1. Cloning…Cloning… Cloning

2. What do you think of cloning? www.mofirst.org/images/clone-tank.jpg www.guardian.co.uk/.../dinosaur-dna-recreation tdpri.com http://www.bamkapow.com/bk_images/mini_Me.jpg

3. What is a clone? Clone refers to genetically identical cells originating from one cell. Clone refers to genetically identical cells originating from one cell. Clones do NOT necessarily look alike Clones do NOT necessarily look alike Even identical twins are different due to the slight differences in environment they experience. Clones would be even more different as they would live in a very different environment and time

4. Brief history of cloning 1902: Determine chromosomes hold genetic info. 1902: Determine chromosomes hold genetic info. 1902: Embryo is divided in lab. 1902: Embryo is divided in lab. 1914: First successful nucleus transfer between 2 cells 1914: First successful nucleus transfer between 2 cells

5. Brief history of cloning 1952: Tadpoles cloned 1952: Tadpoles cloned 1953: Watson and Crick find the structure of DNA. 1953: Watson and Crick find the structure of DNA. 1962:Frogs cloned 1962:Frogs cloned

6. Brief history of cloning 1977: Mice created from single parent 1977: Mice created from single parent 1996: First animal cloned from adult cells is born, “Dolly” the sheep. 1996: First animal cloned from adult cells is born, “Dolly” the sheep.

7. Types of cloning DNA/gene cloning DNA/gene cloning Artificial twinning Artificial twinning Reproductive cloning Reproductive cloning Therapeutic cloning Therapeutic cloning

8. DNA/gene cloning A technique to reproduce DNA fragments or genes A technique to reproduce DNA fragments or genes Step 1: DNA fragment containing the gene Step 1: DNA fragment containing the gene Step 2: Fragment united with a “cloning vector”(Recombinant DNA molecule) Step 2: Fragment united with a “cloning vector”(Recombinant DNA molecule) Step 3: Recombinant DNA molecule introduced into a host cell Step 3: Recombinant DNA molecule introduced into a host cell Step 4: Recombinant DNA molecule can then be reproduced Step 4: Recombinant DNA molecule can then be reproduced Used by the HGP researchers to copy genes Used by the HGP researchers to copy genes www.biochem.arizona.edu/.../PLASMID.gif

9. Artificial Embryo Twinning Technology mimics the natural process of creating identical twins. Technology mimics the natural process of creating identical twins. In nature, twins occur just after fertilization of an egg cell by a sperm cell=> 1 egg, 2 genetically identical individuals In nature, twins occur just after fertilization of an egg cell by a sperm cell=> 1 egg, 2 genetically identical individuals Artificial embryo twinning occurs in a Petri dish instead of in the mother's body. Artificial embryo twinning occurs in a Petri dish instead of in the mother's body. Manually separating early embryo into individual cells, and then allowing each cell to divide and develop on its own. Manually separating early embryo into individual cells, and then allowing each cell to divide and develop on its own. Embryos are placed into a surrogate mother, where they are carried to term and delivered. Embryos are placed into a surrogate mother, where they are carried to term and delivered. Embryos are genetically identical. Embryos are genetically identical.

10. Reproductive cloning= SCNT Called "somatic cell nuclear transfer" (SCNT) Called "somatic cell nuclear transfer" (SCNT) Somatic cell (not sperm/egg cell): actually contains 2 complete sets of chromosome transfer Somatic cell (not sperm/egg cell): actually contains 2 complete sets of chromosome transfer Nuclear: The nucleus is like the cell's brain, contains DNA. Nuclear: The nucleus is like the cell's brain, contains DNA. Transfer: Moving an object from one place to another. Transfer: Moving an object from one place to another. So what does this really mean…… Transfer of genetic material from the nucleus of a donor adult cell to an egg whose nucleus, and thus its genetic material, has been removed. So what does this really mean…… Transfer of genetic material from the nucleus of a donor adult cell to an egg whose nucleus, and thus its genetic material, has been removed.

11. How does this work? Step 1: Isolate a somatic cell from an adult organism Step 1: Isolate a somatic cell from an adult organism Step 2: Transfer the nucleus from that cell to an egg cell from which the nucleus had been removed. Step 2: Transfer the nucleus from that cell to an egg cell from which the nucleus had been removed. Step 3: Treat the reconstructed egg containing the DNA from a donor cell with chemicals or electric current in order to stimulate cell division. Step 3: Treat the reconstructed egg containing the DNA from a donor cell with chemicals or electric current in order to stimulate cell division. Step 4: Implant the cloned embryo into a female host Step 4: Implant the cloned embryo into a female host A clone created through SCNT is not truly an identical clone of the donor animal. Only the clone's chromosomal or nuclear DNA is the same as the donor. A clone created through SCNT is not truly an identical clone of the donor animal. Only the clone's chromosomal or nuclear DNA is the same as the donor.

12. SCNT vs. Normal fertilization? Same result= a dividing ball of cells, called an embryo. Same result= a dividing ball of cells, called an embryo. Both contain two complete sets of chromosomes. Both contain two complete sets of chromosomes. BUT.. Where did those sets come from? Normal fertilization: sperm and egg both contain one set of chromosomes, combine, embryo gets 2 sets. Normal fertilization: sperm and egg both contain one set of chromosomes, combine, embryo gets 2 sets. SCNT: egg chromosomes are removed and replaced by chromosomes from a somatic cell, already has 2 sets of chromosomes SCNT: egg chromosomes are removed and replaced by chromosomes from a somatic cell, already has 2 sets of chromosomes

13. Therapeutic cloning Made in the same way as reproductive cloning except the embryo is not implanted Made in the same way as reproductive cloning except the embryo is not implanted Does not create a new genetically identical individual Does not create a new genetically identical individual Stem cells are harvested from the embryos produced Stem cells are harvested from the embryos produced The stem cells are used to grow different types of cells The stem cells are used to grow different types of cells VERY controversial process!!! VERY controversial process!!! Scientists feel that therapeutic cloning isn’t really cloning because the embryo is not allowed to live beyond 4-5 days Scientists feel that therapeutic cloning isn’t really cloning because the embryo is not allowed to live beyond 4-5 days But…if you believe that human life begins at conception, this young life is still a human being worthy of respect. But…if you believe that human life begins at conception, this young life is still a human being worthy of respect. Something to think about….. Does therapeutic cloning suggest that human life a just a commodity to be manipulated? Something to think about….. Does therapeutic cloning suggest that human life a just a commodity to be manipulated?

14. Reproductive vs. Therapeutic cloning

15. Other types of cloning Single cell organisms, like bacteria and fungi. (This includes fermentation processes for production of bread, beer, and wine.) Single cell organisms, like bacteria and fungi. (This includes fermentation processes for production of bread, beer, and wine.) Entire plant asexual replication Entire plant asexual replication Natural cloning occurs in sexual reproduction, when the embryo splits in two to produce twins. Natural cloning occurs in sexual reproduction, when the embryo splits in two to produce twins.

16. Meet Dolly…opening a can of worms In 1996, Dolly the sheep became the first vertebrate cloned from the cell of an adult animal. In 1996, Dolly the sheep became the first vertebrate cloned from the cell of an adult animal. Not only was this a remarkable scientific breakthrough but it immediately gained interest and concern from around the world on the future of cloning technology as it would effect humans. Not only was this a remarkable scientific breakthrough but it immediately gained interest and concern from around the world on the future of cloning technology as it would effect humans.

17. How did they do it…Step 1 Cell collected from a sheep’s udder (a somatic cell)

18. Step 2 Nucleus is removed from unfertilized egg of second sheep.

19. Step 3 Udder cell (somatic cell) is inserted (transferred) into egg with no nucleus.

20. Step 4 Insertion is successful.

21. Step 5   Electrical charge is supplied.   In order to start cell division an electrical charge or chemical reaction is needed

22. Step 6 Cells begin to divide.

23. Steps 7 and 8

24. How did they do it?

25. Summary of Dolly Experiment Dolly was cloned SCNT. Dolly was cloned SCNT. Somatic cells taken from a six-year-old white faced sheep and cultured in a lab. Somatic cells taken from a six-year-old white faced sheep and cultured in a lab. 277 cells then fused with 277 unfertilized eggs (each with the nucleus removed) 277 cells then fused with 277 unfertilized eggs (each with the nucleus removed) 29 viable reconstructed eggs survived and were implanted in surrogate Blackface ewes. 29 viable reconstructed eggs survived and were implanted in surrogate Blackface ewes. 1 gave birth to Dolly 1 gave birth to Dolly 0.361% survival 0.361% survival In nature between 33-50% of fertilized eggs develop. In nature between 33-50% of fertilized eggs develop.

26. Human Cloning and Ethics

28. Fertilization vs. cloning

29. Is it possible? YES!!! But… Just because we can do it, should we?

30. Let’s take a survey… Results of Time/CNN poll in 2001 of 1,015 adult Americans Results of Time/CNN poll in 2001 of 1,015 adult Americans P1-Question 1- clone animals: Good (29%), Bad (67%) P1-Question 1- clone animals: Good (29%), Bad (67%) P1-Question 2- clone humans: Good (7%), Bad (90%) P1-Question 2- clone humans: Good (7%), Bad (90%) P1-Question 3- reasons against cloning: Religion (34%), Distinctiveness (22%), Breeding a superior race (22%), Dangerous (14%) P1-Question 3- reasons against cloning: Religion (34%), Distinctiveness (22%), Breeding a superior race (22%), Dangerous (14%) P1-Question 4-personality: Yes (10%), No (74%) P1-Question 4-personality: Yes (10%), No (74%) P1-Question 5- clone yourself: Yes (5%), No (93%) P1-Question 5- clone yourself: Yes (5%), No (93%) P1-Question 6- timeline for a clone: 10 years (45%), 20 years (23%), 50+ years (10%), never (15%) P1-Question 6- timeline for a clone: 10 years (45%), 20 years (23%), 50+ years (10%), never (15%) P2- Question 1-cloning for organs: Yes (28%), No (68%) P2- Question 1-cloning for organs: Yes (28%), No (68%) P2- Question 2-cloning to save a life: Yes (21%), No (74%) P2- Question 2-cloning to save a life: Yes (21%), No (74%) P2- Question 3-cloning to help infertility: Yes (20%), No (76%) P2- Question 3-cloning to help infertility: Yes (20%), No (76%) P2- Question 4-delayed twins: Yes (10%), No(88%) P2- Question 4-delayed twins: Yes (10%), No(88%) P2- Question 5-clone on lost child: Yes (10%), No (88%) P2- Question 5-clone on lost child: Yes (10%), No (88%) P2- Question 6-same sex couple offspring: Yes( 10%), No (86%) P2- Question 6-same sex couple offspring: Yes( 10%), No (86%) P2- Question 7-superior race: Yes (6%), No (92%) P2- Question 7-superior race: Yes (6%), No (92%)

31. Pros/ Uses of cloning Cure human diseases- stem cells Cure human diseases- stem cells Create animals that are disease resistant Create animals that are disease resistant More consistent food products More consistent food products Save endangered species Save endangered species Replacing organs and other tissues Replacing organs and other tissues Infertility Infertility Replacing a lost child Replacing a lost child Creating donor people Creating donor people Harvest organs from animals Harvest organs from animals

32. Cons of/ problems with cloning Public perception- most of the population is against cloning Public perception- most of the population is against cloning Expensive Expensive Not efficient, low success rate Not efficient, low success rate Will divert resources and possibly delay cures Will divert resources and possibly delay cures Massive quantities of human eggs required =>Risking health and exploitation of women (coercion of egg donors) Massive quantities of human eggs required =>Risking health and exploitation of women (coercion of egg donors) Treats human life like a commodity Treats human life like a commodity Gateway to genetic manipulation and control of human beings Gateway to genetic manipulation and control of human beings Tumors, genetic defects in clones Tumors, genetic defects in clones Pre-mature ageing (genetic age) Pre-mature ageing (genetic age) Threat to a person’s uniqueness Threat to a person’s uniqueness Lack of knowledge Lack of knowledge

33. Deck stacked against us…. Sheep: 1 live birth out of 277 cloned embryos (0.4%) Mice: 5 live births out of 613 cloned embryos (0.8%) Pigs: 5 live births out of 72 cloned embryos implanted (7%) Goats: 3 live births out of 85 cloned embryos implanted (3.5%) Cattle: 30 live births out of 496 cloned embryos implanted (6%) (4.8%; 24 survived) Cat: 1 live birth out of 188 cloned embryos (0.5%) Rabbits: 6 live births out of 1852 cloned embryos (0.3%) (0.2%; 4 survived) This suggests that cloning is time consuming and expensive and does not yield a significant number of viable offspring

34. Is it legal? The United Nations, the Council of Europe, and the European Union all have laws on human cloning, but the laws are vague and do not differentiate between reproductive and therapeutic cloning Most countries do not have any specific legislation on cloning http://unesdoc.unesco.org/images/0013/001342/134277e.pdf

35. Cloning legality in the US Animal cloning controlled by FDA Animal cloning controlled by FDA 2001: Human Cloning Prohibition Act of 2001 (H.R. 2505) passed in the House July 31, 2001, not acted upon by the Senate 2001: Human Cloning Prohibition Act of 2001 (H.R. 2505) passed in the House July 31, 2001, not acted upon by the SenateHuman Cloning Prohibition Act of 2001 (H.R. 2505)Human Cloning Prohibition Act of 2001 (H.R. 2505) 2003: Human Cloning Prohibition Act of 2003 (H.R.234) passed the House February 27, 2003, not acted upon by the Senate 2003: Human Cloning Prohibition Act of 2003 (H.R.234) passed the House February 27, 2003, not acted upon by the SenateHuman Cloning Prohibition Act of 2003 (H.R.234)Human Cloning Prohibition Act of 2003 (H.R.234) 2007: The Human Cloning Prohibition Act of 2007 (H.R. 2560) introduced on June 5, 2007 and defeated in the House 2007: The Human Cloning Prohibition Act of 2007 (H.R. 2560) introduced on June 5, 2007 and defeated in the HouseThe Human Cloning Prohibition Act of 2007 (H.R. 2560)The Human Cloning Prohibition Act of 2007 (H.R. 2560) Prohibition on human cloning Prohibition on human cloning Criminal Penalty: Up to 10 years imprisonment Criminal Penalty: Up to 10 years imprisonment Civil penalty: Minimum 1 million dollar fine Civil penalty: Minimum 1 million dollar fine

36. Some ethical questions Are human clones individuals? Are human clones individuals? Do clones have parents? Do clones have parents? Who are they? Who are they? The cell donor? The cell donor? Nucleus donor? Nucleus donor? Surrogate? Surrogate? How old is a clone? Go by age of DNA or age of tissues? How old is a clone? Go by age of DNA or age of tissues? Do the benefits of cloning outweigh possible number of trials (possibly failed ones) that would have to occur with human embryos? Do the benefits of cloning outweigh possible number of trials (possibly failed ones) that would have to occur with human embryos? Is it ok to clone for outstanding individuals physically or mentally? For experiments? Is it ok to clone for outstanding individuals physically or mentally? For experiments? Who should have access to cloning technology – only people with good genes? Who should pay for the cloning? Should we be able to use this technology to design our children? Who does genetic information belong to? Should people with genetic diseases be cured? Should they have children