Biotechnology Intro PDF

BIOTECHNOLOGY  Vanessa Jason Biology Roots BIOTECHNOLOGY Vanessa Jason Biology Roots Biotechnology produces many products that you have probably used or consumed- it is a...

BIOTECHNOLOGY  Vanessa Jason Biology Roots BIOTECHNOLOGY Vanessa Jason Biology Roots Biotechnology produces many products that you have probably used or consumed- it is a part of our daily lives. Vanessa Jason Biology Roots Simply put, biotechnology is the use of an organism to achieve a specific goal. It can be as simple as using yeast to bake, or as advanced as growing animal parts to replace diseased human organs. Types of biotechnology- - Simple: growing mold for penicillin (a natural antibiotic) - Advanced: many modern forms of biotechnology involve the manipulation of DNA, such as GMOs or gene therapy. Vanessa Jason Biology Roots © Biology Roots, LLC  REFRESHER DNA 🡪 RNA 🡪 Proteins Genes are segments of DNA that code for a specific protein (such as insulin, enzymes, proteins that are fatal to pests and and can act as natural pesticides, etc.) *All cells within an organism have the same DNA, but not all genes are expressed, depending on the cell’s function For example, insulin is produced in pancreatic cells, but not in heart cells (gene for insulin is expressed in pancreatic cells; repressed in others) * Pathogens as Vectors Viruses and bacteria have naturally occurring enzymes and proteins that can manipulate DNA. Upon discovering this, many of our latest biotechnologies use viruses or bacteria as vectors to modify DNA.  Vanessa Jason Biology Roots Vanessa Jason Biology Roots Vanessa Jason Biology Roots BIOTECHNOLOGY Examples of biotechnology: 1. DNA Fingerprinting (popular in forensics to compare known DNA samples to sample in question using a technique called gel electrophoresis). 2. Genetically modified organisms – organisms that have had their DNA modified to express a desirable trait. Sometimes, GMOs contain DNA from two species (transgenic); this can be achieved using bacteria and/or viruses as vectors to cut and paste DNA). 3. Gene Therapy- replacing a defective gene with a healthy version using viruses as vectors to deliver DNA. Vanessa Jason Biology Roots © Biology Roots, LLC  Vanessa Jason Biology Vanessa Roots Jason Biology Roots TRANSGENIC ORGANISMS Vanessa Jason Biology Roots Transgenic organisms have had their DNA manipulated somehow to express a foreign gene. Trans= transfer; across Genic= gene Transgenic= a transfer of a gene from one species to another. Some transgenic organisms are GMOs (genetically modified organisms). Only GMOs that contain DNA from two species are transgenic. Transgenic goats have been modified to produce spider silk in their milk. Spider silk is a very popular material due to its tensile strength, but harvesting it from spiders proved to be difficult.Vanessa Jason Biology Roots © Biology Roots, LLC COMMONLY USED GMO s Percentage of GMO Crops in the US Corn 94% GMO Contain a gene from bacteria that is Cotton 94% GMO resistant to insects (this particular gene produces a protein that kills the larvae of certain pests.) Sugar 99.9% GMO beets Contain a gene from bacteria that is resistant to herbicides. Soybeans 94% GMO The top 5 GMO crops in Canada include canola, corn, potatoes, soybeans, sugar beets and alfalfa. These GMOs are transgenic because they contain DNA from other species (bacteria.)  Vanessa Jason Biology Vanessa Roots Jason Biology Roots TRANSGENIC ORGANISMS Vanessa Jason Biology Roots HOW are transgenic organisms created? With the help of bacteria in a process known as recombinant DNA technology: the joining of DNA from two different species. In order to insert a gene from one species into an other species, we use bacteria and enzymes in plasmid based transformation. Recombinant DNA= molecules of DNA from two different species that are inserted into a host organism to produce new genetic combinations Vanessa Jason Biology Roots https://www.youtube.com/watch?v=FAMRQz7fOaE © Biology Roots, LLC  Vanessa Jason Biology Vanessa Roots Jason Biology Roots Vanessa Jason Biology Roots PLASMID TRANSFORMATION- USES DNA Recombinant Technology (or DNA cloning) is the process of making many copies of a specific DNA sequence in order to make lots of protein. The copies are typically made in bacteria by introducing foreign DNA into a plasmid (transformation.) Examples include: -Production of insulin for diabetics -tPA (life saving protein that can dissolve blood clots) Vanessa Jason Biology Roots © Biology Roots, LLC  Vanessa Jason Biology Vanessa Roots Jason Biology Roots Vanessa Jason Biology Roots PLASMID TRANSFORMATION To “cut and paste” DNA from one organism into another, enzymes are required. -Restriction enzymes- enzymes found in bacteria that recognize and bind to specific sequences of DNA, called restriction sites, and cut the DNA at these specific base pairs. -DNA Ligase- an enzyme that seals (or pastes) DNA nucleotides together (also used in DNA replication). Vanessa Jason Biology Roots © Biology Roots, LLC  Vanessa Jason Biology Vanessa Roots Jason Biology Roots Vanessa Jason Biology Roots PLASMID TRANSFORMATION Bacteria can take up foreign DNA in a process known as transformation. Transformation is used to target a gene of interest from one organism and introduce it into the genome of bacteria to create recombinant organisms. Plasmid transformation uses circular rings of DNA from bacteria known as plasmids. The bacteria can replicate quickly, and create many copies of DNA, and therefore many copies of a protein. Vanessa Jason Biology Roots © Biology Roots, LLC  Vanessa Jason BiologyJason Vanessa RootsBiology Roots For example, insulin can be produced in mass quantities to treat diabetes. Insulin is a Vanessa Jason Biology Roots protein produced in the pancreas. It’s primary function is to regulate blood sugar. Human pancreas cell Bacteria cell with recombinant DNA DNA (includes gene for insulin) Recombinant DNA produces insulin Plasmid (cloning Human DNA Restriction enzymes DNA ligase is used to Insulin can be vector) containing gene are used to cut the seal the bonds at the harvested from that codes for DNA so that only the sticky ends and produce bacteria and insulin gene of interest is recombinant DNA (and prescribed to Vanessa Jason Biology Roots present transgenic bacterium) diabetics. © Biology Roots, LLC  Vanessa Jason Biology Vanessa Roots Jason Biology Roots Vanessa Jason Biology Roots PLASMID TRANSFORMATION The specific plasmids used for recombinant DNA technology carry a gene for antibiotic resistance. Not all bacteria can be used for plasmid transformation. Selection- to select for the bacteria that can successfully produce copies of the gene/protein, bacteria are grown on an antibiotic plate. The ones that survive have the recombinant plasmid. Further testing is required to make sure the foreign DNA was successfully integrated into the plasmid. Vanessa Jason Biology Roots © Biology Roots, LLC  Vanessa Jason Biology Vanessa Roots Jason Biology Roots Vanessa Jason Biology Roots GEL ELECTROPHORESIS Gel electrophoresis is a laboratory technique used to distinguish DNA fragments of different lengths. In gel electrophoresis, DNA is separated and pushed by an electrical field through an agarose gel. “Electro” “Phoresis” electricity to carry *The same principles of gel electrophoresis can be used for other biomolecules, such as RNA and proteins. Vanessa Jason Biology Roots © Biology Roots, LLC  Vanessa Jason Biology Roots GEL ELECTROPHORESIS Vanessa Jason Biology Roots Because DNA molecules are negatively charged (-), they can travel across a medium (gel) carried by electrical currents. The larger molecules are not pushed as far as the smaller ones. Vanessa Jason Biology Roots Which direction do you think the DNA travels and why? Vanessa Jason Biology Roots © Biology Roots, LLC  Vanessa Jason Biology Vanessa Roots Jason Biology Roots Vanessa Jason Biology Roots https://www.youtube.com/watch?v=9DDF5DKr82g Vanessa Jason Biology Roots https://www.youtube.com/watch?v=ZG7KtuBfReo © Biology Roots, LLC  Vanessa Jason Biology Roots GEL ELECTROPHORESIS Vanessa Jason Biology Roots Potential fathers cub Vanessa Jason Biology Roots Reading a gel- the gel is stained with a special dye that can be seen under UV light. The picture at right is a hypothetical gel used for a bear cub paternity test. M= mother; 1, 2, 3 are potential fathers. Ideally the bands formed from the offspring’s DNA would depict a combination of both parents. Based on the gel electrophoresis results, which male do you think the Vanessa Jason Biology Roots father of the cub is? © Biology Roots, LLC  Vanessa Jason Biology Roots GEL ELECTROPHORESIS Vanessa Jason Biology Roots Samples are compared to a DNA marker (or “size standard”) that contains known sizes of DNA fragments in base pairs. Vanessa Jason Biology Roots The DNA marker is shown at ~6000 bp the left of the gel. The unknown sample is on the right. We can determine the base pairs of the ~3500 bp unknown sample by comparing it the fragments to the DNA ~1500 bp marker. Vanessa Jason Biology Roots © Biology Roots, LLC  GEL ELECTROPHORESIS Vanessa Jason Biology Roots Vanessa Jason Biology Roots Why use gel electrophoresis? Forensics- Gel electrophoresis can be used as a means of DNA fingerprinting. Bands of DNA in gel electrophoresis can have patterns specific to an individual person, which comes from the size and sequence of the DNA in question. Suspects can be eliminated if their DNA pattern does not match the pattern of DNA molecules found at the crime scene. Or, on the other hand, pose as evidence that an individual is a suspect. Zoology- mapping endangered species, exploring pedigrees Other uses- to analyze PCR (polymerase chain reaction) results (think COVID tests); to isolate and analyze genes associated with a particular illness Vanessa Jason Biology Roots © Biology Roots, LLC  Vanessa Jason Biology Vanessa Roots Jason Biology Roots Vanessa Jason Biology Roots GEL ELECTROPHORESIS Applications of Gel Electrophoresis Vanessa Jason Biology Roots https://www.youtube.com/watch?v=WPTWOGm3iN0 © Biology Roots, LLC  Vanessa Jason Biology Vanessa Roots Jason Biology Roots Vanessa Jason Biology Roots PCR Polymerase Chain Reaction- lab technique that is used to make lots of copies of DNA. https://www.youtube.com/watch?v=hO3mTqrEeq8 How does PCR relate to other biotechnology techniques we’ve Vanessa Jason Biology Roots learned about so far? © Biology Roots, LLC  Vanessa Jason Biology Vanessa Roots Jason Biology Roots Vanessa Jason Biology Roots PCR PCR has many applications, it can be used in all fields of science, including biotechnology Examples: -To amplify copies of DNA that was found at a crime scene. -To make copies of DNA during amniocentesis (to test for chromosomal abnormalities) -To amplify copies for the sensitive detection of pathogens (Covid PCR test) Ultimately, the more copies of a particular DNA segment you have, the more accurate detection (or test results) will be. Vanessa Jason Biology Roots © Biology Roots, LLC  Vanessa Jason Biology Vanessa Roots Jason Biology Roots HUMAN GENOME PROJECT Vanessa Jason Biology Roots Genome= all the genes found in an organism Why would be want to understand the human genome? We can target which genes, or their mutations, are associated with certain genetic diseases, such as cystic fibrosis or cancers that run in the family. Improved diagnosis of genetic diseases Earlier detection of genetic predispositions to disease We can predict how individuals might respond to certain medicinal drugs. Gene therapy Human Genome Project (HGP) was an international research project initiated in 1990. Nearly the entire human genome was mapped to recognize and understand our genes. Vanessa Jason Biology Roots © Biology Roots, LLC  Vanessa Jason Biology Vanessa Roots Jason Biology Roots Vanessa Jason Biology Roots HUMAN GENOME PROJECT The technology and principles behind the human genome project has expanded to the following applications: Energy sources and environmental applications Risk assessment of certain diseases Bioarcheology, anthropology, evolution, and human migration DNA forensics (identification) Agriculture, livestock breeding, and bioprocessing Vanessa Jason Biology Roots © Biology Roots, LLC  Vanessa Jason Biology Vanessa Roots Jason Biology Roots Vanessa Jason Biology Roots GENE THERAPY Gene Therapy is a process to replace defective genes with healthy ones to treat genetic diseases. A popular gene therapy process borrows “technology” from viruses. When viruses infect our cells, they essentially hijack our cells and use our cellular mechanisms to create more of their own genetic material. We can alter viruses to be nonvirulent, and deliver healthy copies of genes into our cells. Gene therapy typically uses retroviruses or adenoviruses to deliver genes, in what are known as “viral vectors”. Vanessa Jason Biology Roots © Biology Roots, LLC  Vanessa Jason Biology Vanessa Roots Jason Biology Roots Vanessa Jason Biology Roots GENE THERAPY Retroviruses are a popular choice for aiding gene therapy. Retroviruses have an RNA genome. When they infect a host cell, they synthesize complementary DNA from their RNA (this is a unique process called reverse transcription.) The DNA copy is inserted into the host cell. The host cell then starts making viral proteins. Gene therapy- In order to use retroviruses in gene therapy, we transcribe a version of healthy DNA for a particular gene into RNA. The retrovirus will make a copy of DNA from the RNA with the healthy allele, and insert it into the host cell. From there, the host cell starts making the proteins that they need. Vanessa Jason Biology Roots © Biology Roots, LLC  Vanessa Jason Biology Vanessa Roots Jason Biology Roots Vanessa Jason Biology Roots GENE THERAPY There are two general types of gene therapy: Gene Therapy In Vivo Ex Vivo Modified DNA or viruses Cells and tissue are taken are injected directly INTO OUT of the patient, the patient’s tissues. modified outside the body, then injected back into the tissues. Vanessa Jason Biology Roots © Biology Roots, LLC  Vanessa Jason Biology Vanessa Roots Jason Biology Roots Vanessa Jason Biology Roots GENE THERAPY https://www.youtube.com/embed/19fSG4Skvws?end=272 Vanessa Jason Biology Roots © Biology Roots, LLC  Vanessa Jason Biology Vanessa Roots Jason Biology Roots CRISPR Vanessa Jason Biology Roots CRISPR is a natural defense mechanism found in bacterial cells against viruses. “Clustered Regularly Interspaced Short Palindrome Repeats”- the term used to describe a repeating palindrome pattern found in E. Coli that gained the attention of scientists in the 1980s! CRISPR is found between spacer DNA, and follows a palindrome pattern (for example): Vanessa Jason Biology Roots © Biology Roots, LLC  Vanessa Jason Biology Vanessa Roots Jason Biology Roots CRISPR Vanessa Jason Biology Roots Upon studying these palindrome DNA sequences, scientists found they matched perfectly with viral DNA, particularly bacteriophages, which are viruses that invade bacteria. “CRISPR-Cas” provides sequence-specific adaptive immunity in bacteria by snipping viral DNA and integrating short virus DNA sequences in the bacteria's CRISPR locus of its genome, allowing the cell to recognize and clear viral infections. ”Cas” refers to a special enzyme that is required for this specific type of immunity. Essentially, the CRISPR system can cut and add DNA. This made scientists think, what if we could cure genetic diseases by cutting out mutations and adding healthy DNA sequences? Vanessa Jason Biology Roots © Biology Roots, LLC  Vanessa Jason Biology Vanessa Roots Jason Biology Roots CRISPR Vanessa Jason Biology Roots The CRISPR system has been modified to work with eukaryotic DNA, such as animals. CRISPR has 2 main components: RNA, and a protein. The protein used in the lab is called Cas9, so sometimes you see it written as CRISPR-Cas9 https://www.youtube.com/watch?v=9q8AGst7KiY Vanessa Jason Biology Roots © Biology Roots, LLC  Ethical Concerns Vanessa Jason Biology Vanessa Roots Jason Biology Roots Vanessa Jason Biology Roots Biotechnology has the potential to provide benefits to people and societies, but it can also have unintended consequences. It is important that biotechnology innovations be carefully tested and analyzed before they are released for general use. Clinical trials and government regulation help ensure that biotechnology products placed on the market are safe and effective. However, sometimes new information becomes available that makes companies and government agencies reconsider the safety or utility of an innovation. (For example when medicine is recalled). In addition, biotechnology innovations may raise new ethical questions about how information, techniques, and knowledge should or shouldn’t be used. Some of these relate to privacy and non-discrimination. For instance should your health insurance company be able to charge you more if you have a gene variant that makes you likely to develop a disease? How would you feel if your school or employer had access to your genome? Other questions relate to the safety, health effects, or ecological impacts of biotechnologies. For example, crops genetically engineered to make their own insecticide reduce the need for chemical spraying, but also raise concerns about plants escaping into the wild or interbreeding with native populations (potentially causing unintended ecological consequences). Biotechnology may provide knowledge that creates hard dilemmas for individuals. For example, a couple may learn via prenatal testing that their baby has a genetic disorder. Similarly, a person who has her genome sequenced for the sake of curiosity may learn that she is going to develop an incurable, late-onset genetic disease, such as Huntington's. Scientific research and development can make new information, techniques, and knowledge available. However, science alone cannot answer questions about how these techniques should or shouldn’t be used. It's important for all members of society to have their voices heard in the conversation about biotechnology inventions and products that can affect our everyday lives. Vanessa Jason Biology Roots © Biology Roots, LLC