Improved sanitation systems Surgery with anesthesia Vaccines and antibiotics And the fourth will be Gene Therapy

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Slide 2 Slide 3 Improved sanitation systems Surgery with anesthesia Vaccines and antibiotics And the fourth will be Gene Therapy Slide 4 The selective delivery of genes into a patients cells create missing proteins replace defective disease causing genes Over 4,000 conditions are caused by damage to a single gene, many others by several genes Slide 5 Ex-vivo outside the body adding blood cells with new DNA to the blood stream but has a limited time span Using bacteria like E.coli to produce billions of copies of a human gene Using viruses to carry the gene to specific cells in the body Slide 6 A cutting enzyme called aRestriction enzyme It cuts foreign DNA that enter bacteria, e.g. Eco. R1 A circular DNA molecule from a bacteria called aPlasmid The E. coli bacteria has dozens of different plasmids A section of the human DNA containing the gene you need An enzyme to join the bacterial DNA and human DNA calledligase Slide 7 Found in many bacteria A defense mechanism which cuts foreign DNA The DNA is not cut randomly, but at specific sequences called Recognition sequences The restriction enzyme Eco.R1 found in strain C, E. coli bacteria They dont make straight cuts, but produce sticky ends These sticky ends can rejoin by forming hydrogen bonds and the sugar-phosphates rejoining with the help of the enzyme ligase The DNA produced by restriction enzymes cutting is called gDNA We know of 200 different restriction enzymes Slide 8 The Plasmid pSC101 was isolated from E. coli. It is useful for gene therapy because it has only one sequence of GATTC in its entire molecule Plasmids can be used to replicate DNA segments of up to 4,000 base pairs in length For longer length viruses must be used. Slide 9 Using Bacterial Plasmid Cut the Bacterial Plasmid using restriction enzyme called Eco R1 Slide 10 Slide 11 This produces a hole in the circular Plasmid DNA Eco. R1 only cuts at the sequence GAATTC This produces two identical ends Slide 12 Also treat human DNA with Eco.R1 and you get a section of DNA containing the gene you want with two complementary ends as well! Short sequence of DNA with sticky ends approaches a Plasmid. Human DNA Plasmid Slide 13 Slide 14 Bacterial DNA Human DNA The newly completed piece of DNA is called Recombinant DNA Slide 15 Gene cloning can now be used to produce millions of copies Slide 16 A Plasmid with human DNA is put back into a E. coli The bacteria expresses the DNA. The bacteria then divides to produce millions of copies bacteria and human gene. Slide 17 A Plasmid cut by a restriction enzyme Segment of DNA approaches Foreign DNA spliced into Plasmid by DNA ligase joins phosphates and sugars Plasmid enters bacterial cell Slide 18 Viruses can be used to transfer large pieces of human DNA to a location in the Human body. Viruses are made up of a molecule of nucleic acid and a protein coat. Their nucleic acid codes for protein coats, replication, enzymes to break-in and out of particular cells. You remove the part of the nucleic acid that does not code for the above functions and add foreign DNA. The virus can then be put into the blood stream and it will enter the cell it is designed to attack and transfer its code into that cell. Slide 19 Some current uses & trials of gene therapy Introduction of genes for new blood vessels for clogged arteries Introduction of genes into the blood stream that interferes with the replication of HIV Human growth hormone for Dwarfism Insulin production Glowing mice Replacing the cytoplasm in eggs to produce healthier eggs Slide 20 Injecting genes into the blood stream which are then carried to the target cells by viral carriers where they unload their genetic material. This is then used by the body to produce proteins that fight the disease. For the next decade gene therapy will only be used on somatic cells (only effect the patient and not their offspring) You and bioethicists will have to decide the future!