Biotechnology

Describe how biotechnology dates back to 6000 BCE.

- Neolithic cultures fermented grapes to make wine. - Babylonians used microbial yeast for bread production.

Discuss new methods of pest management.

- New methods of pest management use genetic engineering of crops to kill of prevent insects from even being a pest. This involves the development of new crop genotypes toward safe and sustainable agriculture. - This is a chemically free approach and is also economically viable.

Define biotechnology

Biotechnology can be defined as any application that uses biological systems, living organisms or derivatives thereof, to make or modify products or processes for specific use by humans.

How are GMO blood clotting factors made.

Factor IX preparation - plasmids containing the Factor IX gene and plasmids with a gene that confer methotrexate resistance were transferred into Chinese hamster ovary cells. - in order to obtain a stable transfection, cells that do not integrate the transfected DNA are destroyed. - Transfected cells are grown in increasing concentrations of methotrexate, promoting their survival and stability while diminishing the survival of other cells. Factor VIII preparation - plasmid-derived factor VIII was heat treated, lyophilized factor VIII concentrate prepared from a donor pool screened for antibodies to HIV. - recombinant (r} factor VIII was a lyophilized product prepared in Chinese hamster ovary cells transfected with DNA containing the coding sequence for human factor VIII fused to a strong mammalian promoter. - factor VIII was purified to homogeneity by affinity chromatography with use of a monoclonal antibody to human factor VIII and ion-exchange chromatography.

Discuss the concept of spider silk farms.

- The dragline form of spider silk is regarded as the strongest material known, being 5 times stronger than steel. Material can be used to make lighter, stronger bulletproof vests, thinner thread for surgery, and stiches for indestructable clothing. - Spider farms are not viable because the spiders are too aggressive to each other and production would also be too slow. - The gene for dragline silk was put into a goat and expressed in the mammary gland, resulting in the silk protein in the milk that could be spun into fine thread.

Describe applications within pharmaceutical biotechnology

- antibiotic production - human growth hormones - human insulin

What are some proposed methods of bioconfinement.

- conditionally express a trait. - sterility (ex, G2 seeds are formed and are fine for extracting oils out of them, but they will not germinate. This means that the G2 seeds will not grow and the farmer will have to buy more G1 seeds to produce more product} - lethality

Describe applications in agricultural biotechnology

- golden rice - FLAVR SAVR - arctic apples

Identify regulatory procedures involved with synthetic biology.

- it is unknown how to assess synthetic organisms for safety. - governments and companies have refrained from releasing these organisms into the environment because they might threaten the natural world. - the UN has called for caution and advisors and that none of these new organisms should be released into the wild at this time. - governments are unsure how to assess synthetically modified organisms for safety and indicate that they must be contained - Now SynBio organisms are being developed that are intended for release.

Describe other applications of biotechnology

- nanomedicine - gene therapy - stem cells - disease animal model systems - mammalian based drug production - fertility

Describe how penicillin acts as a bacteriostatic antibiotic. What types of bacteria does it target.

- penicillin was discovered by Flemming. It inhibits enzymes from forming cross-linkages in the peptidoglycan cell layer. This causes the osmotic pressure of the inside of the cell to be too great for the weakened cell wall to handle. As a result, the cell lyses. - penicillin V and G act against gram positive bacteria such as streptococcus and pneumococcus. - penicillin V and G are fermented products of the fungus, p. chrysogenum. This is a common fungus that is found naturally in moist soils with plentiful quantities of carbon and nitrogen.

What function do proteases play in the process of stain removal. How are they selected for and why is this important. Where do these proteases come from.

- proteases are good for cleaning blood stains and other soils caused by proteins. - alkaline proteases are obtained from Bacillus.

What function does cellulase play in deinking.

- used to increase paper strength.

What function do cellulases play in the process of stain removal. How are they selected for and why is this important. Where do these cellulases come from.

- when cellulases are used, it not only removes dirt, but it makes the colours more vibrant and the texture more smooth. - cellulases are obtained from genetically modified bacterial and fungi strains (Bacillus and Humicola}

Define disease animal model system.

-Animal models that are used in human disease research. - animal physiological systems are often so similar to humans that 90%of veterinary medicines are the same as, or very similar to, those developed to treat humans.

How is white-rot fungi used in the pulp and paper industry.

-White rot fungus, when grown in a pulp slurry with the addition of amylase, increases the brightness of the pulp. This doesn't decrease the pulp strength or yield. - this is a timely process. - the highest reported brightness accomplished by fungal treatment is 80% ISO (reflectance}.

How is spider silk pharmed.

1. A gene is removed rom the orb weaver and is attached to a segment of DNA that is called a promoter. This segment will turn on the gene when it is in the mammary glands of the goat. 2. A cell from a mother goat is taken and the nucleus is removed from the cell. 3. The spider gene and the nucleus are fused. 4. A fertilized donor egg is removed from another goat and the nucleus from the donor egg is replaced with the spider gene containing nucleus. 5. The egg divides a few times to ensure it will grow into a baby animal, and then implanted into a third mother goat. The final baby goat will carry the spider gene and pass it on when it has its own kids.

How is antithrombin pharmed.

1. Modifying the DNA - A human gene that produces the blood protein antithrombin is inserted into a short strand of goat DNA. 2. Implanting the DNA - The modified DNA is injected into the nucleus of a fertilized goat egg, which is then implanted into a female. 3. Testing the offspring - Kids born from the modified eggs are tested for the presence of antithrombin in their milk. promising kids are bred normally to create a herd normally to create a herd of modified goats. 4. Extracting the protein. Milk from the herd is filtered and purified. Annually, each goat can produce as much as antithrombin as 90,000 human blood donations.

What are the four components that make up GURTs.

1. Target gene which has a specific trait such as one that causes lethality (V-GURTs} or argonomic effects (T-GURTs} 2. promoter of the target gene that has a spatiotemporal patten (expressing the taget gene only in certain tissues and only at specific times} 3. Trait switch gene which is driven by a directly associated promoter, which reacts to signals from the genetic switch. 4. Genetic switch, which is an enabler gene and its associated promoter.

What are the advantages and disadvantages of using mice and a model system to study dieases found in humans.

Advantages - Mice have complex physiology that is more relatable to humans than flies do. - Metabolic processes are similar to humans. Disadvantages - Many animal model systems, for instance the use of rats and mice, may be sedentary, obese, or glucose intolerant. This may decrease their usefulness as a model for humans, as the metabolic processes are so different. - When observing psychiatric illness, there are concerns with qualitatice assessments of behaviour as they are often very subjective, leading to an experimenter observing what they want to, or expect to observe. - there are many ethical issues, such as how animal suffering is bad, etc.

What is an antibiotic.

- Antibiotics are biotechnological products that inhibit bacterial growth or kill bacteria. - they are naturally produced by microorganisms, such as fungi, at attain an advantage over bacterial populations. - bacteriostatic agents inhibit bacterial growth. This is so the immune system can take care of the infection. - bactericidal agents kill bacteria.

Discuss the ethics involved in antibiotic production.

- Doctors should only be prescribing antibiotics for bacterial infections, not viral infections due to antibiotic resistance. - What is an appropriate method for regulating who should be able to be prescribed an antibiotic.

Describe applications within industrial biotechnology

- Enzymes present in detergents - pulp and paper industry

Define society

Organized communities with shared laws, traditions, and values. individuals within societies, however, may not all share the same beliefs or live together. For instance, becasue of the internet, people are able to communicate with each other and live far apart, while still being part of a society.

How could you prevent contamination of a transgenic crop.

Reproductive control could achieve this through - reducing or eliminating gene flow - bioconfinement where genes are confined by a technology so that they cannot spread any further than defined by the technology

What are human recombinant protein drugs.

This includes insulin, human growth hormone, and blood factors.

What is an adjuvant

This is a fluid of lipid composition which acts to speed up the immune response. It is often added to vaccines.

Define morals

This is a personal standard of right from wrong.

What is a vaccine

This is a substance used to stimulate the production of antibodies and provide immunity against one or several diseases, prepared from the causative agent of a disease, its products, or a synthetic substitute, treated to act as an antigen without inducing the actual disease.

Define ethics

This is a system of moral principles recognized to a particular group or culture.

What are the pros associated with industrial biotechnology, what are the cons.

pros include - it uses renewable raw materials - it may lower greenhouse gas emmissions - lower petrochemical usage - has large economical potential - has large employment potential - may reduce negative impact on the environment (enzymes reduce chemical useage in detergents}. cons include - adding enzymes to detergents that babies chew on

How can we change an animals DNA.

1. Donor animal is given a hormone that causes her to produce many eggs. 2. Eggs are harvested. 3. Eggs are fertilized with donor sperm. Sperm becomes a pronucleus containing the father's DNA. 4. Transgene is microinjected into the male pronucleus. 5. Male pronucleus containing the transgene fuses with the egg nucleus. 6. Transgene moves into the egg genome. 7. Egg divides forming an embryo, and the transgene is duplicated in each new cell. 8. Egg is transferred to a surrogate mother's uterus to develop. 9. New animal that is born will produce the new protein product from the transgene, and will pass this ability onto its offspring.

What are the advantages and disadvantages of using herbacide resistant crops (farmer vs the industry}

Advantages - You can enable glyphosate to be used as an in-crop selective herbacide. - it provides easy, economical, efficient weed control. - it provides agronomic advantages such as earlier seeding and no tillage (tillage causes soil erosion}. - It has economic advantages as well. For example, US farmers save about 1.2 billion associated with the costs of herbacide purchases, application and tillage. For example, it allows US farmers to save about $385 million in tillage costs every year. Disadvantages to the farmers - farmers must purchase new seeds fom Monsanto each year becuase they are under contract. They must also purchase the Monasanto-specific herbacide (glyphosate}. - there is high reward for Monsanto, demonstrating the power of monopoly.

Describe how antithrombin III and tissue plasminogen activator are being used to treat blood clots.

Antithrombin and the tissue plasminogen activator are involved in preventing the anticoagulation cascade from getting out of hand.

What function do lipases play in the process of stain removal. How are they selected for and why is this important. Where do these lipases come from.

Function - lipases are involved in the hydrolysis of fats. Thus, they remove oil stains from fabrics. - lipases are used as major additives in laundry and household detergents. Selectivity - lipases have low substrate specificity, meaning that they can hydrolyse fats of various compositions - lipases have the ability to withstand relatively harsh washing conditions (pH 10-11, 30-60 degrees}, meaning you can use them in the cold wash. - lipases have the ability to withstand damaging surfuctants and other enzymes, which are important ingredients of many detergent formulations. - lipases obtained from p. glumae (bacteria} and r. mehei (fungus} remove food stains and sweat. They do this by converting water insoluble parts into water soluable parts (usually by converting triglycerides to free fatty acids and glycerol}.

What are sorts of methods are used within biotechnological applications.

Methods involve genetic engineering, cell tissue culture technologies, biochemistry, cell biology, genetics, microbiology, and molecular biology. You could modify living organisms, domesticate them, cultivate them, or make improvements to them through breeding programs that emply artificial selecteion and hybridization.

What regulations and legal aspects are involved in pharming. Compare Canada to the Us.

Regulations in Canada - Health Canada had authority on health products including drugs and biopharmaceuticles. - pharmaceutical products extracted from mammals falls under Health Canada's "Food and Drug act" - Guidelines and regulations for the health and wellbeing of animals used in biopharming activities fall under the Canadian Council on Animal Care and the "Health of Animals Act" Regulations in the US - Development, manufacturing, and distribution of medicinal products must follow Food and Drug Administration (FDA} regulations and guidelines. - The Center for Biologics and Research (CBER} has guidelines involving humans use, and The Center for Veterinary Medicine (CVM} has guidlines for animal use.

What are some ethical stances with regards to pharming.

The moral status of animals - Anthropocentrists believe that humans have the highest moral status. Animals and plants are instrumental to us, which means that it is okay to use them at our disposal when they can ethically be used for human benefit. - pathocentrists believe that having the capacity to feel is enough to deserve a moral status. Thus, some but not all animals are deserving of moral value. So somethings, like an individual cell, can ethically be used and disposed of for human benefit. - Biocentrists believe that all forms of life have value. Non-living entities havemoral value, but are valued because of their instrumental value to someone. - Holists believe that nonliving things have moral value due to their use by someone else as well as because they have intrinsic value. Naturalness - Doing "x" is morally problematic simply because it is unnatural. - playing god is wrong - there may be unprecedented risks of manipulating genomes (possible long term effects} due to verticle or horizontal gene flow. - introducing a foreign gene product (protein} from an animal into the human body can result in the transfer of pathogens.

Explain how mice are used to research ALS.

- Amyotrophic lateral sclerosis (ALS} is a rapidly progressive neurological disease which results in the gradual degeneration of motor neurons. It selectively attacks motor neurons in the motor cortex, brainstem, and spinal cord. - A large number of roden models are available for studying ALS - - The most important and most studied case of inherited ALS are mutations in the gene encoding superoxide dimutase 1 (SOD1}. SOD1 is one of the body's primary anti-oxidant defenses, and plays an important role in reducing oxidative stress in the body. It does this by partitioning damaging superoxide molecules into less reactive hydrogen peroxide molecules or regular molecular oxygen. - After the discovery of mutations in the SOD1 gene, a transgenic mouse overexpressing mutant SOD1 was created by inserting multiple copies of human genomic SOD1 into the mouse genome. - these transgenic mice show progressive hind limb weakness leading to paralysis and death. They replicate almost perfectly the disease process in human patients. - These transgenic mutant SOD1 mouse models were used to test new therapeutic strategies for ALS.

What are the ethical concerns of synthetic biology.

- Bioweapons - rapid construction of long strand of synthetic DNA also enables quick and inexpensive production of pathogens. - Monopoly - field of synthetic biology is already seen broadly worded patents. Only a small number of companies will gain monopoly control over economic sectors. - Affecting farming economy - SynBio can crash the farming economy (ex, saffron and vanilla}. - Decreasing biodiversity - farmers may resort to cutting down the rainforest (decreases biodiversity} in order to grow rice since the vanilla market could crash. - Violating nature - synthetic biology reconceptualizes life and organisms as being genetically programmable machines. - The Unknown - the creation of artificial life forms could have unpredictable impacts on the environment and biodiversity.

Who was Dover the Chimpanzee and how did he die. How dose this case study exemplify the differences between Canada and US in terms of animal care.

- Dover was a chimp being used in animal research. He died after Yerks national primate research center transported him in a cage hat failed to meet AWA regulations. - the cage was essentially a stainless steel box with solid flooring, roof, rear, and side, dangerous for confinment in high temperatures. - The temperatures inside the cage was too high under AWA regulations, and only the front of the enclosure that ventalation opening. - The USDA fined Emory University and the Yerks national primate research center 1,375 for violations of the AWA. - this case study exemplifies the small, somewhat insignificant fines associated with violating the AWA, as well as differentiations between Canada and the US in terms of animal care. - In canada, the ACC (animal care committees} can hault any research that is found to deviate from approved protocol or where the animals are found to be suffering excessive pain or distress, whereas the USDA is only able to issue penalties with facilities are not in compliance with AWA regulations.

Describe how erythropoietin EPO is being used to treat anemia.

- EPO is produced mainly in the kidney with little production in the liver. - kidney cells release EPO when oxygen levels are too low. - EPO promotes the formation of RBCs by the bone marrow. EPO also initiates the synthesis of hemoglobin (molecule that transports oxygen}. This acts to increase the oxygen carrying capacity of the blood. - EPO has often been misused as a performance-enhancing drug by some athletes.

Describe the current discovery scheme of antibiotics.

- Early stages of antibiotic discovery came from microbial product libraries. These libraries include antibiotics from environmental sources, such as soils, rhizospheres, plants, rocks, lichens, sediments and invertebrates. Antibiotics are then isolated, identified, and selected as part of a strain. Their sources are then fermented to produce large amounts of antibiotics. - After fermentation, the product undergoes screening procedures, compound isolation, and structural elucidation. - production is then scaled up. Compound development undergoes biological profiling, medicinal chemistry, and preclinical safety.

How are disease animal model systems used.

- Experimental - disease is induced artificially in the animal in the lab, and then possible treatments are sought out (most common}. - Spontaneous - disease occurs naturally in the studied animal, and treatment is sought ot (rare, but informative}. - Negative - refers to the control animals (there is nothing wrong with them}. This group is important for validating an experimental result. - Orphan - there is no human analog of the disease, and the disease only occurs in species studied.

How is GMO HGH produced.

- First, several synthesized DNA fragments were joined to a cloned fragment made from purified hGH mRNA. - Using enzymes, the hGH was added to circular DNA plasmids. - Newly constructed plasmids were introduced to a strain of E. coli. - Expressed hGH has been reported as identical to the natural substance (191 aa long}.

What is GenoCAD.

- GenoCAD is an open source computer-assisted-design (CAD} application for synthetic biology. It considers DNA as a language to program synthetic biological systems. - GenoCAD also includes design rules describing how parts should be combined in genetic constructs. - The same rules are used by the GenoCAD complier to maintain the integrity of existing constructs and artificial gene networks. - GenoCAD provides users with data import and export capabilities using standard formats so that users personal workspaces can be customized to meet their specific needs.

What are the types of disease animal model systems.

- Homologous models - share the same causes, symptoms, and treatment to a disease as humans. - Isomorphic models - have the same symptoms and treatments, but have different causes of the disease. - predictive models - are similar to human disease in only a few aspects but these may be significant and have importance in the medical field.

Why is pest management important for producing crops. Discuss prior methods of pest management.

- Insect pests have been been an integral problem of agricultural production worldwide. They lead to significantly lower yeilds of crops, and effect almost every aspect of the plants. - prior methods of pest management have used chemicals. - chemical pesticides results in environmental degredation (the chemicals can spread in the environment and effect beneficial invertebrates like honey bees}. In addition, you may have to spray a cropnumerous times in order to effectively kill a pest. Thus, a lot of chemicals could spread in the environmen, thereby increasing the harmful environmental impacts. - These chemicals can be harmful to human health and other organisms (carcinogenic}.

What are the ethics involved with disease animal model systems.

- It could be seen as ethically wrong to do any harm to an animal. Harm can be done through either experimental procedures or confinement in the laboratory. Some argue that by harming the animal, it can create inaccuracies in the study, and therefore yeild skewed results. - It could be seen as morally right to prevent or alleviate human suffering, which is why animal model systems are needed. - It could be seen as morally right to try to prevent innocent animals from suffering and being killed, which is why animal model systems are needed (we can save more through the sacrifice of a few}.

How is erythropoietin pharmed.

- It is pharmed in genetically manipulated chickens. - recombinant hEpo is produced in the serum and egg white of the GM chickens. - retrovirus is used as a vector to inject an expression cassette for a fusion protein of hEpo and the Fc region of human IgG into developing chicken embryos. - the oligosaccharide chain structure of the serum-derived hEpo is better than the egg-white derived hEpo.

What is a possible solution that could prevent Bt endotoxins from harming beneficial pollenators.

- Maybe you could find a promoter that reduces or eliminate pollen expression of Bt endotoxin. You dont need the endotoxin expressed in the pollen becasue it pests eat the plant, not the pollen.

Explain how mice are used to research HIV.

- Mice immune systems can carry key parts of the human immun system to replicate the human immune responce to HIV. This allows mice to be used to test HIV in vaccine canidates. - BLT (bone marrow/liver/thymus} mice were engrafted with human bone-marrow cells, liver and thymus tissue. They produce a functional human immune system with CD4+ T cells (the target of HIV}. BLT mice also express HLA-B57, a human gene variant that protects against HIV, and show better control over HIV replication. During the acute phase of infection, these mice generate HIV-specific killer T cell responses that closely resemble those in humans. However, the virus rapidly evolves to escape these responses (just as it does in humans}.

Who is Monsanto. What is their product. How was it made.

- Monsanto is a multinational agrochemical and agricultural biotechnology corporation. They are the leading producer of the herbacide glyphosate, and of genetically modified seeds to glyphsate resistant crops. - Monsanto markets their glyphosate resistant crops under the Roundup Ready Brand. Their product - Glyphosate - - glyphosate allows farmers to kill weeds without killing their glyphosate resistant crops. - it is considered safe because it does not break down into toxic compounds apparently. - glyphosate is non-selective - glyphosate inhibits aromatic amino acids from being synthesized, inhibits protein/secondary product biosynthesis and results in deregualtion of the shikimate pathways. All of this leads to general metabolic disruption. Their product - Roundup Ready - The glyphosate resistant crops were created by stable integration of a transgene using Agrobacterium tumefaiens mediated transformation. This transgene only makes the plant glyphosate resitant, it does not affect the crop in any other way. How the crop was created - - C4 strain of Agrobacterium was found growing in wastefed column at a factory that made glyphosate. The bacteria had a gene that encodes for glyphosate-insensitive 5-enolpyruvylshikimate-3-phosphate synthase (epsps}, which is the target enzyme for glyposate in the shikimate pathway. The epsps was almost completely insensitive to glyphosate. - The C4 epsps bacterial gene was cloned and inserted into a bacterial plant vector (plasmid} in order to prepare for cloning into plants. It was modified to ahve a strong plant promoter. - the shuttle plasmid was build in E. coli then purified plasmid DNA was used to transform Agrobacterium tumefaciens. Agrobacterium tunefaciens infects plants and injects DNA from a Ti-plasmid into plant cells. This results in DNA entering the nucleus of the plant and being incorporated into the plant chromosomes. - With the transgene integrated into the plant's genome, you could then spray the plant with glyphosate and it will do okay. - Corn, canola, tobacco, and cotton all have been engineered to be resistant to glyphosate. - Monsanto's patent on the glyphosate-resistant crop technology expired in September of 2000. However, they still dominate 80% of the market share of the glyphosate-based herbacides.

Compare GMO insulin to its natural counterpart. What ethics (benefits and disadvantages} are involved in the production of GMO insulin.

- Natural insulin is produced by pancreatic cells. To treat diabetes, insulin from pancreatic cells of mammals can be used. - GMO insulin is produced by E. coli in 50,000 liter fermenters. - A batch of insulin from one fermenter could produce a year's supply of insulin for thousands of people. - benefits of GMO insulin - microbes reproduce more rapidly and with fewer resources than complex mammals, lower cost, reduces delay associated with the animal maturation, space requirements are relatively low, the supply is high and does not depend on the quantity or availability of bovine or porcine pancreases, and finally GMO insulin is "identical" to natural human insulin. In contrast, there may be slight differences in mammalian derived insulin. Thus, it should work like normal insulin in your body. - disadvantages of GMO insulin - GMO insulin poses a risk to some patients with specific genetic backgrounds. This person can make an anti-insulin response. The society of diabetic rights found that a significant number of diabetics have experienced bad reactions to GMO insulin, and some of these has resulted in death.

Describe how vaccination can be potentially used to target West Nile Virus.

- RNA is extracted from the West Nile virus and converted into DNA. - the genetic sequence for the west nile virus is generated from the DNA. - based on the DNA sequence, primers specific to the prM and E gene region are produces. These primers will be used to generate cDNA fragments containing both prM and E genes. - the cDNA fragment is inserted into a plasmid - the plasmid carrying the genes are grown in large quantities in the bacteria and purified by column chromatography. - the purified DNA plasmids carrying the prM and E genes make up the investigational vaccine.

Compare synthetic saffron to its natural counterpart. What ethics are involved in the production of synthetic saffron.

- Saffron is usually made from the crocus flower in Iran. - Evolva, a Swiss-American SynBio company, has re-engineered yeast to contain the metabolic pathway for production of key saffron compounds. The yeast expresses saffron-derived genes in engineered microbes. - Evolva can now brew their synthetic saffron in a large vat of engineered yeast similar to the production of beer and wine. - Ethics involved - every hectare of natural saffron growing in Iran provides jobs for 270 people per day. By replacing saffron with synthetic saffron, you threaten those jobs.

What are some approaches that can be used to improve antibiotics.

- Strain improvement can either be random or rational. - Random methods include- optimization of culture conditions and random mutagenesis and further screening. - Rational methods include- bioinformatic-guided engineering based on omics data and genome-scale metabolic network models, heterologous expression of clusters in surrogate hosts, amplification of clusters, manipulations with primary metabolism (flux of precursors or sigma factor or ribosome engineering}, manipulations with structural or transporter and resistance genes in the cluster, or through manipulations with global and cluster regulatory genes (over-expression of positive regulators using native or heterologous promoters, or inactivation of negative regulators}.

What are some America laws and regulations that relate to the use and treatment of animals in research.

- The Animal Welfare Act (AWA} is the only US federal law that covers animals in research. - It regulated the care and use of animals in research, testing, teaching, exhibition, transport, and by dealers. - The AWA provides minimal protection for certain species, while excluding others such as rats, mice, and birds bred for research. The AWA also excludes cold-blooded animals (fish, amphibians, reptiles} and farmed animals raised for food, fibre, or agricultural research. - The US Department of Argiculture (USDA} is responsible for enforcing the AWA. The USDA's Plant Health Inspection Service (APHIS) Animal Care program administers AWA regulations and standards. - Under the AWA, buisnesses and individuals using regulated animals must be licensed or registered with the USDA and facilities with regulated animals must be inspected yearly by APHIS. - Under the AWA, research institutions are required to establish an Institutional Animal Care Use Committee (IACUC} to oversee and evaluate all aspects of the institution's animal care and use program. The animal facility's CEO selects all the IACUC members, so research facilities are self-regulating. - There is no legal requirement for the inspection of federally-owned and operated research facilities. - In addition, the USDA has no jurisdiction over facilities using animals not covered under the AWA. - Furthermore, when the USDA fines facilities for non-compliance to AWA regualtions, they may issue penalties up to 10,000$. These fines are typically very small though.

What is the CCAC and ACC and what do they have to do with regulating aspects of animal model systems.

- The Canadian Council on Animal Care (CCAC} guides all aspects for the care and use of experimental animals. The program has brought about high standard for experimental animal care and use through education, voluntary compliance and a code of ethics. - The CCAC has developed "The Guide to the Care and Use of Experimental Animals," in which participating facilities must adhere. - To carry out its mandate, the CCAC adopted a decentralised model in which responsibility for all matters relating to the care and welfare of experimental animals resides within the institution, through the animal care committee (ACC}. - The ACC in each institution has a number of responcibilities relating to the care and use of animals in research, testing, and teaching. - The ACC must ensure that every proposal to use animals for teaching, testing or research has been viewed both for scientific merit and for compliance with accepted ethical standards. - Each protocol must be subject to annual review, and any amendments reviewed and approved. - The ACC has the authority to halt any study that deviates from teh approved protocol, or any study where the animals are found to be suffering excessive pain or distress that cannot be relieved. - The ACC must also ensure that facility standards and the care of animals are in accordance with CCAC guidelines, that adequate veterinary care is provided, and that all persons working with animals recieve proctical skills training.

What are some provincial regualtions and laws related to the use and treatment of animals in research.

- The ontario legislation, namely the Animals for Research Act, is unique in Canada in that it creates a system of control based on the registration of the research facilities and the issuance of licenses of supply facilities. - All research facilities that use animals in their work must be registered.

What are conditional lethality factors used for.

- These factors are used to control transgenic volunteers. - An example of a conditional lethal system is one that uses IAAF gene from Agrobacterium. IAAH encodes indoleacetmide hydrolast which convets IAM to the auxin IAA. - the inclusion of an auxin transport inhibitor maximizes the effec of toxic auxin production - too much auxin causes rapid cell growth. - the plant dies when their transport inhibitory maximizes the effect of the toxic auxin production.

How do proteinase inhibitors act to control pests. What are some examples.

- This is another insect resistant mechanism. - proteinase inhibitors are natural, defense-related proteins often present in seeds and induced in certain plant tissues by herbivory or wounding. They act to increase the plant's defense arsenal. - proteinase inhibitors are present in multiple forms in numerous tissues of animals, plants, and microorganisms. Examples - proteinase inhibitor factor (pif} that switches on a cascade of events. - this leads to the synthesis of inhibitor proteins. If an organism does this, then the insect consumes the inhibitors and it cannot digest the food it eats. The insect will either die or leave, thereby protecting the plant.

Describe the controversy between Monsanto and Schmeiser.

- To start, this was a case based on patent rights, which occurred between August 1998 and January 2004. Monsanto developed a GM crop, called Round Up Ready Canola, which was patented. - Schmeiser never paid Monsanto to use their RRC. - In 1998, test revealed that 95-98% of the crop that Schmeiser was growing was RRC. Schmeiser suggested that the reason the RRC was being grown in his field was because that the seeds fell of a cart and went into his field. - The Canadian Court found Shcmeiser guilty of "selling or otherwise deriving the plaintiffs of their exclusive right to use plants which the defendants know or ought to know are Roundup Tolerant, or using the seeds from such plants. - Monsanto was given the right to retrieve their patented genetic material. - Schmeiser was ordered to pay Monsanto damages. - The Appeals court upheld this ruling. - Schmeiser then appealed to the Supreme Court. Its ruling upheld the validity of Monsantos's patent. However, the trial judge stated that "none of the suggested sources could reasonably explain the concentrations or extent of RRC of a commercial quality that was ultimately resent in Schemiser's crops. However, Schmeiser did not owe Monsanto either damages or court costs. The court stated that in a case like this, the amount of damages is measured by the extra profits derived from the use of the patented item. Since Schmeiser did not spray any of the herbicides, he did not enjoy any of the benefits from it.

What are some federal regualtions and laws related to the use and treatment of animals in research.

- Under the constitution act, the federal government does not have jurisdiction to legislate with respect to experiments involving animals as this is provincial jurisdiction. - However, the federal government is not totally absent. 3 areas where they can take action are under the criminal law power, the health power, and the spending power. - The criminal code of Canada protects animals from cruelty, abuse and neglect. - The health of animals act are aimed at protecting livestick from a variety of infectous diseases that would threaten both the health of animals and people, and Canadian trade in livestock with other countries.

Define industrial biotechnology and some of its applications

- Utilization of biotechnological processes in industrial production - Making chemicals, food, detergents, paper, pulp, textiles, biofuels. - development of new products, processes and services, as well as improving existing ones. For instance, this could be done using synthetic biology. You could improve detergents, the deinking process, you could make spices. You could improve the pharmaceutical industry.

Compare synthetic vanillin to its natural counterpart. What ethics are involved in the production of synthetic vanillin.

- Vanilla is naturally found as part of the orchid family. - Yeast was genetically engineered with computer generated DNA, causing the yeast to synthesize vanillin. - it is unclear if synthetic vanillin is safe to consume (i.e. it is unclear how synthetic organisms interact with natural organisms or ecosystems}. Thus, scientists may not find urgent health threats until it is too late. -Ethics involved - An estimated 200,000 people grow vanilla beans in Madagascar, Uganda, Mexico, and elsewhere. SynBio vanillin could significantly affect farmer's business income. This could threaten the ecosystems because vanilla farming is closely related to rainforest protection (natural vines require intact forest to thrive}. If the price of natural vanilla crashes due to SynBio, farmers may resort to deforestation and plant rice to survive.

Describe how drosophila melanogaster is used to treat Alheimer's disease.

- We use fruit flies because they are easier to construct over transgenic mice. - In humans, the overexpression of Tau proteins causes age-dependent neurodegeneration, axonal transport deficiets, and early death. These are all signs of Alzheimer's disease. - Transgenic flies that address Tau or amyloid oxicity have been developed to study Alheizmer's disease.

How is synthetic insulin produced.

- You start with a half a gram of E. coli. You give them a source of macro and micro nutrients in order to grow. You then make the bacteria ampicillin resistant to prevent other organisms from invading the fermenting process. During growth, you repress the bacteria from making insulin. - An inducer is used to start insulin production. You start with two chains- A and B- to allow for the manufacturing of each of the specific peptides that are required. - Small coding sequences are synthesized for polypeptides A chain (21aa} and B chain (30aa}. These are then transformed into the E. coli. - the promoter B galactosidase gene is fused to insulin A fusion protein gene. - After being transformed into the cell, the B galactosidase and insulin A fusion protein accumulates within the cell. These are extracted and treated with cyanogen bromide, which acts to cleave the A and B chains. Next, you purify the chains and then mix them together (under certain conditions}. Disulfide bonds form through an oxidation reaction which joins them together to give rise to the functional form of insulin.

What function do amylases play in the process of stain removal. How are they selected for and why is this important. Where do these amylases come from.

- alpha-amylases remove starch oriented stains such as those caused by chocolate, cocoa, and puddings. - beta-amylases and iso-amylases catalyse hydrolysis of glycosidic linkages in starch. - these are obtained from genetically modified Bacillus and some fungi. - there is potential for amylases to destroy the environment.

Compare GMO Blood clotting factors (8 and 9} to their natural counterparts. What ethics (benefits and disadvantages} are involved in the production of GMO Blood clotting factors.

- blood clotting factors are used to treat patients with hemophilia. - natural blood clotting factors were obtained from donated blood. However, donated blood was inadequately screened for HIV, thereby posing a significant risk to patients with hemophilia who received these blood clotting factors from donated blood. This eventually caused them to develop AIDs. - GMO blood clotting factors are produced using recombinant DNA technology and mammalian models. - Benefits of GMO blood clotting factors include - patients will not develop HIV/AIDs, they work. - Disadvantages of GMO blood clotting factors - Jesse, what if these factors are expensive and someone cannot afford treatment, not everyone may be able to receive treatment due to allergies to the mammals that are used to produce these drugs.

What function does lypase play in deinking.

- catalyzes the hydrolysis of ester chemical bonds - acts on liposoluble compounds.

What function does laccase play in deinking.

- degrades lignin - causes detachment of the ink particles linked to lignin - it is a copper-based oxidative enzyme that reacts with phenolic residues of lignin in the presence of oxygen as an electron acceptor. - a synergistic deinking effect is created with both cellulase and hemicellulases (wood is composed of this and lignin} and laccase system. - the enzyme-combined deinked pulp achieves a higher brightness -

What function does xylanase play in deinking.

- degrades the linear polysaccharide beta-1,4-xylan to xylose, thus breaking down hemicellulose. - a mixture of cellulases (improves paper strength} and xylanases is optimal for treating laser printed material or newspapers.

What role do enzymes take in the process of stain removal and the deinking process

- enzymes remove stains from clothes and ink from paper. - enzymes reduce the need to use chemicals in detergents. - lipases are involved in the hydrolysis of fats, removing oil stains from fabrics. - cellulases remove dirt from fabrics, makes colours more vibrant, and texture smooth. - amylases removes stains from chocolate. - proteases remove blood and other soils caused by proteins.

How do you acquire enzymes for industrial biotechnological applications

- fermentation of bacteria (could be genetically modified} under optimal living conditions produce enzymes for industrial use. - the enzymes are isolated and further processed after fermentation (cleaned to remove certain materials that are bound to them}. - Novo Nordisk and Gis Brocades are two manufacturers of enzymes which are used in detergents.

What problems can be addressed by biotechnology

- food crisis - industrial processes - alternative fuels - medicine and vaccinations - soil and water regulation

Compare GMO hGH to its natural counterpart. What ethics (benefits and disadvantages} are involved in the production of GMO HGH.

- hGH acts to mediate both growth and stature. When the pituitary gland secretes insufficient amoung of hGH, it can result in hypopituitary dwarfism. As a result, hGH is used as a treatment. Although this is a rare disease, hGH supplies are limited. - hGH used to be obtained from the pituitaries of human cadavers. - GMO human growth hormone is produced by E. coli created by Genentech Inc. - Benefits of GMO hGH include- creates a steady supply of hGH whereas without it the supply would be scarce, and the expressed hGH is identical to the natural substance. - Disadvantages of GMO hGH include - maybe not everyone can receive it.

Describe applications of synthetic biotechnology

- herbicide resistant crops - pollen control - insect resistant crops - virus resistant crops - preformance plants

Briefly describe the process of synthetic biology. What is Synthia.

- synthetic biologists use a DNA printer and DNA synthesizer. - it builds artificial DNA from scratch to arrange the DNA in any way. - you don't need DNA cloned from nature anymore (because there is a DNA printer}. - it is even possible to synthetically print out from scratch all of the DNA of a living organism. - In 2011, Craig Venter created a microbe, nicknamed Synthia, whose full set of DNA had been printed out by a machine. He called it the first self-replicating species on the planet whose parent is a computer.

Define synthetic biology. Give examples of products created from SynBio.

- synthetic biology is often referred to as extreme genetic engineering and still remains highly unregulated. It is a form of genetic engineering that adds manufactured genetic parts (such as synthetic DNA, or RNA} into a living cell. - scientists can write entirely new genetic codes on a computer, print them, and insert them into living organisms. Researchers attempt to program the code of DNA in order to alter the behaviour of the organism. - synthetic biology allows the design of DNA sequences that have no known analogue in nature. For instance, some researchers are inventing entirely new types of DNA composed base pairs that are not found in nature. - products that have been made from synthetic biology includes- vanilla substitute grown by synthetically modified yeast, coconut oil replacement produced by engineered algae, and patchouli and vetiver fragrances.

How is alpha-1-antitrysin (ATryn} being used to treat emphysema and CF.

- the ATryn rDNA construct was put into goats so that they would produce ATryn. - ATryn is used to treat patients with antithrombin deficiency.

Compare the conventional deinking process to the biodeinking process.

- the conventional deinking process uses caustic soda, silicate, and peroxide to remove ink from recycled papers. However, these techniques are only useful to deink oil-based printing materials (news papers and magazines. Thus, conventional methods are insufficient for the production of high quality pulps. - currently, there are new coatings and new types of inks containing synthetic polymers in laser and xerographic printing. The ink that adheres to these surfaces can be removed if treated with enzymes that attack surface components. Such enzymes include- lypase, laccase, cellulase, and xylanase. - lypase catalyzes the hydrolysis of ester chemical bonds, acting on liposoluble compounds. - laccase degrades lignin, causing detachment of the ink particles linked to lignin. - cellulase improves paper strength. - xylanase degrades the linear polysaccharide beta-1,4-xylan to xylose, thus breaking down hemicellulose. - a mixture of cellulases and xylanases is optimal for treating laser printed material or newspapers.

Why is bioconfinement of transgenes a topic of vigorous debate.

- there is concern if the crop and wild type relative can cross, and what would happen if it does (i.e. the wild relative could end up in bioconfinment}. -There is concern when transgenes and their conferred traits, such as drought, salt or cold resistance, are not completely understood with regards to fitness. - there is concern if crops producing pharmaceuticals require bioconfinement (i.e. should this be open for everyone's use} - thus, instead of bioconfinement, technologies that focus on eliminating gene flow would be beneficial.

Describe how amoxicillin and ampicillin act as bacteriostatic antibiotics. What type of bacteria do they target.

- these can be used to target gram negative bacteria, such as E. coli and H. influenza. - they work by inhibiting crosslinking of linear peptidoglycan polymer chains. This eventually leads to osmotic lysis of the cell, thereby killing the bacteria.

What are herbacide resistant crops

- these fall under synthetic biotechnology - these are crops that are naturally or have been genetically selected or modified to be resistant to herbacides. - these crops contain genes that enable them to degrade herbacides or lack herbacide targets.

What makes bacteria and fungi effective for industrial use

-Bacteria are effective because they are easily cultured (using yeast, agar, plating tecniquies, have fast growth (doubling growth is about 20 minutes}, they are inexpensive, replicable, easily maintained and contained, as well as easy to genetically modify in order to produce all different kinds of enzymes. - Fungi are also effective for industrial biotechnology because they are a source of unique gene products, easily cultured, fast growing, and transformable.

Name 7 types of vaccines.

1. Live, attenuated vaccines 2. Inactivated vaccines 3. Subunit vaccines (pieces of the actual pathogen} 4. Toxoid (detoxified toxin} vaccines 5. Conjugate vaccines (use linked bacterial surface polysaccharides to other antigens} 6. Recombinant vector vaccines (antigen genes in attenuated infecting viruses or to be expressed on the surface of harmless bacteria} 7. DNA Vaccines (human cells can take up DNA and make foreign antigens that are exposed to the immune system}

What are the advantages and disadvantages of using Drosophila as a model organism to study diseases found in humans.

Advantages - No ethical problems/no restrictions according to protection laws. - easy and cheap to maintain in large quantities, time and cost effective handling. - genetic manipulation is fast and inexpensive (3 months, and under 500$ per transgene}. - there is a plethora of available resources/stocks (e.g. genome-wide RNAi-library} - they have a short generation time (10 days}, short lifespan (2-3 months}, and are easy to use for screens. - they have a fully sequenced and annotated genome. - they have good conservation of basic signalling pathways and cellular processes in general. - they have low redundancy/reduced number of paralogous genes compared to vertebrates - they are one of the best understood multi-cellular organism. - they have balancer chromosomes which allow the maintenance of mutations/transgenes without genotyping. Disadvantages - Their brain anatomy, cardiovascular system, and respiratory system differs substantially from humans. - they have no easy measure of complex behaviour - they have only basic measures of cognitive decline - they sometimes have poor conseration of proteins/protein function - they are maintained as living creatures only, with no permanent conservation (i.e. freezer stocks are not available}. - they have a less complex and adaptive immune system than in vertebrates. - effects of drugs on the organism might differ strongly compared to humans (e.g. conversion of protoxins in the liver}.

What are the advantages and disadvantages associated with using Bt-crops.

Advantages - Reduces the need for conventional insectiides which benefits human health and the environment. - Bt corn has been shown to have no effect on honeybees or beneficial predators. Honeybees to not gather pollen from corn, so the modified BT corn does not affect them. This means you can be a part of integrated-pest management (helpin beneficial insects while killing bad ones}. Disadvantages - Many varierties must be used if you want to fully protect your crops. This is needed in order to provide protection from many different insects. - Ineffective becasue some insects such as the codling moth dont get a change to eat it. This means there is a chance to become resistant against Bt crops and then it could be useless. For instance, this happened with cotton. Insects can detect when something is not the best thing to eat and then they go to the refugia (crops that are not pest resistant}. - When the Bt corn pollen is dispersed, it lands on other plants, such as milkweed, which is a food source for Monarch butterfles (very imporant pollinators}. pollen from Bt corn could cause a serious risk to populations of monarch butterflies and other butterflies, especially since the corn plants can produce a lot of pollen. In fact, lab tests show that Monarch butterflies grow more slowly and suffer higher mortality rates when the milkweed leaves are contaminated with the BT endotoxin (the transformed pollen goes into the gut of the catepillar, where it binds to specific sites}.

What are the advantages and disadvantages of V-GURTs.

Advantages - increase productivity from improved imputs due to increased research and development investment. Disadvantages - it makes farmers dependent on external seed sources - it violates the rights of farmers to save and reuse their own seeds - it adds an additional cost to the farmers - could have negative environmental impacts such as resistance of other organisms to the variety produced.

What process is used to create herbacide resistant crops.

Agrobacterium tumefaciens mediated transformation - this is the most commonly used method for plant genetic engineering which has relatively high efficiency. How it works 1. The pasmid is removed from the bacterium, and the TDNA is cut by a restriction enzyme. 2. Foreign DNA (the desired gene} is cut by the same enzyme. 3. The foreign DNA is inserted into the TDNA of the plasmid. 4. The plasmid is reinserted into a bacterium. 5. The bacterium is used to insert the T-DNA carrying the foreign gene into the chromosome of a plant cell. 6. The plant cells are grown in culture. 7. A plant is generated from a cell clone. All of its cells carry the gene and may express it as a new trait.

Describe 3 approaches of how vaccines can be developed to target microbes.

Approach One - through genetic engineering, scientists can isolate specific genes and insert them into DNA of certain microbial or mammalian cells. - these living factories mass produce the desired antigen. - using another products of biotechnology, a monoclonal antibody that binds to the antigen, the scientists can separate the antigen from all other material produced by the cells. - this technique has been used to produce immunogenics of the hepatitis B virus and the malaria parasite. Approach Two - insert genes for desires antigens into the DNA of the vaccinia virus, the large cowpox virus - when the reengineered vaccinia virus is inoculated, it stimulates an immune reaction to both the vaccinia and the products of its passenger genes. - these have included, in animal experiments, genes from the viruses that cause hepatitis B, influenza, rabies, and AIDs. Approach Three - scientists isolate the gene that encodes an antigen and determine its amino acid sequence. - they then identify small important areas of the larger protein molecule, and synthesize it chemically. - wholly synthetic vaccines are being explored for malaria and for the major diarrheal diseases that are affecting developing countries.

What are the benefits and drawbacks of using animals or cells for pharming.

Benefits - the animals/cells are reproducible, have flexible production, and are easily maintained. Also, acquiring milk from them to use as a system is easy. - since the mammary gland and milk are not part of the main life support systems of the animal, there is little risk of harm to the animal that is making the transgenic protein (because it is designed to be in the milk and then it is secreted away... it does not build up within the animal}. - Even though the transgene is present in every cell in the body, it is only active in the mammary gland due to the use of a specific promoter. Drawbacks - produce low yields of the pharmed product. - cultured mammalian cells are sensitive to shear forces that occur during industrial scale culture, and to variations in temperature, pH, oxygen, and certain metabolites (a single cell cannot make its own metabolite, so you need to feed it this}. Thus, cultured conditions need to be controlled carefully. - the requirement of having fetal bovine serum in the growth medium makes production expensive.

What controversies exist relating to the production of herbacide resistant crops.

Herbacide resistance - weeds could be selected through spontaneous mutation to be herbacide resistant, making the product useless. - for instance, the use of glyphosate resulted in giant ragweed being resistant. This weed is about 8 feet tall and its pollen is really nasty to people. Because they are resistant, they are now growing in canola pathches and are hard to control. - in order to prevent weed resistance to herbacides, it is recommended that farmers rotate Roundup ready crops with other crops, as well as use alternate herbacides. Seed contamination - through a variety of means, Roundup ready genes have been introduced into the food supply. - also virtually impossible for farmers to grow non-genetically engineer Canola in Canada. Lack of yield increase - Monsanto claims to be addressing the needs of the word when it comes to the food supply. However, there is no evidence that the use of Roundup ready crops increases the yield of farmers who use their seeds. - GM crops are developed to be pesticide and herbacide resistant, rather than directly increase the yield of a given crop. Halted sale of Alfalfa seeds (Monsanto tried to jump the gun on regulations here} - In 2006, Roundup ready Alfalfa was commercialized, and the Center for Food Safety sued the USDA for failing to prepare an Environmental Impact Statement (EIS} on the product. - In 2007, a District Court stopped the sale and planting of Roundup Ready alfalfa see, and in 2011, after completing the EIS. the USDA again approved sales and planting. Toxicity - there is speculation that glyphosate can increase the chances of breast cancer cell proliferation. - it is also speculated that glyphosate may cause intestinal and gut damages contributing to diabetes, G.I. disorders, heart disease, obesity, autism, and parkinsons. - glyphosate is slightly toxic to birds and fish, and it binds tightly to the soil, reducing the possibilities of leaching. - microbes in the soil then brak down glyphosate so it does not accumulate. - if glyphosate is broken down, it is thought that it is not likely to have negative effects to humans.

Compare conventional methods and biobleaching methods that are used to make paper white.

The pulp and paper industry uses enzymes to chemically process various types of wood pulp to decrease the colour of the pulp. The main use of the wood pulp is to make white paper. - the conventional method uses bleaching with chlorine and chlorine dioxide in order to produce bleached paper pulp. This is then used to make white paper. - biobleaching involves prebleaching with cellulase-free xylanases (this reduces environmental impact}. This allows for bleaching to occur with much less chloride compounds in order to make bleached paper pulp. This is then used to make white paper.

What is pharming.

This is mammalian based drug production through genetic engineering of an animal. It can either use the animal itself or its cells.

Compare and contrast SynBio to transgenic organisms.

Transgenic - DNA is exchanged between species. Synthetic - These organisms are a different type of life form. - DNA is not extracted from a living organism, but is built from scratch.

What are Bt-crops. What process is used to create them. How do they effect insects.

What are Bt-crops - Bt-crops are trasngenic crops which contain the Bt toxin gene - there are thousands of different Bt strains, producing over 200 types of these proteins. - Bt corn and Bt cotton was grown on 22.4 million hectars worldwide in 2004 How are they produced - The soil microbe Bacillus thuringiensis (Bt} is a gram positive sporulating bacterium. It produces proteinaceous crystalline inclusion bodies during sporulation. - the Bt toxin gene of the bacterium was transferred to trangenic crops. First, the Bt gene that produces the desired lethal protein was joined to a marker gene for antibiotic resistance. Next, the Bt gene and marker gene are inserted into plant cells. The plant cells are grown in the presence of the antibiotic. Cells carrying the Bt gene and antibiotic resistance gene survive and are grown into plants. How they effect insects - The crystal proteins provide host plant resistance to major pests. - they are highly insecticidal at low concentrations - crystaline inclusion bodies (Cry proteins and Cyt proteins} have different targets in the insect's guts. - the insects eat the Bt crystals and spores. - the toxin binds to specific receptors in the gut. - the crystals cause the gut wall to break down, allowing spores and normal gut bacteria to enter the body. - the insect dies as spores and gut bacteria proliferate inside its body.

What are GURTs. Explain their use in industry created crops.

What is GURTs - GURT refers to Gene Use Restriction Technologies. - It was originally developed for intellectual property protection. What is V-GURTs - V-GURTs stands for variety-specific gene use restriction technologies. - they restrict at the plant variety level. - these technologies produce sterile seeds that cannot be used to plant a crop. For example, it you target a trait, like gene expression of seeds, you can prevent seeds from germinating unless provided with an inducer chemical available only to the seed company. Here, the entire genome of the plant may be eliminated from the seed. - use this technology to protect products from unauthorized use What is T-GURTs - T-GURTs stands for trait-specific gene use restriction technologies. - they resrict at the plant trait level. - these technologies produce a crop that does not function until the crop plant is treated with a chemical. The target gene results in a particular trait (such as herbacide resistance}, but not lethality. Thus, when T-GURTs are applied to crops, required purchase of the activator compound is necessary in order for the farmer to take full advantage of the crop.

Discuss what you learned about hemophilia from the guest lecture.

What is hemophilia - hemophilia effects the hemostatic system, which is e balance between bleeding and clotting. - in hemophilia, you are missing the active form of enzyme factor 9, and/or cofactor 8. Hemophilia A is factor A deficiency, and Hemophilia B is cofactor 8 deficiency. Hemophilia B is more common. - when missing these components, hemophilia causes spontaneous bleeding problems. This bleeding can go into joints, causing destruction. - genes that encode factor 9 and cofactor 8 are found on the X chromosome. - the gene for factor 9 is made up of 8 exons, and the gene for cofactor 8 is made up of 26 exons. Note, two mRNAs are transcribed from the cofactor 8 gene- FaA and FaB. - Queen Victoria was a carrier for hermophilia. How do you treat this condition - you could replace the missing protein via frequent repeated intravenous infusions. The proteins are unstable and have short half-lives so they have to be done via intravenous infusions (a couple of times a weeks} - the clotting factor source to treat this condition comes from plasma donations. - Other sources of clotting factors come from recombinant DNA production. Recombinant proteins are made, concentrated, adn infused into the patient. Most patients in Canada are treated with these recombinant proteins, but this is very expensive. Countries that cannot afford this method are still using plasma donations and the plasma is screened for viruses. Note- making and concentrating the recombinant protein is difficult. - new innovations include cofactor 8 antibody mimicry. To make the clotting complex work, you need factor 9, cofactor 8, and substrate 10. When missing 8, the complex does not work because you cannot bring it all togehter. By using a cofactor 8 antibody, you can bring together factor 9 and substrate 10 in an efficient fashion. This antibody is not as good as the actualy cofactor 8, but it is a major advance. The good thing about this therapy is that the antibodies work for weeks, not just days (compared to the other therapy described}. - another innovation is through using gene therapy. Here, you use viral vectors to deliver genes to the nucleus of somatic cells. You use viral vectors because they have evolved for gene transfer. Researchers have been using adeno-associated virus and various forms of retro-viruses for this therapy. In this therapy, the gene Rep and Cap are replaced by cofactor 8 cDNA and a promotor using the virus. Note- Queens university tested this on dogs with Hemophilia B (facotr 9 deficiency}, and found that after intravenous injections, the dogs have a decrease in spontaneous bleeding throughout the year. - these new treatments can have implications for the old technologies. The old technologies are very expensive and have to be continuously injected. If you could cure the disease with just one injection, then that would place panic on the old technologies (money}.