1. Veterinary Medical Center SFT-ACT 2008 U NIVERSITY OF M INNESOTA CLONING DOGS AND CATS: WHERE DO WE STAND? Margaret V. Root Kustritz, DVM, PhD, DACT University of Minnesota

2. Veterinary Medical Center SFT-ACT 2008 U NIVERSITY OF M INNESOTA

3. Veterinary Medical Center SFT-ACT 2008 U NIVERSITY OF M INNESOTA CLONING - Nuclear transfer Nucleus from a single cell from the donor is transferred into an oocyte that has had its DNA removed, the new oocyte fertilized and matured in vitro, and the subsequent embryo transferred into a recipient dam at the morula or blastocyst stage.

4. Veterinary Medical Center SFT-ACT 2008 U NIVERSITY OF M INNESOTA CLONING - Nuclear transfer - Any donor cell can be used (skin biopsy from the ear)

5. Veterinary Medical Center SFT-ACT 2008 U NIVERSITY OF M INNESOTA After transfer of DNA, the single cell is stimulated to begin division IVM = in vitro maturation Immature oocyte released Low E2 / P4 environment Cumulus cells must be present

6. Veterinary Medical Center SFT-ACT 2008 U NIVERSITY OF M INNESOTA

7. Veterinary Medical Center SFT-ACT 2008 U NIVERSITY OF M INNESOTA SUCCESS RATES - Cats IVF and IVM - 50% development to the blastocyst stage Cats have been cloned (“Copy Cat”)

8. Veterinary Medical Center SFT-ACT 2008 U NIVERSITY OF M INNESOTA

9. Veterinary Medical Center SFT-ACT 2008 U NIVERSITY OF M INNESOTA SUCCESS RATES - Dogs IVF and IVM - 8-37% development to the blastocyst stage One lab has produced cloned dogs.

10. Veterinary Medical Center SFT-ACT 2008 U NIVERSITY OF M INNESOTA “Snuppy” = Seoul National University + puppy

11. Veterinary Medical Center SFT-ACT 2008 U NIVERSITY OF M INNESOTA CONCERNS - Disease of cloned animals - Early senescence? - Increased stillbirths - Birth defects - Embryo wastage - Cat return rate 1.0% - Dog return rate 0.1%

12. Veterinary Medical Center SFT-ACT 2008 U NIVERSITY OF M INNESOTA CONCERNS - Disease of cloned animals - Early senescence? - Increased stillbirths - Birth defects - Embryo wastage - Cat return rate 1.0% - Dog return rate 0.1%

13. Veterinary Medical Center SFT-ACT 2008 U NIVERSITY OF M INNESOTA CONCERNS - Embryo wastage* - Cat return rate 1.0% - Dog return rate 0.1% - Cattle return rate 11.0% - Mouse return rate 5.0% - Goat and sheep return rate 3.0% * Using adult cell for donor nucleus

14. Veterinary Medical Center SFT-ACT 2008 U NIVERSITY OF M INNESOTA CONCERNS - Expense - Public perception “Little Nicky”

15. Veterinary Medical Center SFT-ACT 2008 U NIVERSITY OF M INNESOTA CONCERNS - Expense - Public perception “Rainbow” and her clone, “Copy Cat”

16. Veterinary Medical Center SFT-ACT 2008 U NIVERSITY OF M INNESOTA COMMERCIAL AVAILABILITY - Projected cost = $50,000 / animal - Genetic Savings and Clone Website refers you to ViaGen – “Note that ViaGen has no plans to provide commercial dog or cat cloning services.”