies. But they also have severe implications for international peace and security because they open up tremendous avenues for the creation of new biological weapons. The genetically engineered 'superbug'—highly lethal and resistant to environmental influence or any medical treatment—is only a small part of this story. Much more alarming, from an arms~ control perspective, are the possibilities of developing completely novel weapons on the basis of knowledge provided by biomedical research—developments that are already taking place. Such weapons, designed for new types of conflicts and warfare scenarios, secret operations or sabotage activities, are not mere science fiction, but are increasingly becoming.a reality that we have to fac The history of biological warfare is nearly as old as the history of warfare itself. In ancient times, warring parties poisoned wells or used arrowheads with natural toxins. By using genetic engineering, biological researchers have already developed new weapons thatare much more effective than their natural counterparts. Countless examples from the daily work of molecular biologists could be presented here, not least the introduction of, antibiotic resistance into bacterial pathogens, which today is routine work in almost any microbiology laboratory. Indeed, many research projects in basic science show— sometimes unwillingly and unwittingly —how to overcome current scientific and. technological limits in the military use of pathogenicagents. Furthermore, genctic engincering is not merely a theoretical possibility for future biowarfare:it has already beenapplied in past weapons programmes, particu is the UssR's ‘invisible anthrax’, resulting from. the introduction of, {nto Bacillus anthracis that altered its immunological propertie. Existing vaccines proved to be ineffective Against this new ‘Rent larly in the former Soviet Union, One example analien gene The global norm ag, the 1972 Biological countries have be This mor, ainst biological weapons and Toxin Weapons Cony: en en} al barrier se aid down in the 1925 Geneva Conve: tion, cle: ntionand ‘arly contributed to the fact that few Baged in research into offensive biowartare during recent decades, Shs tobe lower for ‘non-lethat weapons thet are targeted against materials or drug-producing plants, Indeed. today’s technical possibilities Rew interest in this area that might he leading to anew biological New technological possibilities millitary concepts in the interest in weapons that, until recently, h {mn 1998, it became public that the US Naval Research Laboratory in Wash developing genetically engineered fungi with offensive biowarfare Potential. They isolated natural microorganisms that degrade + variety of ‘metals, and use, materials, such as plastics, rubber and ‘dgenctic engineering to make them More powerful and focus these genetically engineered microtise ‘can dest troy military paints in 72 hour: Microbes that are comy are creating a arms race USA and led toa renewed and rejected, ington DC was ed —one of s. as biological warfare ag Agents,CONCLUSION We have discussed the promising aspects of Genetic Engincering that can bring about tremendous changes in human life. However the manipulation of living organisms by the human race cannot go on any further without regulation, Some ethical standards are required to evaluate the morality of all human activities that might help or harm living organisms. Going beyond the morality of such issucs the biological significance of such things is also important. Genetic modification of organisms can have unpredictable results when such organismsare introduced into the ecosystem. Every new technology aims to Improve man’slife. It is for man to make the judicious use of its applications...BIBLIOGRApHY > : > ei > Lancet magazine(July2011 issue)