The Curious Case of the Carbon Copy Kitty

The Curious Case of the Carbon Copy Kitty by Nancy M. Boury Department of Animal Science Iowa State University, Ames, IA

The Star 2 Image used by permission from Macmillan Publishers Ltd: Nature (Shin, et

Part I: Kitty Troubles In each group - Person #1 Please tell your group the background on Patch and the problem faced by her TV show’s producers. 3

What does Clontronics propose? 4

So what is cloning? 5

Patch’s nucleus The Cloning Process 6

Video of Nuclear Transfer From: HHMI Biointeractive video site at http: //www. hhmi. org/biointeractive/video/index. html 7

CQ#1: Was the production of the clone a form of asexual or sexual reproduction? A. Asexual B. Sexual 8

CQ#2: What do you think? Can cloning produce a calico cat identical to Patch? A. Yes B. No 9

CQ#3: The cheek cells that are harvested are ___. A. Haploid, just like the rest of the other cells in Patch’s body. B. Triploid, because they are cheek cells. C. Diploid, just like the rest of the somatic cells in Patch’s body. D. Diploid, but half the chromosomes are in the cytoplasm, and half are in the nucleus. 10

CQ#4: Mr. Pseudofelis said if Patch was male, he could have cloned the cat using sperm without surgery. If he had used sperm, the cloned kitty would be ____. A. Identical to the original, they are clones after all. B. Diploid, but not identical. C. Haploid, but identical in appearance to the male sperm donor. D. Haploid, assuming it completed development. 11

Part II: Work In Progress Person #2: Please summarize the new developments in the cloning of Patch. 12

Patch’s nucleus The Cloning Process Fluffy 13

CQ#5: Assuming Clontronics used the same method of cloning used for Dolly the sheep, what contribution did Fluffy make to this process? A. B. C. D. E. She was the DNA donor. She was the egg donor. She was the sperm donor. She was the incubator. Fluffy was not needed to clone Patch. 14

CQ#6: If Clontronics is using the same kitty-cloning process described by Shin et al. , should the producers expect three viable kittens because there are three implanted embryos? A. Yes B. No 15

CQ#7: Do clones develop faster than normal offspring? A. Yes B. No 16

CQ#8: Is $2 million really the going rate for a cloned cat? A. Yes B. No 17

Part III: What Happened? ? This is the kitty that Clontronics delivered… 18 Image used by permission from Macmillan Publishers Ltd: Nature (Shin, et al, 2002, 415, 859) copyright 2002, http: //www. nature. com.

Person #3: Explain this newest development to your group. 19

Clontronics is sued for breach of contract. In their defense, they produce a DNA fingerprint of the clone, Patch and Fluffy. 20

DNA Fingerprinting http: //www. dnai. org 21

Exhibit A 22 Table used by permission from Macmillan Publishers Ltd: Nature (hin, et

CQ#9: There are two different numbers listed for each marker tested because___. A. Cats are haploid. B. The scientists wanted to check their work. C. Cats are diploid. D. Each marker is found in both the nuclear and extranuclear DNA. 23

CQ#10: If cats were tetraploid (4 n) species, each of these markers would have at most how many different lengths when testing a single cat? A. B. C. D. E. One Two Three Four Scientists can’t currently test anything that isn’t haploid 24

CQ#11: Clontronics promised a cat that was genetically identical to Patches. Given the data in Exhibit A, was the clone genetically identical to Patches? A. Yes B. No 25

CQ#12: What color is present in Patch, but absent in her clone? A. B. C. D. Orange Black White Patch is identical to her clone 26 Images used by permission from Macmillan Publishers Ltd: Nature (Shin, et al, 2002, 415, 859) copyright 2002, http: //www. nature. com.

Patches is a calico cat. Is this relevant? How are the orange and black patches formed in a calico cat? 27

Lyonization Black Orange Random X inactivation Black Orange Barr Body Orange B After several rounds of cleavage O=orange B= black O B B Black O Barr Body 28

CQ#13: The expert witness explained the process of lyonization to the jury. Based on this explanation, the cell that was taken from Patch had what difference that caused the difference in phenotype between clone and original cat? A. It had different mitochondria. B. It had a Barr body with an inactivated white producing X chromosome. C. It had a Barr body with an inactivated black pigment producing X chromosome. D. It had an active Y chromosome. E. It had a Barr body with an inactivated orange pigment producing X chromosome. 29

CQ#14: If you were on the jury, how would you vote? A. In favor of Clontronics, they did produce a genetically identical clone. B. In favor of the TV executives, they were misled, so give them their $20 million. C. In favor of the TV executives, but only award $2 million. 30

Exhibit B: Patch, Fluffy and the clone. 31 Image used by permission from Macmillan

For further reading What is warm and fuzzy forever? With cloning, kitty. New York Times Feb 15, 2002 32

Image Credits Slides 2, 18, 25, and 31 Description: Photo of cats. Source: From a two-paneled figure entitled “Figure 1: Nuclear-donor cat, and cloned kitten with its surrogate mother. ” Panel (a) portrays the adult female that supplied the cumulus cells for the nuclear transplantation; panel (b) shows the surrogate mother and the cloned kitten. From: Shin, T. , Kraemer, D. , Pryor, J. , Liu, L. , Rugila, J. , Howe, L. , Buck, S. Murphy, K. , Lyons, L. , Wethusin, M. (2002) A cat cloned by nuclear transplantation. Nature 415: 859. doi: 10. 1038/nature 723. Link: http: //www. nature. com/nature/journal/v 415/n 6874/fig_tab/nature 723_F 1. html. Clearance: Reproduced with permission from Macmillan Publishers Ltd: Nature, copyright 2002. License #2743810036853. Slides 6 & 13 Description: Diagram of reproductive and therapeutic cloning. Author: en: converted to SVG by Belkorin, modified and translated by Wikibob. Link: http: //en. wikipedia. org/wiki/File: Cloning_diagram_english. svg#file Clearance: Image licensed under the Creative Commons Attribution-Share Alike 3. 0 Unported license. Slide 7 Description: Somatic cell nuclear transfer. Source: Howard Hughes Medical Institute (HHMI). Link: http: //www. hhmi. org/biointeractive/media/scnt_video-lg. mov Clearance: Used in accordance with the terms of use of HHMI at http: //www. hhmi. org/popups/copyright. html. Slide 22 Description: Table 1. Analysis of feline genetic markers. Source: Shin, T. , Kraemer, D. , Pryor, J. , Liu, L. , Rugila, J. , Howe, L. , Buck, S. Murphy, K. , Lyons, L. , Wethusin, M. (2002) A cat cloned by nuclear transplantation. Nature 415: 859. doi: 10. 1038/nature 723. Link: http: //www. nature. com/nature/journal/v 415/n 6874/fig_tab/nature 723_F 1. html. Clearance: Reproduced with permission from Macmillan Publishers Ltd: Nature, copyright 2002. License #2743810036853. Slide 27 Description: Diagram of lyonization. Source: Nancy M. Boury, case author. Clearance: Used with permission. 33