BASIC SCIENTIFIC OF BIOTECHNOLOGY What Is Biotechnology and

BASIC SCIENTIFIC OF BIOTECHNOLOGY

What Is Biotechnology and What Does It Mean to You? • Biotechnology – using living organisms, or the products of living organisms, for human benefit to make a product or solve a problem • Historical Examples – Fermentation – Selective breeding – Use of antibiotics

• Example of Biotechnology – Selective Breeding (a) (b) Normal zebrafish • "Casper" zebrafish – made by selective breeding What feature of Casper makes it a "model organism" to study migration of cancer cells compared to wildtype fish? Via selective breeding, ﺍﻻﻧﺘﻘﺎﻟﻲ ﺍﻟﺘﺰﺍﻭﺝ by mating a zebrafish mutant that lacked reflective pigment with a zebrafish that lacked black pigment, Casper is transparent. This feature allows scientists to inject fluorescent cancer cells into Casper's abdominal cavity and then track the migration of those cells to specific locations in his body.

• Based on this tree, can you become successful in the biotech industry only studying biology? No, you need to take many science courses including biology, computer science, chemistry, math (especially statistics). If you are not broadly trained, you will not be able to obtain a job in this growing field!

• Modern Examples – Gene cloning – Genetic engineering – Recombinant DNA technology – Human Genome Project

• Example of "modern" biotechnology: – recombinant DNA technology started modern biotech as an industry • Examples of applications – – • development of disease-resistant plants food crops that produce greater yields "golden rice" engineered to be more nutritious genetically engineered bacteria that can degrade environmental pollutants Gave more examples of applications ؟ : . -Low cost production of proteins used to treat diseases -Development of recombinant enzymes to be used by molecular biologists - for research

• Look at the two chromosomes and determine which chromosome has more than one gene involved in promoting breast cancer. Explain your answer. – Now use the link to further study the diseases involved in these chromosomes • http: //ghr. nlm. nih. gov/chromosome 13 has 2 genes listed regarding breast cancer and chromosome 21 only has 1 gene related to breast cancer.

• • Most drugs are developed to combat ﻣﺤﺎﺭﺑﺔ diseases affecting humans – Why? Which disease has the most drug candidates ? ﻣﺮﺷﺤﻴﻦ Why does that disease have more drug candidates than hepatitis C? 1. This reflects the current needs of humans- we have too many diseases and currently too few drugs to target them. 2. By far, cancer has the most drug candidates than any other disease. This disease has many more drug candidates than hepatitis C because hepatitis C affects less people worldwide than different kinds of cancer. Cancer, of course, can affect many different organs. On the other hand, hepatitis C only affects the liver.

• Use genetically modified cultured cells to make protein ؟

• Products of Modern Biotechnology – Example of proteins created by gene cloning called recombinant proteins

Types of Biotechnology • • Microbial Biotechnology Agricultural Biotechnology Animal Biotechnology Forensic Biotechnology Bioremediation Aquatic Biotechnology Medical Biotechnology Regulatory Biotechnology

Types of Biotechnology • Microbial Biotechnology – manipulation ﻣﻌﺎﻟﺠﺔ of microorganisms such as yeast and bacteria – Create better enzymes – More efficient decontamination processes for industrial waste product removal – Used to clone and produce large amounts of important proteins used in human medicine

Types of Biotechnology • Agricultural Biotechnology – United Nations Food and Agricultural Org. predicts by 2050, we will need to feed a world population of 9. 1 billion! This requires raising food production by approximately 70%! – brainstorm : gave a few solutions to better feed the world by 2050? 1. use of recombinant technology to genetically modify foods. For example, scientists can create ﺍﻳﺠﺎﺩ better crops that are perhaps more drought ﺟﻔﺎﻑ resistant. 2. They can create ways ﺧﻠﻖ ﻃﺮﻕ to better preserve ﺣﻔﻆ fruits and vegetables so they don't quickly rot ﺗﺘﻌﻔﻦ and get wasted ﺗﺘﻠﻒ. 3. Perhaps create crops that are foul tasting ﻃﻌﻢ ﻏﻴﺮ ﻣﺴﺘﺴﺎﻍ to pests ﺍﻻﻓﺎﺕ including bugs and animals so that the crops survive. 4. Perhaps create ways to have crops grow even in the most extreme conditions. 5. obtain proteins from other novel food sources.

Types of Biotechnology • Agricultural Biotechnology – Plants more environmentally friendly that yield more per acre ( ﺩﻭﻧﻢ genetically engineered) – Resistance to diseases and insects – Foods with higher protein or vitamin content – Drugs developed and grown as plant products – These better plants ultimately reduce production costs to help feed the growing world population

Types of Biotechnology • Agricultural Biotechnology – discuss how you can use this technology in a third world country to create a better corn crop (main crop in that country) that contains all of the 22 essential amino acids ؟

Types of Biotechnology • Will improved crops ﺍﻟﻤﺤﺎﺻﻴﻞ ﺍﻟﻤﺤﺴﻨﺔ that are created to satisfy ﻟﺘﻠﺒﻴﺔ world hunger ﺍﻟﺠﻮﻉ reduce available ﻣﺘﺎﺣﺔ land for biofuel ﻭﻗﻮﺩ ﺣﻴﻮﻱ crops? Discuss This is an ethical question and should lead to some powerful discussions. discuss what is meant by biofuel crops

Types of Biotechnology • Animal Biotechnology – Animals as a source of medically valuable proteins • Antibodies • Transgenic animals ﻣﻌﺪﻟﺔ ﻭﺭﺍﺛﻴﺎ – Animals as important models in basic research • Gene "knockout" experiments ﺗﻀﺮﻳﺐ • Design and testing of drugs and genetic therapies – Animal cloning • Source of transplant organs ﺯﺭﺍﻋﺔ ﺃﻌﻀﺎﺀ

Types of Biotechnology • Animal Biotechnology – transgenic animal: way to achieve ﻟﺘﺤﻘﻴﻖ large scale production of therapeutic proteins from animals for use in humans – Female transgenic animals express therapeutic proteins in milk (contains genes from another source) – Example: human genes coding for clotting ﺗﺨﺜﺮ proteins can be introduced into female goats for production of these proteins in their milk

Types of Biotechnology • Animal Biotechnology – Gene knockout: (gene knock-in) • Disrupt ﺗﻌﻄﻴﻞ a gene in the animal and then look at what functions are affected in the animal as a result of the loss of the gene • This allows ﻳﺴﻤﺢ researchers to determine the role and function of the gene • Since humans are similar to rats and mice, gene knockout studies in rats and mice can lead to better understanding of gene function in humans. • give an example of a gene you would like to knockout in mice? Perhaps, you want to study the role of insulin in preventing ﻣﻨﻊ diabetes. By knocking out the gene coding for insulin, you can observe ﺭﺻﺪ the animal's response (development of diabetes). Then scientists can inject recombinant insulin to the animal and again, study their responses to the treatment

• Forensic Biotechnology( ﺗﻘﻨﻴﺎﺕ ﺍﻟﻄﺐ ﺍﻟﺸﺮﻋﻲ )ﺍﻟﺠﻨﺎﺋﻲ – DNA fingerprinting • Inclusion or exclusion of a person from suspicion ﺍﻟﺸﻚ Paternity cases ﺍﻻﺑﻮﺓ • Identification of human remains ﺗﺤﺪﻳﺪ ﺍﻟﺮﻓﺎﺓ • Endangered species ﺍﻻﻧﻮﺍﻉ ﺍﻟﻤﻬﺪﺩﺓ ﺑﺎﻻﻧﻘﺮﺍﺽ • Tracking and confirmation of the spread of disease ﺗﺘﺒﻊ ﺍﻻﻣﺮﺍﺽ

Types of Biotechnology • Forensic Biotechnology • Based on DNA results from this gel, did the defendant ﺍﻟﻤﺪﻋﻰ ﻋﻠﻴﻪ commit ﺍﺭﺗﻜﺐ this crime ? ﺟﺮﻳﻤﺔ Explain based on the gel results. Based on the gel results, there is evidence that the defendant committed the crime. The position of the bands as well as number of bands on the gel match with the victim's ﺿﺤﻴﺔ blood. It is important to note, that though the bands are very light for the sample from the jeans, they also match the victim's blood.

he figure below represents the results of a DNA gel electrophoresis of DNA collected at a crime scene, a sample from the victim, and samples from suspects. Use the figure to determine which, if any, suspects' DNA matches the crime scene sample. Explain your reasoning.

Example : Compare the DNA collected from the crime scene to determine which of the three suspects was present (click picture to solve)

Example: Paternal Testing Compare the profiles of three men with that of a mother and child to determine the biological father (click picture to solve)

Example: Paternal Testing

Types of Biotechnology • Bioremediation ﺍﻟﻜﻨﺲ ﺍﻟﺤﻴﻮﻱ – The use of biotechnology to process and degrade a variety of natural and man-made substances ﺍﻟﻤﻮﺍﺩ ﺍﻻﺻﻄﻨﺎﻋﻴﺔ • Particularly those that contribute ﺗﺴﺎﻫﻢ to environmental pollution ﺍﻟﺘﻠﻮﺙ ﺍﻟﺒﻴﺌﻲ – Example – stimulated ﺗﺤﻔﻴﺰ growth of bacteria that degrade components in crude oil • 1989 Exxon Valdez oil spill ﺗﺴﺮﺏ in Alaska • 2010 Deep Water Horizon spill promoted research into natural oil-degrading organisms and enzymes

Types of Biotechnology • Bioremediation – adding nutrients to stimulate growth of bacteria to clean up oil spill

Q\ Describe how bacteria are used in pollution reduction operations (oil-eating bacteria) in the oceans?

Types of Biotechnology • Aquatic Biotechnology • Why create ﻳﺨﻠﻖ transgenic salmon overproducing growth hormone? transgenic normal Two different salmon • How does this modified salmon help humans? Answer 1. By creating transgenic salmon that overproduce Growth Hormone, it allows the salmon to have fast growth rates over a short period of time. Answer 2. This modified salmon helps humans because it decreases the time and expenses required to grow the salmon for market sale.

Types of Biotechnology • Medical Biotechnology 1. Involved with the whole spectrum ﻛﻞ ﻃﻴﻒ of human medicine • Preventive medicine ﺍﺩﻭﻳﺔ ﻭﻗﺎﺋﻴﺔ • Diagnosis of health and illness ﺗﺸﺨﻴﺺ ﺍﻟﺼﺤﺔ ﻭﺍﻟﻤﺮﺽ • Treatment of human diseases ﻋﻼﺝ ﺍﻻﻣﺮﺍﺽ ﺍﻟﺒﺸﺮﻳﺔ 2. New information from Human Genome Project • Gene therapy ﺍﻟﻌﻼﺝ ﺍﻟﺠﻴﻨﻲ 3. Stem cell ﺍﻟﺨﻼﻳﺎ ﺍﻟﺠﺬﻋﻴﺔ technologies

• Stem cells – grown in lab and then treated with different chemicals to allow ﻳﺴﻤﺢ them to develop into specific kinds of tissues needed for transplant ﺍﻷﻌﻀﺎﺀ ﺯﺭﺍﻋﺔ • Current use ﺍﻟﺤﺎﻟﻲ ﺍﻻﺳﺘﻌﻤﺎﻝ : stem cells are used for diabetes; spinal cord injuries ﺍﻟﺸﻮﻛﻲ ﺍﻟﺤﺒﻞ ﺍﺻﺎﺑﺎﺕ • Can you gave other diseases that scientists are testing with stem cells? • • • replace neurons damaged by spinal cord injury, stroke, Alzheimer’s disease, Parkinson’s disease or other neurological problems; produce insulin that could treat people with diabetes and heart muscle cells that could repair damage after a heart attack; or replace virtually any tissue or organ that is injured or diseased Remember, most of these diseases are only be tested with stem cell therapy in animal models. They are not being used in the clinic ﺍﻟﻌﻴﺎﺩﺍﺕ yet!!!

Q\explain why scientists are doing more research using embryonic vs. adult stem cells.

• Medical biotechnology – Genes are headline ﻋﻨﻮﺍﻥ ﺭﺋﻴﺴﻲ news items

• How will medical biotechnology change our lives in the years ahead ? ﻗﺎﺩﻣﺔ – Human Genome Project ﻣﺸﺮﻭﻉ ﺍﻟﺠﻴﻨﻮﻡ ﺍﻟﺒﺸﺮﻱ • Research on the function of human genes and controlling factors that regulate genes – Human proteome ﺑﺮﻭﺗﻴﻨﻮﻡ ﺍﻟﺒﺸﺮ • Group of proteins responsible for activity in a human cell

• How will medical biotechnology change our lives in the years ahead? – Single Nucleotide Polymorphisms (SNPs) • Single nucleotide changes (ﺗﺘﻐﻴﺮ mutations) in DNA sequences that vary ﺗﺨﺘﻠﻒ from individual ﻓﺮﺩ to individual (SNP : is a variation in a single nucleotide that occurs at a specific position in the genome) • These variations ﺍﻻﺧﺘﻼﻓﺎﺕ are the cause ﺳﺒﺐ of some genetic diseases ﺍﻻﻣﺮﺍﺽ ﺍﻟﻮﺭﺍﺛﻴﺔ (sickle cell anemia )ﻓﻘﺮ ﺍﻟﺪﻡ ﺍﻟﻤﻨﺠﻠﻲ • SNPs will help identify genes involved ﻣﺸﺎﺭﻛﺔ in medical conditions ﺣﺎﻻﺕ ﻣﺮﺿﻲ including arthritis ﺍﻟﺘﻬﺎﺏ ﺍﻟﻤﻔﺎﺻﻞ , stroke ﺳﻜﺘﻪ ﺩﻣﺎﻏﻴﺔ , cancer, heart disease, diabetes, and behavioral ﺍﻟﺴﻠﻮﻛﻴﺔ and emotional ﺍﻟﻨﻔﺴﻴﺔ illnesses

• Example of SNPs and breast cancer • Identification of SNPs in BRCA 1 and BRCA 2 genes involved ﻣﺸﺎﺭﻛﺔ in promoting ﺗﻌﺰﻳﺰ breast cancer led ﻗﺎﺩ to development ﺗﻄﻮﻳﺮ of better targeted treatments for people who have those specific gene mutations ﻃﻔﺮﺍﺕ ﻣﺤﺪﺩﺓ ﺟﻴﻨﻴﺔ • Can you think of how this knowledge might be useful for someone who is not already diagnosed with cancer? (think of your basic knowledge of genetics. ) Assume ﺍﻓﺘﺮﺽ that friends mom and sister both had breast cancer and found out that they both had the BRCA 1 gene mutation. His doctor would counsel he to be tested for this mutation too. Then, if he has the mutation, the doctor could monitor he more closely for developing breast cancer. If he was to develop this cancer, they would be able to catch it early on when he has a better chance of being treated successfully. It is important to note that even if he has tested and have this genetic mutation, it does NOT mean that he will definitely develop cancer. It just means that he has an increased risk! It is also important to note that both men and women can develop breast cancer

• Example of how we can benefit ﻧﺴﺘﻔﺎﺩ from the human genome project • Based on the figure ﺍﻋﺘﻤﺎﺩﺍ ﻋﻠﻰ ﺍﻟﺸﻜﻞ , why doesn't person 2 develop a genetic disease due to the SNP (G → T)? Since there can be multiple codons coding for the same amino acid, then even if person 2 has a SNP, the new codon can still code for the same amino acid as in person 1. Thus, the same protein will ultimately be made and it will therefore have the correct function. This type of mutation is called silent since the nucleotide change did not affect a change in the amino acid sequence. Person 3 had a SNP that changed the codon so it coded for a different amino acid. Thus, this will affect the way the protein is folded ﺍﻟﺘﻔﺎﻑ and, if not in the proper structure ﺍﻟﺼﺤﻴﺢ ﺍﻟﺘﺮﻛﻴﺐ , it will have impaired ﺿﻌﻒ function or perhaps no function.

• Gene therapy technology – Replacing ﺍﺳﺘﺒﺪﺍﻝ or augmenting ﺍﺿﺎﻓﺔ defective genes ﺟﻴﻦ ﻣﻌﻴﻦ with normal copies of the gene Still have barriers ﺗﺤﺪﻳﺎﺕ to overcome before this technology becomes safe and effective • Obstacles include ﺗﺸﻤﻞ ﺍﻟﻌﻘﺒﺎﺕ : – How can normal genes be delivered to all cells in the body? – What are the long-term effects ﺍﻻﺛﺎﺭ ﻋﻠﻰ ﺍﻟﻤﺪﻯ ﺍﻟﻄﻮﻳﻞ of introducing ﺍﺩﺧﺎﻝ extra ﺟﻴﻨﺎﺕ ﺇﺿﺎﻓﻴﺔ genes in humans? – What must be done to ensure ﺿﻤﺎﻥ the proper protein is made after the genes are delivered to the body?

• How will medical biotechnology change our lives in the years ahead? – Regenerative medicine ﺍﻟﻄﺐ ﺍﻟﺘﺠﺪﻳﺪﻱ • Genetically modifying stem cells of patients to treat genetic disease • In future scientists will be able to… 1. Isolate adult stem cells from a patient with a genetic disorder ﺍﺿﻄﺮﺍﺑﺎﺕ ﻭﺭﺍﺛﻴﺔ 2. Genetically manipulate ﺗﻼﻋﺐ these cells by gene therapy 3. Reinsert ﻋﺎﺩﺓ ﺍﺩﺧﺎﻝ the cells into the same patient to help and treat their genetic disease

Insulin is one of the important pharmaceutical products produced commercially by genetically engineered bacteria. Before this development, commercial insulin was isolated from animal pancreatic tissue. Microbial insulin has been available since 1982. The human insulin gene is introduced into a bacterium like E. coli. Two of the major advantages of insulin production by microorganisms are that the resultant insulin is chemically identical to human insulin, and it can be produced in unlimited quantities.

What is genetic engineering and what is it used for? The ability to manipulate and analyse DNA using genetic engineering techniques (recombinant DNA technology) was foreseen in the mid 1960 s and came to fruition in the early 1970 s. The technology, The basic tools of genetic engineering The techniques for isolating, cutting and joining molecules of DNA, developed in the early 1970 s, have provided the foundations of our current technology for engineering and analysing nucleic acids. These allow fragments of DNA from virtually any organism to be cloned in a bacterium by inserting them into a vector (carrying) molecule that is stably maintained in the bacterial host.

Isolation and purification of nucleic acids Biochemical techniques for preparing large quantities of relatively pure nucleic acids from microbial cells are an essential pre-requisite for in-vitro gene technology. The first step in the isolation of nucleic acids is the mechanical or enzymatic disruption of the cell to release the intracellular components that include the nucleic acids. Once released from the cell, the nucleic acids must be purified from other cellular components such as proteins and polysaccharides to provide a substrate of appropriate purity for nucleic acid modifying enzymes. The released nucleic acids are recovered using a combination of techniques including centrifugation, electrophoresis, adsorption to inert insoluble substrates or precipitation with non-aqueous solvents

Cutting DNA molecules The ability to cut molecules of DNA, either randomly or at specific target sites, is a requirement for many recombinant DNA techniques. DNA may be cleaved using mechanical or enzymatic methods. Mechanical shearing is non-specific and results in the production of random DNA fragments, which are often used to generate genomic libraries (see Section 4. 4. 5). When DNA molecules are mechanically sheared it is not possible to isolate a specific fragment containing, for example, a particular gene or operon. In contrast, when the DNA is cut using restriction endonucleases, which recognize and cleave specific target base sequences in double-stranded (ds)DNA, specific fragments can be isolated. Restriction endonucleases cut the phosphodiester backbone of both strands of the DNA to generate 3 OH and 5 PO 4 termini. Several hundred restriction endonucleases have been isolated from a wide variety of microbial species. Restriction endonucleases are classified into groups with distinct biochemical properties; type II enzymes are the main class used for genetic engineering purposes

Restriction endonucleases are named according to the species from which they were originally isolated; enzymes isolated from Haemophilus influenzae are designated Hin, those from Bacillus amyloliquefaciens, Bam, etc. When more than one type of enzyme is isolated from a particular strain or species, the strain and isolation number (in roman numerals) are added to the name. Thus the three restriction endonucleases isolated from H. influenzae strain Rd are designated Hind. I, Hind. II and Hind. III.