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Gene Clo Ning
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  • 2. Cloning
    • Different processes for duplicating biological material.
    • Diff types = technologies can be used for other purposes besides producing the genetic twin of another organism.
    • 1. DNA cloning,
    • 2. Reproductive cloning
    • 3. Therapeutic cloning .
  • 3. Recombinant DNA Technology or DNA Cloning
    • 1970s = common practice in MB labs today
    • “ Recombinant DNA technology," "DNA cloning," "molecular cloning“ or "gene cloning”
    • Transfer of a DNA fragment of interest from one organism to a self-replicating genetic element eg. bacterial plasmid.
    • The DNA of interest can then be propagated in a foreign host cell.
    • Scientists studying a particular gene often use bacterial plasmids to generate multiple copies of the same gene.
  • 4. Plasmids
    • Self-replicating circular DNA molecules
    • HGP = Plasmids and other types of cloning vectors are used by researchers to copy genes to generate sufficient identical material for further study.
    • Can carry up to 20,000 bp of foreign DNA
  • 5. How?
    • DNA fragment containing the gene of interest is cut from chromosomal DNA using RE.
    • United with a plasmid that has been cut with the same RE.
    • Fragment of chromosomal DNA + cloning vector = "recombinant DNA molecule."
    • The recombinant DNA can then be reproduced along with the host cell DNA.
  • 6. Other cloning vectors
    • Viruses
    • BACs = 100 – 300 kb
    • YACs. = 1 MB
    • Cosmids = up to 45 kb
  • 7. Reproductive cloning
    • Technology used to generate an animal that has the same nuclear DNA as another currently or previously existing animal.
    • Dolly. Roslin Institute.
    • Somatic cell nuclear transfer (SCNT), = transfer of GM from the nucleus of a donor adult cell to an egg whose nucleus, and GM has been removed.
    • The reconstructed egg containing the DNA from a donor cell must be treated with chemicals/ electric current to stimulate cell division.
    • Once the cloned embryo reaches a suitable stage, it is transferred to the uterus of a female host where it continues to develop until birth.
  • 8. Dolly the sheep
    • The first mammal to be cloned was put down by lethal injection Feb. 14, 2003.
    • Prior to her death, Dolly had been suffering from lung cancer and arthritis.
    • Studies showed that telomeres are shortened, a phenomenon that is associated with cellular aging
  • 9. Reproductive cloning
    • 1952 = tadpole.
    • The possibility of human cloning, raised when Scottish scientists at Roslin Institute created Dolly ( Nature 385, 810-13, 1997),
    • Since Dolly, sheep, goats, cows, mice, pigs, cats, and rabbits all using nuclear transfer technology.
    • Attempts at cloning certain species such as monkeys, chickens, horses, and dogs, have been unsuccessful as some species
    • ? some may be more resistant to somatic cell nuclear transfer than others.
    • Process can be traumatic
  • 10. Therapeutic cloning
    • Also called "embryo cloning," production of human embryos for use in research.
    • Not to create cloned human beings, but to harvest stem cells that can be used to study human development and to treat disease.
    • Stem cells are extracted from the egg after it has divided for 5 days.
    • Many researchers hope that one day stem cells can be used to serve as replacement cells to treat heart disease, Alzheimer's, CA and other diseases
  • 11. How to utilize recombinant tech?
    • Gene therapy = Tx genetic conds by introducing virus vectors that carry corrected copies of faulty genes into the cells of a host organism
    • GMO/GAO = Genes from different organisms that improve taste nutritional value or provide resistance
    • Sequencing genomes = Knowledge.
  • 12. How to utilize repro cloning
    • Can be used to develop efficient ways to reproduce animals with special qualities . Eg. Mass production of , drug-producing animals or animals that have been genetically altered to serve as models for studying human disease.
    • Could be used to repopulate endangered animals or animals that are difficult to breed. Eg. In 2001, the first clone of an endangered wild animal was born, a wild ox called a gaur. The young gaur died from an infection about 48 hours after its birth.
    • In 2001, scientists in Italy reported the successful cloning of a healthy baby endangered wild sheep, now living in Sardinia.
  • 13. How to utilize therapeu. cloning?
    • . May produce whole organs/ healthy cells that can replace damaged cells.
    • Eg transplants = stem cells would be used to generate an organ or tissue that is a genetic match to the recipient. In theory, the cloned organ could then be transplanted into the patient without the risk of tissue rejection.
    • If organs could be generated from cloned human embryos = the need for organ donation could be reduced.
  • 14. Problems of repro cloning
    • Reproductive cloning is expensive and highly inefficient.
    • > 90% of cloning attempts fail to produce viable offspring.
    • > 100 nuclear transfer procedures could be required to produce one viable clone.
    • Cloned animals tend to have compromised immune function and higher rates of infection, tumor growth, and other disorders.
    • Japanese studies have shown that cloned mice live in poor health and die early.
    • 1/3 calves born alive have died young, and many of them were abnormally large.
    • Don’t live long enough to generate good data.
    • In 2002, researchers at Cambridge reported that the genomes of cloned mice are compromised = certain of genes function abnormally.
  • 15.
    • Human cloning:
    • the most controversial debate of the decade. 
    • Aroused worldwide interest and concern
    • because of its scientific and ethical implications
    • Is it morally acceptable?
  • 16. Certain questions to be raised
    • Is self-engineering acceptable?
    • Will failures, eg. deformed offspring, be acceptable?
    • Will cloning lead to designer babies ?
    • Who is socially responsible for cloned humans?
    • Rights and legal protection?
  • 17.
    • 95% public say no to cloning,
    • but 95% of scientists say yes
  • 18. Reasons for cloning
    • Provide valuable research
    • Answer to infertility?
  • 19. Scenario 1
    • Scenario 1: A husband and wife who wish to have children but both are carriers of a lethal recessive gene.
    • “ Rather than risk the one in four chance of conceiving a child who will suffer a short and painful existence”
  • 20. Scenario 2
    • Scenario 2 : Parents of a terminally ill child are told that only a BMT can save the child's life:
    • “ With no donor available, parents attempt to clone a human being from cells of the dying child. If successful, the new child could be a match for BMT”
  • 21. Concerns of human cloning
    • Act of human arrogance = Playing God
    • Safety concerns = Due to the inefficiency of animal cloning (1-2 viable offspring for every 100 experiments) and the lack of understanding about reproductive cloning = unethical to attempt to clone humans.
    • Technology only in animals, in men = possibility of mutation/ bio damage?
  • 22.
    • Individuality and Uniqueness = Fear that a clone would not be an “individual” but merely a carbon copy.
    • Would it have a soul?
    • Clone would be constantly compared = burdened with oppressive expectations .
    • ????????????????????????????????
  • 23.
    • THE END OF ML429