1. Cell potential, animal cloning Can any cell develop into a complete organism? How do you clone a sheep?

2. Regenerating plants from a single cell

4. Testing the developmental potential of nuclei from differentiated cells (Needle prick can activate some eggs) (or suck out meiotic spindle with pipette) Does tadpole develop?

6. Copyright ©2003 by the National Academy of Sciences Gurdon, J. B. and Byrne, J. A. (2003) Proc. Natl. Acad. Sci. USA 100, 8048-8052 Fig. 2. (A) In Amphibia, nuclear transfer success declines rapidly as cells differentiate

7. Copyright ©2003 by the National Academy of Sciences Gurdon, J. B. and Byrne, J. A. (2003) Proc. Natl. Acad. Sci. USA 100, 8048-8052 Fig. 2. (A) In Amphibia, nuclear transfer success declines rapidly as cells differentiate

8. Donor nucleusStages reached Zygoteblastula, tadpole, adult Blastulablastula, tadpole, adult Tadpole intestineblastula, tadpole, adult Cultured skin cellblastula (differentiated) Success of nuclear transplant experiments

9. Donor nucleusStages reached Zygoteblastula, tadpole, adult Blastulablastula, tadpole, adult Tadpole intestineblastula, tadpole, adult Cultured skin cellblastula Blastula from aboveblastula, tadpole Could some genes be missing from differentiated skin cells?

10. Serial nuclear transfer improves the efficiency of cloning

11. Conclusions of nuclear transplantation experiments in amphibians: 1) Nuclei of differentiated cells have the potential to program development of many or all cells. (Therefore, the nuclei must contain all the necessary genes.)

12. Conclusions of nuclear transplantation experiments in amphibians: 1) Nuclei of differentiated cells have the potential to program development of many or all cells. (Therefore, the nuclei must contain all the necessary genes.) 2) The capacity of nuclei to direct development is restricted as they differentiate. 3) The ooctye cytoplasm can reprogram differentiated nuclei to allow them to direct complete development.

13. 07_37_Protein.produc.jpg Nuclear effect – ability of genes to be expressed Cytoplasm effects – signals to nucleus (transcription factors, signaling pathways)

14. 08_14_chromatin.struc.jpg Genes need to be accessible to be expressed

15. 05_30_histone tails.jpg Histone modification state may limit gene expression potential

16. DNA methylation state may limit gene expression potential

17. Cleavage divisions in the frog Xenopus animal vegetal Blastula Why do frogs from nuclear transplant experiments often arrest at the blastula stage?

18. The mid-blastula transition (after about 12 rounds of cleavage) Growth phases added to Cell Cycle Embryo starts transcribing its own RNA

19. Cloning frogs from a single nuclear donor

20. 08_02_genetic.instruction.part2 Cloning mammals from adult cells (also activates egg)

21. (from Nature 405: 800-802)

23. NY Times, June 3, 2006 Clones' Debut Is a Test of Genetics, and Bettors' Wits By BILL FINLEY Anyone trying to select a winner at the mule races this weekend in Winnemucca, Nev., will no doubt have a hard time choosing between Idaho Gem and Idaho Star. They may have different names, but they are not necessarily different mules. Idaho Gem and Idaho Star are clones. They are two of three mules who were born in 2003 as the result of a cloning project at the University of Idaho and Utah State University. Leased from the University of Idaho for racing by Don Jacklin, an Idaho businessman, Gem and Star will make their first career starts today in separate 350-yard elimination races. The top eight finishers, based on time, will meet in tomorrow's final. There is parimutuel wagering on the races. Jacklin is confident the two mules will make the final, which could be part mule race, part science experiment. Who is faster, Idaho Gem or Idaho Star, or will they cross the finish line in unison? "Genetically, they should have equal ability," Jacklin said. "But you have to factor in the environmental effects. They can make a big difference." Jacklin counts his identical twin among the reasons that he is interested in the genetic sciences. He put up $400,000 to help create the clones. Dr. Gordon L. Woods, one of the veterinary scientists involved in the project, mated the parents of a champion mule named Taz and took cells from the resulting fetus. The cells were implanted in surrogates. Mules are a cross between female horses and male donkeys and are typically sterile.

24. http://brownfieldagnews.com/2010/08/25/iowa-grand-champion-steer- was-clone-of-2008-champ/ Iowa grand champion steer was clone of 2008 champ August 25, 2010 by Ken Anderson Ken Anderson 4-H livestock competitions have had their share of controversies over the years. Now 4-H officials have a potential new issue to deal with. DTN reports that the 2010 Iowa grand champion steer was a clone of the animal that won the same event in 2008. The crossbred steer was shown by Tyler Faber of Sioux Center, Iowa. According to the DTN report, Faber’s father, David Faber, is president of Trans Ova, a livestock production company in Sioux Center. The cloned steer was produced by Bovance, a joint venture between Trans Ova and the cloning firm ViaGen. Iowa 4-H livestock superintendent Mike Anderson says show officials found out the animal was cloned on Friday, after the steer had won the grand champion award two days earlier. Anderson says 4-H has no rule preventing clones from being shown—and he doesn’t think 4-H would create a rule on cloning because there’s no way for them to determine whether an animal is a clone. …

25. http://brownfieldagnews.com/2010/08/25/iowa-grand-champion-steer- was-clone-of-2008-champ/ Iowa grand champion steer was clone of 2008 champ August 25, 2010 by Ken Anderson Ken Anderson … According to the DTN report, livestock shows have not addressed issues with cloned cattle or hogs, but the status of clones is an issue in the horse industry. Thoroughbred and quarter horse racing associations have bans on cloned animals. By the way, the joint venture firm, Bovance, also paid the record 45- thousand dollar winning auction bid at the Iowa State Fair’s sale of champions to ensure the company would get to keep the cloned steer. A Bovance spokesman said the company bought the steer to keep in place the voluntary industry measure against meat from cloned animals going into the food supply for the general public.

26. Would it be ethical to clone humans? If so, for what purpose?