Genetic Engineering

Genetic engineering, also called genetic modification, is the direct manipulation of an organism's genome using biotechnology. New DNA may be inserted in the host genome by first isolating and copying the genetic material of interest using molecular cloning methods to generate a DNA sequence, or by synthesizing the DNA, and then inserting this construct into the host organism. Genes may be removed, or "knocked out", using a nuclease. Gene targeting is a different technique that uses homologous recombination to change an endogenous gene, and can be used to delete a gene, remove exons, add a gene, or introduce point mutations.

Applications of Genetic Engineering

The use of recombinant DNA technology has become common as new products from genetically altered plants, animals, and microbes have become available for human use. In 1997, Dolly made headlines as the first successfully cloned large mammal (sheep). Since then there have been many similar advances in medicine, such as treatments for cancer; many advances in agriculture, such as transgenic insect-resistant crops; and many advances in animal husbandry, such as growth hormones and transgenic animals (an animal that has received recombinant DNA). An organism that is generated through genetic engineering is considered to be a genetically modified organism (GMO). The first GMOs were bacteria in 1973; GM mice were generated in 1974. Insulin-producing bacteria were commercialized in 1982 and genetically modified food has been sold since 1994. Glofish, the first GMO designed as a pet, was first sold in the United States December in 2003. Most biotechnologists envision DNA technological applications as one of the new frontiers in science with tremendous growth and discovery potential.
Genetic engineering has applications in medicine, research, industry and agriculture and can be used on a wide range of plants, animals and micro organisms

Medicine
Some of the most promising and powerful applications of genetic engineering are in the field of medicine. Researchers are using it to diagnose and predict disease, and to develop therapies and drugs to treat devastating diseases like cancer, Alzheimer's, diabetes, and cystic fibrosis. Explore more ways genetic engineering techniques can be used for medical purposes. Genetic engineering has resulted in a series of medical products. The first two commercially prepared products from recombinant DNA technology were insulin and human growth hormone, both of which were cultured in the E. coli bacteria. Since then a variety of products have appeared in the market, including the following abbreviated list, all made in E. coli:

Genetically Engineered Pharmaceuticals

Tumor necrosis factor. Treatment for certain tumor cells Interleukin-2 (IL-2). Cancer treatment, immune deficiency, and HIV infection treatment Prourokinase. Treatment for heart attacks Taxol. Treatment for ovarian cancer Interferon. Treatment for cancer and viral infections

factor VIII for males suffering from hemophilia A factor IX for hemophilia B human growth hormone (GH) erythropoietin (EPO) for treating anemia three types of interferons - fight viral infections several interleukins granulocyte-macrophage colony-stimulating factor (GM-CSF) for stimulating the bone marrow after a bone marrow transplant tissue plasminogen activator (TPA) for dissolving blood clots adenosine deaminase (ADA) for treating some forms of severe combined immunodeficiency (SCID) angiostatin and endostatin for trials as anti-cancer drugs

parathyroid hormone
Medicinal Applications of Genetic Engineering : Medicinal Applications of Genetic Engineering Human insulin genes have been implanted in bacteria which, therefore, become capable of synthesizing insulin. Bacterial insulin is identical to human insulin, since it is coded by human genes. Diabetics have been receiving bacterial insulin in test programmes, and it appears to be as effective as insulin from animal sources.***Growth hormone Hormones

Production of Human Insulin- : Production of Human Insulin-

Vaccines : Vaccines Gardisil or Silgard is a vaccine proven to prevent certain types of human papillomavirus (HPV) specifically HPV types 16, 18, 6, and 11 ***Vaccines against malaria and hepatitis may also be produced in the near future.

Interferons- : Interferons- Interferons are virus induced proteins having anti-viral in action. ****Interferon provides natural defence against such viral diseases as hepatitis and influenza. It also appears to be effective against certain types of cancer, especially cancer of the breast and lymph nodes.

:: : Alpha () thalassemias :Deletion of the 16p chromosome. thalassemias result in decreased alpha-globin production ***Beta thalassemias are due to mutations in the HBB gene on chromosome 11 . Hereditary diseases:Thalassemias

Sickle cell disease : Sickle cell disease The specific molecular defect was identified as a single amino acid substitution of valine for glutamic acid at position 6 of the beta-globin polypeptide chain. genetic mutation in the globin gene as a change in the codon GAG to GTG.

Treatment : Treatment Non-defective genes could be transferred into the bone marrow along with the methotrexate-resistance genes. Treatment with methotrexate would then destroy the cells containing defective genes and would permit the non-defective cells to form bone marrow

Haemophilia:- : Haemophilia:- Factor replacement can be either isolated from human blood serum, recombinant, or a combination of the two

Cystic Fibrosis - : Cystic Fibrosis - There are several mechanisms mutations caused problems with the cystic fibrosis transmembrane conductance regulator [CFTR ]protein. F508, for instance, creates a protein that does not fold population result in proteins that are too short because production is ended prematurely. that do not use energy normally, do not allow chloride, iodide and thiocyanate to cross the membrane appropriately, or are degraded at a faster rate than normal.

Treatment : Treatment *****Gene therapy to treat cystic fibrosis involves the sequencing and cloning of the CFTR gene

Huntingtons Chorea : Huntingtons Chorea trinucleotide repeat disorders cytosine-adenine-guanine (CAG)repeated multiple times (i.e. ... CAGCAGCAG ...), production of series of chain of glutamine known as a polyglutamine tract (or polyQ tract), and the repeated part of the gene, the PolyQ region.[14]

Huntingtons Choria : Huntingtons Choria Huntingtons Chorea TREATMENT

Other Medical Applications:- : Other Medical Applications:- Enzymes. The enzyme urokinase, which is used to dissolve blood clots, has been produced by genetically engineered microorganisms Antibodies. One of the aims of genetic engineering is the production of hybridomas. These are long lived cells that can produce antibodies for use against disease

Urofollitropin is a purified form of follicle-stimulating hormone (FSH) that is manufactured by extraction [1] from human urine and then purified to remove various proteins and other substances. FSH is important in the development of follicles (eggs) produced by theovaries. Given by subcutaneous injection, it is used [2] in combination with human chorionic gonadotropin (hCG) to assist in ovulation andfertility. It is also used [2] with in vitro fertilization methods. The dosage is adjusted to each individual's response. The most common side effects are abdominal or pelvic pain, bloating, as well as redness, pain or swelling at the injection site. Follitropin is possibly associated with increased risk of endometrial carcinoma. It is not for use during pregnancy, as there is evidence for birth defects under follitropin treatment

Medicine[edit]
In medicine genetic engineering has been used to mass-produce insulin, human growth hormones, follistim (for treating infertility),human albumin, monoclonal antibodies, antihemophilic [69][70] factors, vaccines and many other drugs. Vaccination generally involves injecting weak live, killed or [71] inactivated forms of viruses or their toxins into the person being immunized. Genetically engineered viruses are being developed that can still confer immunity, but lack [72] the infectious sequences. Mouse hybridomas, cells fused together to create monoclonal antibodies, [73] have been humanised through genetic engineering to create human monoclonal antibodies. Genetic [74][75] engineering has shown promise for treating certain forms of cancer. Genetic engineering is used to create animal models of human diseases. Genetically modified mice are [76] the most common genetically engineered animal model. They have been used to study and model cancer (the oncomouse), obesity, heart disease, diabetes, arthritis, substance abuse, anxiety, aging and [77] Parkinson disease. Potential cures can be tested against these mouse models. Also genetically modified pigs have been bred with the aim of increasing the success of pig to human organ [78] transplantation. Gene therapy is the genetic engineering of humans by replacing defective human genes with functional copies. This can occur insomatic tissue or germline tissue. If the gene is inserted into the germline tissue [79][80] it can be passed down to that person's descendants. Gene therapy has been successfully used to [81] treat multiple diseases, including X-linked SCID, chronic lymphocytic [82] [83] leukemia(CLL), and Parkinson's disease. In 2012, Glybera became the first gene therapy treatment to be approved for clinical use in either Europe or the United States after its endorsement by the [84][85] European Commission. There are also ethical concerns should the technology be used not just for treatment, but for enhancement, modification or alteration of a human beings' appearance, adaptability, [86] intelligence, character or behavior. The distinction between cure and enhancement can also be difficult [87] to establish. Transhumanists consider the enhancement of humans desirable.

History of genetic engineering

Artificial selection: Humans have been doing this for centuries, breeding certain plants and animals for their desirable traits. Horse breeding is evidenced as far back at 2500 BC. 1972 First bacteria is genetically engineered combining DNA from the monkey virus SV40 with the lambda virus. 1973 - First transgenic organism created by adding antibiotic resistant genes to E. coli bacterium. 1974 First transgenic animal created by introducing foreign DNA into a mouse. 1976 First company created: Genetech 1980 Supreme Court ruled that created life can be patented.

HUMAN GROWTHHORMONE INTRODUCTION :-In the human body themalfunctioning of growth hormonethat is SOMATOSTATIN andSOMATOTROPHIN leading to painfuland disabling disorders such asAcromegaly(uncontrolled bonegrowth), and Dwarfism. SYNTHESIS OF GROWTHHORMONE SOMATOSTATIN:- It was the firsthuman protein synthesized in E.coli . It is very short protein,only 14 aminoacid in length thats why it wassuited for artificial gene synthesis. SOMATOTROPHIN:- This protein is191 amino acid long. cDNA cloning was used to obtain asomatotrophin-producing E.coli stain.

Agriculture Crop plants have been and continue to be the focus of biotechnology as efforts are made to improve yield and profitability by improving crop resistance to insects and certain herbicides and delaying ripening (for better transport and spoilage resistance). The creation of a transgenic plant, one that has received genes from another organism, proved more difficult than animals. Unlike animals, finding a vector for plants proved to be difficult until the isolation of the Ti plasmid, harvested from a tumor-inducing (Ti) bacteria found in the soil. The plasmid is shot into a cell, where the plasmid readily attaches to the plant's DNA. Although successful in fruits and vegetables, the Ti plasmid has generated limited success in grain crops. Creating a crop that is resistant to a specific herbicide proved to be a success because the herbicide eliminated weed competition from the crop plant. Researchers discovered herbicide-resistant bacteria, isolated the genes responsible for the condit ion, and shot them into a crop plant, which then proved to be resistant to that herbicide. Similarly, insect-resistant plants are becoming available as researchers discover bacterial enzymes that destroy or immobilize unwanted herbivores, and others that increase nitrogen fixation in the soil for use by plants. Geneticists are on the threshold of a major agricultural breakthrough. All plants need nitrogen to grow. In fact, nitrogen is one of the three most important nutrients a plant requires. Although the atmosphere is approximately 78 percent nitrogen, it is in a form that is unusable to plants. However, a naturally occurring rhizobium bacterium is found in the soil and converts atmospheric nitrogen into a form usable by plants. These nitrogen-fixing bacteria are also found naturally occurring in the legumes of certain plants such as soybeans and peanuts. Because they contain these unusual bacteria, they can grow in nitrogen-deficient soil that prohibits the growth of other crop plants. Researchers hope that by isolating these bacteria, they can identify the DNA segment that codes for nitrogen fixation, remove the segment, and insert it into the DNA of a profitable cash crop! In so doing, the new transgenic crop plants could live in new fringe territories, which are areas normally not suitable for their growth, and grow in current locations without the addition of costly fertilizers! Animal Husbandry Neither the use of animal vaccines nor adding bovine growth hormones to cows to dramatically increase milk production can match the real excitement in animal husbandry: transgenic animals and clones. Transgenic animals model advancements in DNA technology in their development. The mechanism for creating one can be described in three steps: 1. Healthy egg cells are removed from a female of the host animal and fertilized in the laboratory. 2. The desired gene from another species is identified, isolated, and cloned. 3. The cloned genes are injected directly into the eggs, which are then surgically implanted in the host female, where the embryo undergoes a normal development process. It is hoped that this process will provide a cheap and rapid means of generating desired enzymes, other proteins, and increased production of meat, wool, and other animal products through common, natural functions. Ever since 1997 when Dolly was cloned, research and experimentation to clone useful livestock has continued unceasingly. The attractiveness of cloning is the knowledge that the offspring will be genetically identical to the parent as in asexual reproduction. Four steps describe the general process: 1. A differentiated cell, one that has become specialized during development, with its diploid nucleus is removed from an animal to provide the DNA source for the clone.

2. An egg cell from a similar animal is recovered and the nucleus is removed, leaving only the cytoplasm and cytoplasm organelles. 3. The two egg cells are fused with an electric current to form a single diploid cell, which then begins normal cell division. 4. The developing embryo is placed in a surrogate mother, who then undergoes a normal pregnancy.

Agriculture Crop plants have been and continue to be the focus of biotechnology as efforts are made to improve yield and profitability by improving crop resistance to insects and certain herbicides and delaying ripening (for better transport and spoilage resistance). The creation of a transgenic plant, one that has received genes from another organism, proved more difficult than animals. Unlike animals, finding a vector for plants proved to be difficult until the isolation of the Ti plasmid, harvested from a tumor-inducing (Ti) bacteria found in the soil. The plasmid is shot into a cell, where the plasmid readily attaches to the plant's DNA. Although successful in fruits and vegetables, the Ti plasmid has generated limited success in grain crops. Creating a crop that is resistant to a specific herbicide proved to be a success because the herbicide eliminated weed competition from the crop plant. Researchers discovered herbicide-resistant bacteria, isolated the genes responsible for the condition, and shot them into a crop plant, which then proved to be resistant to that herbicide. Similarly, insect-resistant plants are becoming available as researchers discover bacterial enzymes that destroy or immobilize unwanted herbivores, and others that increase nitrogen fixation in the soil for use by plants. Geneticists are on the threshold of a major agricultural breakthrough. All plants need nitrogen to grow. In fact, nitrogen is one of the three most important nutrients a plant requires. Although the atmosphere is approximately 78 percent nitrogen, it is in a form that is unusable to plants. However, a naturally occurring rhizobium bacterium is found in the soil and converts atmospheric nitrogen into a form usable by plants. These nitrogen-fixing bacteria are also found naturally occurring in the legumes of certain plants such as soybeans and peanuts. Because they contain these unusual bacteria, they can grow in nitrogen-deficient soil that prohibits the growth of other crop plants. Researchers hope that by isolating these bacteria, they can identify the DNA segment that codes for nitrogen fixation, remove the segment, and insert it into the DNA of a profitable cash crop! In so doing, the new transgenic crop plants could live in new fringe territories, which are areas normally not suitable for their growth, and grow in current locations without the addition of costly fertilizers!

Agriculture : Agriculture -food processors affected by genetic engineering. -shelf-life, storage, food-handling;extended and simplified. -help resist spoilage. plants transformed-insect,disease, and herbicide resistant.-animals treated engineered hormones-produce more milk, leaner meat.

Animal Husbandry Neither the use of animal vaccines nor adding bovine growth hormones to cows to dramatically increase milk production can match the real excitement in animal husbandry: transgenic animals and clones. Transgenic animals model advancements in DNA technology in their development. The mechanism for creating one can be described in three steps: 1. Healthy egg cells are removed from a female of the host animal and fertilized in the laboratory. 2. The desired gene from another species is identified, isolated, and cloned. 3. The cloned genes are injected directly into the eggs, which are then surgically implanted in the host female, where the embryo undergoes a normal development process.

It is hoped that this process will provide a cheap and rapid means of generating desired enzymes, other proteins, and increased production of meat, wool, and other animal products through common, natural functions. Ever since 1997 when Dolly was cloned, research and experimentation to clone useful livestock has continued unceasingly. The attractiveness of cloning is the knowledge that the offspring will be genetically identical to the parent as in asexual reproduction. Four steps describe the general process: 1. A differentiated cell, one that has become specialized during development, with its diploid nucleus is removed from an animal to provide the DNA source for the clone. 2. An egg cell from a similar animal is recovered and the nucleus is removed, leaving only the cytoplasm and cytoplasm organelles. 3. The two egg cells are fused with an electric current to form a single diploid cell, which then begins normal cell division. 4. The developing embryo is placed in a surrogate mother, who then undergoes a normal pregnancy.

5.

Applications of Genetic Engineering in Biotechnology

6. The basic principle of genetic engineering is gene transfer, achieved by various methods to produce recombinant proteins, genetically modified microorganisms, transgenic plants and transgenic animals for commercial application. Genetic engineering, thus ultimately influences the growth of biotech industry. The two significant feature of genetic engineering is production of beneficial proteins and enzymes in surplus quantities and creation of transgenic plants, transgenic animals and genetically modified microorganisms with new characters beneficial for themselves using recombinant DNA technology. The discovery of a new protein either with a therapeutic property or application in food industry by a researcher or scientist would not have reached humans, for the use by humans without the application of genetic engineering in mass producing such proteins. Recombinant proteins production and uses: The industrial production of proteins is done by transferring the desired gene responsible for the particular protein to be manufactured from the source organism to the preferred host organism through recombinant DNA technology. The host organism can be a bacteria or a eukaryote. The most preferred bacterial host is Escherichia coli for industrial production of proteins. The well established gene structure, faster growth rate, easy to cultivate and handle are the salient features of the E. coli bacterium fascinated the bio technologists to use this in recombinant protein production. Besides all these commendable characters of E. coli, the final output product is found to be unstable and difficult to purify. As a result research encouraged the use of eukaryotic host like yeast, cells of insects and cells of mammals in protein production. The proteins produced in this way find its way into pharmaceutical industry and food industry. The recombinant proteins produced in the industry using the techniques of genetic engineering acts as drugs for various human diseases. To name a few, insulin produced for diabetes, alpha 1- antitrypsin in treating emphysema, calcitonin to treat rickets, interferon to treat viral infections and cancer, Factor VIII for hemophilia, production of growth hormone to act against growth retardation and chorionic gonadotrophin in the treatment of infertility. Some of the industrial manufactured enzymes occupy a vital position in the food industry. For example, the recombinant

enzymes like rennin and lipase are used in cheese making, the role of alphaamylase in beer industry, the antioxidant property of the industrially produced enzyme catalase and the use of protease in detergents. Uses of Transgenic plants: In order to improve the quality and quantity of plants, traditional method of plant breeding is replaced by the creation of transgenic plants. The transgenic plants are plants carrying foreign genes introduced deliberately into them to develop a new character useful for the plant. The infection of plants by microorganism mostly viruses, poor production and decline in quality of plants due to attack by insects and the plants inability to withstand the pesticide or the weedicide used in the agriculture process welcomed the genetic engineering technology to develop transgenic plants with new characters like resistance to infections, defensive against the attacking insects and resistance to pesticides or weedicide. The transfer of gene responsible for the protein protoxin from Bacillus thuringiensis to plants to develop resistance against the attacking insects is a remarkable example. Also the digestive action of the insects on the plants is restricted or inhibited by transfer of gene responsible for a particular protein with the property to arrest protease activity. The pesticides and weedicides used to destroy the pests and weeds is also a threat to the cultivated plants. The effects of such chemicals are alleviated by developing a new character called resistance to chemicals in plants. Development of resistance in plants against the weedicide glyphosate states the role of genetic engineering in plant breeding. Uses Transgenic animals: Transgenic animals are animals carrying foreign genes deliberately introduced into them and exhibiting the characteristics of the introduced gene. Animals are suitable for various research activities trying to help mankind. In that way transgenic animals are created to study human diseases to derive appropriate treatment methods and to develop and identify the drug useful to treat the disease. The presence of human proteins in milk of animals is made possible by genetic engineering. Gene transfer is done in animals to increase the milk production and to increase the growth. Like a coin has two sides, the other face of genetic engineering like creation of genetically modified organisms to be used as biological weapons is not welcoming.