About Book

Transgenic animals ─ worms, flies, fishes, rodents and livestock ─ are the invaluable models that have allowed the scientific and medical community to study in depth the mechanisms of gene and genome function and complex disease phenotypes, to discover current and potential therapeutic approaches. Transgenic animals are routinely used in the laboratory as models in biomedical research. They are important tools for researching human disease, being used to understand gene function in the context of disease susceptibility, progression and to determine responses to a therapeutic intervention. The optimized techniques such as somatic cell nuclear transfer and cytoplasmic injection of exogenous DNA into zygotes are also the promising tools to generate transgenic livestock with special traits. This book elucidates new techniques and the applications in this field in a multidisciplinary approach. It elaborates on the different theories and concepts related to this area. It focuses on stem cells and animal transgenesis, senile systemic amyloidosis, transgenic animal technology, transboundary animal diseases, repetitive DNA and reproductive technologies. It also presents function measurements and recombineering of BAC DNA. The possible future directions and unresolved challenges are outlined. The topics included in this book are of utmost significance and are bound to provide incredible insights to the readers. It will serve as a valuable source of reference for those interested in this field and related branches.

Preface............................................................................................................v

Chapter 1 The Use of Reproductive Technologies to Produce Transgenic Goats..............................................................................................................1

Abstract...................................................................................................1

1. Introduction............................................................................................2

2. Pronuclear microinjection....................................................................4

3. Somatic cell nuclear transfer.............................................................11

4. Embryo transfer and pregnancy diagnosis....................................14

5. Conclusions..........................................................................................16

Acknowledgements.............................................................................16

Chapter 2 Repetitive DNA: A Tool to Explore Animal Genomes/Transcriptomes...........................................................................................21

1. Introduction..........................................................................................21

2. Organization of repetitive sequences..............................................23

3. Evolution and inter-species variation of repeat sequences........30

4. Functional significance of repetitive sequences............................31

5. Significance of repetitive sequences as marker.............................31

6. Bubalus bubalis genome....................................................................32

7. Repetitive sequences as molecular markers in bovid genome..33

8. Restriction Fragment Length Polymorphism (RFLP)...................34

9. Minisatellite Associated Sequence Amplification (MASA).........34

10. Technical approaches and methodologies......................................35

11. Applications in animal biotechnology............................................43

12. Concluding remarks...........................................................................44

Chapter 3 Transboundary Animal Diseases and International Trade............52

1. Introduction..........................................................................................52

2. Origins and catalysts of zoonoses and emerging diseases........53

3. Wildlife and their importance on international trade of

livestock´s products............................................................................55

4. Wildlife diseases monitoring systems and their importance for

international trade .............................................................................60

5. The one health concept: An holistic, integrative and

transdisciplinary paradigm for controlling, preventing and

monitoring TADs..................................................................................64

6. Conclusions..........................................................................................70

Chapter 4 Recent Advances and Applications of Transgenic Animal

Technology..................................................................................................81

1. Introduction..........................................................................................81

2. Microinjection .....................................................................................81

3. Sperm-mediated gene transfer.........................................................82

4. Embryonic stem cell-mediated gene transfer................................84

5. Somatic cell nuclear transfer............................................................87

6. Retrovirus-mediated gene transfer..................................................91

7. Germ line stem cell technique.........................................................93

8. Gene targeting.....................................................................................98

9. Gene silencing mediated by RNA interference..........................101

10. Zinc-finger nucleases – Gene targeting technique.....................103

11. Induced pluripotent stem cell technique.....................................104

12. Applications.......................................................................................108

13. Problems and prospects...................................................................110

14. Conclusion..........................................................................................111

Chapter 5 The Welfare of Transgenic Farm Animals.......................................119

1. Introduction........................................................................................119

2. Production-related disease related to extant breeding

technologies........................................................................................120

3. Ecological concerns raised by farm animal transgenesis..........127

4. Anthropocentric concerns raised by farm animal

transgenesis........................................................................................129

5. Using biotechnology to improve the welfare of farm

animals................................................................................................132

6. Meeting consumer expectations.....................................................136

7. Conclusion..........................................................................................138

Chapter 6 Spermatogonial Stem Cells and Animal Transgenesis.................146

1. Introduction........................................................................................146

2. Spermatogonial stem cells and spermatogenesis ......................147

3. Characterization of spermatogonial stem cells ........................149

4. SSC markers and differences among species .............................150

5. Isolation and in vitro culture of spermatogonial stem cells....152

6. SSCs transplantation and transgenesis ........................................154

7. Conclusions and perspectives .......................................................156

Chapter 7 Function Measurements of HLA-II Transgenic Pigs for Xenotransplantation................................................................................167

1. Introduction........................................................................................167

2. The expression of DP or DQ exogenes in HLA-DP or DQ

transgenic pigs reduced human-to-pig cellular responses.......168

3. The expression of DR exogenes in HLA-DR transgenic pigs

enhanced cyclosporine effect..........................................................171

4. Long-term survival of HLA-DR pig skin in SCID mice

after reconstitution with human PBMC and under short-term

immunosuppression..........................................................................174

5. Summary and conclusion................................................................176

6. Acknowledgements...........................................................................177

Chapter 8 Recombineering of BAC DNA for the Generation of Transgenic Mice............................................................................................................182

1. Introduction........................................................................................182

2. Overview of recombineering technology.....................................182

3. Recombineering materials for BAC modification.......................184

4. Recombineering methods and applications for BAC

modification........................................................................................185

5. Characterization of BAC transgenic mice....................................191

6. Utility of BAC transgenics..............................................................192

7. Conclusions........................................................................................194

Chapter 9 Transthyretin Amyloidosis in Aged Vervet Monkeys, as a

Candidate for the Spontaneous Animal Model of Senile Systemic

Amyloidosis..............................................................................................197

1. Introduction........................................................................................197

2. Animal models for TTR amyloidosis ..........................................198

3. Discussion and conclusion ............................................................204

4. Acknowledgment .............................................................................206

Citation......................................................................................................211

Index...........................................................................................................213

About Author

Drew Prentiss received his Ph.D. in Molecular Biology from Institute of Molecular Biology and Genetics, Leipzig, where he is currently working as a Research Fellow. His research interests are in genomic and phenotypic evolution of tumour cells under chemotherapeutic stresses and viro-immunological. Prentiss has authored a number of articles, journal papers, scientific papers and book chapters on transgenic animals. He has won several awards for his knowledge and contributions to science. He has also attended numerous conferences and symposiums.