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Cloning

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