DNA Technology and Genomics

1. DNA Technology and Genomics

2. Genetic Engineering/ DNA Technology • 3 types of Cloning Technologies: • Recombinant DNA Technology/ DNA Cloning • 2. Reproductive Cloning • 3. Therapeutic Cloning

3. Cloning the bovine growth hormone (BGH) gene

4. Recombinant DNA Technology/ DNA Cloning

5. Cloning a Gene • Cloning means making many copies of a gene

6. Recombinant DNA Technology/DNA Cloning 1. Remove the Gene of Interest 2. Cut the Plasmid DNA 3. Insert the Gene into the Bacterial Plasmid 4. Insert the Recombined Plasmid into the Bacterial Cell 5. Let the Bacteria Reproduce

7. 1. Remove the gene of interest (ex: BGH gene) using restriction enzymes Restriction enzymes cut DNA at specific sequences called palindromes . . .

8. Restriction Enzymes Leave “Sticky Ends”

9. Restriction Enzymes • The unpaired bases on the sticky ends form bonds with any complementary bases with which they come into contact

10. 2. Cut the Plasmid DNA with the Same Restriction Enzyme

11. 3. Insert the Gene of Interest into the Bacterial Plasmid (The bacterial plasmid is also cut with the restriction enzyme, leaving sticky ends)

12. 4. Insert the Recombined Plasmid into a Bacterial Cell

13. 5. Let the Bacteria Reproduce

14. Other Proteins Made by DNA Cloning: • Insulin for diabetics • Clotting factors for hemophiliacs

15. Basic Versus Applied Research Basic research • no profit motive or direct commercial application – generally government funded Applied research • immediate and profitable application – generally privately funded

16. Reproductive Cloning/Cloning Entire Organisms

17. Reproductive Cloning • Technology used to generate an animal that has the same nuclear DNA as another curretly or previously existing animal

18. Steps in Nuclear Transfer

19. Steps in Somatic Cell Nuclear Transfer

20. Results of Nuclear Transfer Experiments There were 277 failures before this nuclear transfer technique succeeded;

21. Reproductive Cloning Dolly was successfully born in 1997

22. Reproductive Cloning • Dolly was put to sleep at the age of 6 in 2003 • She was suffering from arthritis and a progressive lung disease • These are usually only seen in old sheep

23. Other Organisms Reproductively Cloned Mouse Cow Goat Mule Horse Rabbit Cat Pig Dog Rat Deer

24. Why Do Reproductive Cloning?

25. Have We Reproductively Cloned Humans?

26. Opposition and Support of Human Cloning

27. What do YOU think?

28. Therapeutic Cloning/ Embryo Cloning • Instead of cloning entire organisms, there is therapeutic cloning • Stem cells are induced to turn into specific tissue cells

29. Therapeutic Cloning

30. The Human Genome Project Sequenced the entire human genome

31. Goals of The Human Genome Project

32. Organisms With Genome Sequenced Fruit Fly Potato Zebra fish Rhesus Macaque

33. Many organisms have genome projects that have either been completed or will be completed shortly, including • : • Humans • Neanderthal • Haemophilus influenzae, a bacterium • Common House Mouse • Brown Rat • Common Chimpanzee • Rhesus Monkey • Domestic Chicken • Domestic Cat • Domestic Dog • Common fruit fly • Baker's yeast

34. Many organisms have genome projects that have either been completed or will be completed shortly, including • Red bread mold, • Thale Cress • Rice • Common Wheat • Corn • Poplar Tree • E. coli bacteria • SARS virus • Nematode worm • Zebra fish • Tomato • Potato • Western Honey Bee • Grapevine • Spanish flu virus

35. The Human Genome Project • These were used as model organisms in genetic studies • These model organisms contain genes that are the same as human genes

36. Gene Testing

37. DNA Chips

38. Pros and Cons of Gene Testing

39. Gene Therapy • Once the genetics are worked out, gene therapy can be researched • Replacing defective genes with functional ones • Germ linegene therapy in embryos • Somatic cell gene therapy in individual somatic cells in affected tissues

40. Gene Therapy • Germ Line Gene Therapy • Somatic Cell Gene Therapy

41. Gene Therapy • Non disease causing virus is genetically engineered with the functioning gene that is needed in SCID patients • The virus infects the immune cells and the immune cells get the functioning gene

42. Gene Therapy

43. Gene Therapy • Treatment for SCID (severe combined immunodeficiency) • Gene therapy in cells of the immune system

44. Somatic Gene Therapy • is not widely used • Is only for single gene disorders with cells that can be removed, engineered and then replaced in the body

45. Somatic Gene Therapy • The condition may still be passed to offspring, because somatic cell gene therapy does not treat all the cells in the body

46. Setbacks to Gene Therapy

47. Treat Cancer2. Gene Therapy to treat inherited form of blindness is successful! Recent Successes in Gene Therapy

48. Genetically Modified Crop Plants