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  1. 1. Cloning Dr. Abdelsalam Talafha, DVM Diplomate, American College of Theriogenologists
  2. 2. Cloning <ul><li>Production of genetically identical individuals that have identical nuclear DNA </li></ul>
  3. 3. Cloning Technologies <ul><li>Recombinant DNA technology </li></ul><ul><ul><li>DNA cloning </li></ul></ul><ul><ul><li>Molecular cloning </li></ul></ul><ul><ul><li>Gene cloning </li></ul></ul><ul><li>Reproductive cloning </li></ul><ul><li>Therapeutic cloning </li></ul><ul><ul><li>Embryo cloning </li></ul></ul>
  4. 4. DNA Cloning <ul><li>Transfer of a DNA fragment of interest from one organism to a self-replicating genetic element such as a bacterial plasmid </li></ul><ul><li>Plasmids </li></ul><ul><ul><li>Self-replicating extra-chromosomal circular DNA molecules, distinct from normal bacterial genome </li></ul></ul>
  5. 5. DNA Cloning - Uses <ul><li>Gene therapy </li></ul><ul><li>Genetic engineering of organisms </li></ul><ul><li>Genome sequencing </li></ul>
  6. 6. Reproductive Cloning <ul><li>A technology used to generate an animal that has same nuclear DNA as another currently or previously existing animal </li></ul><ul><li>Dolly </li></ul><ul><li>How Is Reproductive Cloning Done? </li></ul><ul><ul><li>Somatic cell nuclear transfer (SCNT) </li></ul></ul>
  7. 7. SCNT <ul><li>Starts with removal of polar body and chromosomes from an oocyte </li></ul><ul><ul><li>Enucleated oocyte </li></ul></ul><ul><li>Donor cell then inserted into perivitelline space of enucleated oocyte </li></ul>
  8. 8. SCNT <ul><li>Oocyte and donor cell are fused and activated by an electric pulse to begin cell division </li></ul><ul><li>Developed embryos transferred to surrogate females </li></ul><ul><li>Birth of an individual </li></ul>
  9. 17. SCNT <ul><li>Sources of somatic cells </li></ul><ul><ul><li>Cell from individual </li></ul></ul><ul><ul><li>Cells grown in culture </li></ul></ul><ul><ul><li>Frozen tissue </li></ul></ul>
  10. 18. Therapeutic Cloning <ul><li>Production of human embryos for use in research </li></ul><ul><li>Goal </li></ul><ul><ul><li>To harvest stem cells that can be used to study human development and to treat disease </li></ul></ul>
  11. 19. Therapeutic Cloning <ul><li>Stem cells </li></ul><ul><ul><li>Cells have ability to divide and give rise to both specialized cells and more stem cells </li></ul></ul><ul><li>Derived from </li></ul><ul><ul><li>Adults </li></ul></ul><ul><ul><li>Preimplantation embryos (Embryonic stem cells) </li></ul></ul>
  12. 20. Stem Cells <ul><li>Replacement cells to treat </li></ul><ul><ul><li>Heart disease </li></ul></ul><ul><ul><li>Alzheimer's </li></ul></ul><ul><ul><li>Cancer </li></ul></ul><ul><ul><li>Diabetes </li></ul></ul><ul><ul><li>Parkinson's disease </li></ul></ul><ul><ul><li>Spinal cord injury </li></ul></ul>
  13. 21. SCNT- Potential Biotechnological Applications <ul><li>Harvesting donor cells from transgenic animal </li></ul><ul><li>Genetic modification of cultured donor cells prior to nuclear transfer </li></ul><ul><ul><li>Producing genetically modified cloned offspring </li></ul></ul>
  14. 22. Genetically Modified Cloned Offspring <ul><li>Biotechnological applications </li></ul><ul><ul><li>Production of pharmaceuticals </li></ul></ul><ul><ul><li>Xenotransplantation </li></ul></ul><ul><ul><li>Study and eradication of human disease </li></ul></ul><ul><ul><li>Improvement of livestock </li></ul></ul>
  15. 23. Production Of Pharmaceuticals <ul><li>Gene expressing human coagulation factor IX introduced into ovine fetal fibroblasts </li></ul><ul><li>linked to another gene with a high level of expression in mammary gland </li></ul><ul><li>Protein expressed in milk </li></ul>
  16. 24. Production Of Pharmaceuticals <ul><li>Insulin for diabetes </li></ul><ul><li>Interferon for viral infections </li></ul><ul><li>Tissue plasminogen activator (which dissolves blood clots) </li></ul>
  17. 25. Xenotransplantation <ul><li>Aim: </li></ul><ul><ul><li>To develop animals whose organs will not cause an immunological response and destroy transplanted tissue when transferred to humans </li></ul></ul>
  18. 26. Xenotransplantation <ul><li>Pig organs </li></ul><ul><ul><li>Hearts, lungs, kidneys, liver </li></ul></ul><ul><ul><li>Neural tissue for Parkinson's </li></ul></ul><ul><ul><li>Islets cells for diabetes patients </li></ul></ul>
  19. 27. Study Human Disease <ul><li>Sheep model to investigate human cystic fibrosis </li></ul><ul><li>Cloned sheep used for drug testing and to evaluate new therapies </li></ul>
  20. 28. Improvement Of Livestock <ul><li>Cloning animal with excellent traits </li></ul><ul><li>Production of a large number of clones from high quality animals </li></ul><ul><ul><li>Allow overall genetic improvement of herd </li></ul></ul><ul><li>Repopulate endangered animals </li></ul>
  21. 29. Animal Cloning <ul><li>Sheep, cattle, goats, pigs, and mice </li></ul><ul><li>Cloning efforts in rabbits, rats, cats, dogs, and horses are ongoing </li></ul>
  22. 30. Risks Of Cloning <ul><li>Reproductive cloning expensive and highly inefficient </li></ul><ul><li>> 90% of cloning attempts fail to produce viable offspring </li></ul>
  23. 31. Risks Of Cloning <ul><li>High rate of fetal loss during pregnancy </li></ul><ul><li>Compromised immune function </li></ul><ul><li>Higher rates of infection </li></ul><ul><li>Tumor growth </li></ul><ul><li>Early neonatal death </li></ul><ul><li>Abnormally large at birth </li></ul><ul><li>Die mysteriously </li></ul>
  24. 32. Risks Of Cloning <ul><li>Cloned fetuses have abnormalities </li></ul><ul><ul><li>Abnormal placentation </li></ul></ul><ul><ul><li>Pregnancy toxemia </li></ul></ul><ul><ul><li>Hydroallantois </li></ul></ul>
  25. 33. Should Humans Be Cloned? <ul><li>Due to </li></ul><ul><ul><li>Inefficiency of animal cloning </li></ul></ul><ul><ul><li>Lack of understanding about reproductive cloning </li></ul></ul><ul><ul><li>Risks of cloning </li></ul></ul>
  26. 34. Should Humans Be Cloned? <ul><li>Unethical to attempt to clone humans </li></ul><ul><ul><li>Same problems would be expected in human cloning </li></ul></ul><ul><ul><li>We do not know how cloning could impact mental development </li></ul></ul>
  27. 35. Should Humans Be Cloned? <ul><li>High risk to health of fetus or infant and mother </li></ul><ul><li>Psychological risks for mother as a result of </li></ul><ul><ul><li>Late spontaneous abortions </li></ul></ul><ul><ul><li>Birth of a stillborn child </li></ul></ul><ul><ul><li>Birth of a child with severe health problems </li></ul></ul>