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by: Robert Notetaker
Robert Notetaker

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Genetic Engineering, Modification and use of stem cells in gene therapy.
DNA: Identify and Disease
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This 7 page Class Notes was uploaded by Robert Notetaker on Monday March 21, 2016. The Class Notes belongs to at Brigham Young University - Idaho taught by in Winter 2016. Since its upload, it has received 12 views. For similar materials see DNA: Identify and Disease in Biology at Brigham Young University - Idaho.


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Date Created: 03/21/16
Gene Therapy: A treatment for genetic Disease CFTR Gene TranscribedDFTR mRNA translated CFTR Protein Does not work. CFTR helps with salt and H20 balance. Gene creates a CFTR Protein that does not work. Cut out the gene with molecular scissors and cut and paste in plasmid vector and ligate. Needs a human promotor. We are not making a transgenic organism. We will then put this into a virus. This virus will inject the working copy of the CFTR plasmid. Stable integration. Normal CFTR gene gets intercepted into a chromosome. Gene Therapy:“Vectors” Suitcase. Most common vector for gene therapy: Virus Virus-Immune Response. Efficiency is high. Liposomes (balls of membrane that hold DNA.) Plasmid DNA. They then will fuse and then go into the cell. Still can have an immune response sometimes. Low immune response. Naked DNA- Just injecting DNA without any suitcase. Just the DNA. Low immune response. The efficiency is very low. What makes a good candidate for gene therapy? Affected tissue is easily accessible. Only one gene. We need a good understanding biology of the disease. We cannot cure a disease without understanding it. Mutated gene does not interphase with the cells function. Clumps are BAD!!! Just replacing the gene is not enough. Crisper: Allows to cut specific genes. Cuts specific genes. You have to know where the gene is. Ex-Vivo viruses never enters the body, never ever sees the virus. More efficient. We only treat certain cells. In Vivo – Viruses enter the body. Less efficient. The viruses infect other cells… How do we know gene therapy works: It works, you do not get sick and die. The viruses could break a gene which is very bad. Liposome-wont integrate Naked DNA: No Size Limit, no immune response Stem Cells Powers Ability to differentiate which is called potency We have different levels of potency Mullipotent-Already differentiated (Not stem cells, is a skin cell) Unipotent-Only one kind-skin stem cell. Multipotent-One of a few kinds of cells (homatopoetic Cells) Totipotent: Any cell and placental tissue. Also called Morula Cells Pluripotent:Can become any cell in the body.Also called embryonic stem cells Genetic Modification Artificial Selection (Breeding) Old 1,000’s of years Produce Desirable traits More Natural Downside: Very Slow(Years), Randomness(Less control over outcome), Mutagenic Breeding Old 150 Years Chemicals for mutagenic ray(radiation, uv) (ENU-breaks DNA), Fast 1-2 generations, What genes are mutated. Ethical Consideration Genetic Engineering Transgenic, Lab-based, not random, very precise, foreign genes, Apricot and Plum, Antifreeze ti Plasmid antibiotic resistance gene on it, also has a origin of replication and tomato promoter. 1. Agrobacteria 2. Gene Gun A mode of delivery. (Envelopes) Agrobacteria Tumefaciens – infects plant cells and inserts ti plasmids into plant chromosomes. Ti transfer to agrobacterium and infect plant cells. The agrobacterium gets inside the cell and gets together with the chromosome in the nucleus and cuts out and inserts the promoter and the gene. Toti Potency: Ability of any plant cell to become a whole plant. Gene Gun: Plasmid: Tomato PromoterAntifreezeantibiotic Resistance Animal GMO’s Modes of delivery 1. Microinjection 2. Virus- Take cell of interest and then get a virus and then put gene into virus and then it will inject the gene into the nucleus of the cell. Gene Delivery: How do we inject DNA into animal cells? Microinjection, viruses, and then combination of DNA with stem cells. Robert Macias P R E PAR A T I O N & P O N D E R I N G-4-7 stem cells 1.What are stem cells? They are unspecified cells capable of renewing themselves through cell division. They can become any cell that is found in the body. 2.What are some of the properties of stem cells that make them interesting to scientists? They have the ability to become any cell in the body. They can generate replacements for cells that are lost through normal wear and tear, injury, or disease. They can also be used to treat diabetes and heart disease. They can replicate many times and can yield millions of cells. They can do this for about a year. 3.What does it mean when a cell is referred to as differentiated? What causes a stem cell to become differentiated? This is when the stem cell is when it receives a signal to become a somatic cell in the body. This is the point where it is no longer a stem cell anymore. This is done by chemicals. 4.Why are embryonic stem cells preferred over adult stem cells? Adult stem cells are able to generate cell types of the tissue it resides in. Embryonic stem cells have the potential to become any cell that is found in the body and not just one type of cell from the body. 5.Explain the difference between totipotent, pluripotent, and multipotent. Which types of stems cells is associated with each term? Pluripotent cells can be any type of cell from the body, they can not make extra embyryonic tissues such as amnion, chorion, and other parts from the placenta. Multipotent is the ability to develop into more than one cell type of the body. Totipotent have the ability to be any cell type in the body plus all cell types that can make up extraembryonic tissues such as the placenta. 6.Why is cord blood a valuable resource? Blood is needed for growth. 7.What are the obstacles that must be overcome before the potential uses of stem cells in cell therapy will be realized? The body will naturally reject the injected stem cells from a healthy person’s body. So there is a medication that will help reduce rejection from the body. 8.What is the main ethical issue associated with the use of embryonic stem cells? 9.What are induced pluripotent stem cells (iPSCs)? Embryonic cells are attained by embryos which in turn, many people do not like that. There is a huge debate on whether that is killing a human if these cells are harvested for science. These cells have all the potential to become a cell that it was near. They can change back from a stem cell for that specific cell and back again. They can be used for therapy for other people who are sick or dying to replace their non-functional cells. 10.iPSCs not only help scientists overcome an ethical issue but a technical one as well that has to do with tissue rejection. Explain why iPSCs might be better than ESCs on a technical level. With iPCSs we do not have to worry about even dipping into immoral matters of debate or issue. We have all the resources we need to make new cells for that patient, as long as we have someone willing to help out with the process. 11.How could iPSCs be used to understand disease better? The more we understand about the processes we can see how they work and function and how we can beat aging or even elongate life just a bit longer as we continue to study and know about these cells. We should use viruses, DNA, and stem cells to help us understand everything about disease, death, aging, regenerating, and much more that we can find out. 12.Is there a line that should not be crossed in the use of human gene and stem cell therapy? For example, what are your feelings about modifying germ line tissue (sperm & eggs) so that any changes in the individual would be passed on to their offspring? I think anything helpful to help get rid of disease is fine. But altering ourselves just for fun and selecting choices could make less uniqueness in people over time. Any changes regarding the elimination of disease would be great, but when the times comes for picking what we want for our children, I do not want everyone to look the same with those blue eyes and gold hair or whatever is in. Honestly what happens happens. If it is not supposed to happen then history will take care of things like this. But then again, God did not stop the production of the Atom Bomb and its use, and again, dozens of assassinations targeted towards Hitler never succeeded. If technology can do things that we find interesting and useful then it will be used accordingly as long as morals are not broken… 13.Think of two ways you can make your opinions on these issues known to your local or state government. I could participate in a city council and be involved in these issues that will very much become a reality very soon. I can also contact my local congressman regarding the matter and issues like this can eventually be discussed over time. 14.Do the consensus opinion of American society and the government on gene therapy and stem cell research seem to be in harmony with yours or the majority of those of the LDS faith? Yes, so far. We are not at the testing human trials stage at full blown gene therapy nor helping babies out with their defunct genetic code yet. So far help has been done in science making us live with these issues and someday living free of them. We use viruses to insert genes into the body or make vaccines and use them to help with our health and living conditions. My faith we just have to watch out for giving our sperm or egg to another for an invitro fertilization for someone who cannot have kids. After all, that is part of us that is spiritually and physically connected to ourselves and will confuse genetic tests way down the line in the future… 15.What other questions do you have about stem cells? Can we use them to actually replace the non-functioning cells in our body? Can we replace these cell? Are we far away from this type of therapy, will it even work with the te4chnology we have today?


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