Principles of Cloning

Editorial Advisory Board

Foreword to the Second Edition

References

Foreword to the First Edition

Preface to the First Edition

In remembrance of Keith Henry Stockman Campbell (23 May 1954–5 October 2012)

List of Contributors

Part I: Basics

Chapter 1. Artificial Activation of Mammalian Oocytes for Cloning: Present Status and Future Perspectives

Introduction

Egg Activation Protocols

Common Activation Procedures for Mouse Eggs

Common Activation Procedures for Bovine and Porcine Eggs

Common Activation Procedures for Human Eggs

Conclusions

Acknowledgements

References

Chapter 2. Epigenetic Reprogramming of Somatic Nuclei via Cell Fusion

Introduction

Epigenetic Reprogramming of Somatic Cells by Hybridization with Embryonic Stem Cells

Epigenetic Reprogramming of Somatic Cells by Hybridization with Embryonic Germ Cells

Role of Cell Fusion In Vivo

Cloning by Somatic Cell Transplantation into Oocytes

Molecular Mechanisms of Epigenetic Reprogramming Mediated by Cell Fusion

Conclusion

References

Chapter 3. Nuclear Origins and Clone Phenotype: What Cloning Tells Us about Embryonic Development

Introduction

Aberrant Properties of Cloned Embryos

Effects of Developmental Stage of Donor Nucleus

Effects of Cell Type of Donor Nucleus

Effects of Donor Nucleus Cell Cycle Stage

Effects of Recipient Cell Type and Stage

Effects of Genotype and Sex of Donor Nucleus

Effects of Species Origin of Donor and Recipient – Challenges of Inter-Species Nuclear Transfer

Remediation of Aberrant Properties

Mysteries yet to be Solved

Acknowledgements

References

Part II: Methods

Chapter 4. Micromanipulation Techniques for Cloning

Introduction

Making Manipulation Tools

Microscopy and Equipment for Micromanipulation

Micromanipulation Procedures

Piezoelectric Assisted Nuclear Transfer

Technical Improvements

References

Chapter 5. Nuclear Transfer with Germ Cells: Germ Cell Cloning Contributes to Current Understanding of Genomic Imprinting in Mammals

Introduction

The Absence of Genomic Imprinting in Germ Cell (GC) Embryos

Detection of Erasure Process of Genomic Imprinting in Cloned Embryos Using Day 11.5 PGCs

Establishment of Genomic Imprinting Using Gonocyte Clone Embryos

Why is a Monoallelic Expression System of Imprinted Genes from Both Parental Alleles Essential for Mammalian Development?

Perspective

Acknowledgements

References

Chapter 6. Culture of Viable Mammalian Embryos In Vitro

Introduction

Dynamics of Embryo and Maternal Physiology

Metabolism of the Embryo

Impact of Intracellular Stress to the Embryo

Development of Sequential Embryo Culture Media

Inherent Problems of Co-Culture

Send in the Clones

Conclusions

References

Chapter 7. Genetic and Phenotypic Similarity Among Members of Mammalian Clonal Sets

Introduction

Definition of Cloning

Cytoplasmic Genetics

Epigenetic Effects

Uterine Effects

Neonatal Environment

Large Offspring Syndrome

Mutations

Cultural Inheritance

How Similar?

Summary and Perspective

References

Chapter 8. Advances in the Generation of Transgenic Domestic Species via Somatic Cell Nuclear Transfer

Introduction

Methods to Generate Transgenic Animals

Methods to Generate “Loss-Of-Function” Mutations Through SCNT

Additional Considerations

References

Chapter 9. Pregnancy and Neonatal Care of SCNT Animals

Overview of Cloning Efficiency

Health and Care of SCNT Cattle

Health and Care of SCNT Pigs

Health and Care of Other Species

Conclusion

Acknowledgements

References

Chapter 10. Donor Cell Type and Cloning Efficiency in Mammals

Developmental Potential of Germline Nuclei at Various Cell Cycle Stages

Developmental Potential of Somatic Cell Line Nuclei from Different Tissues

Acknowledgement

References

Chapter 11. Enhancing SCNT with Chromatin Remodeling Agents

Introduction

Establishment of a Concept for Direct Treatment of SCNT Embryos with Chemical Agents

Discovery of the Optimal Treatment of SCNT Embryos with Trichostatin A

SCNT using HDACi Treatment

The Possible Mechanism Underlying HDACi Treatment to Enhance Reprogramming

The Targets of HDACi to Enhance Nuclear Reprogramming

Why do Cloned Embryos Require HDACi Treatment for Better Genomic Reprogramming?

Further Studies of Agents that may Improve SCNT

Concluding Remarks

References

Chapter 12. Cell Cycle Regulation in Cloning

Introduction

Initial Events

Oocyte Development and the Cell Cycle

Cell Cycle Effects of Oocyte-Derived Cytoplast Recipients

Other Factors Related to the Recipient Cell Cycle Phase

Effects of Cell Cycle Combinations on Development

Effects of the Donor Cell Cycle Stage on Development

Summary

References

Chapter 13. Clone-Specific X-Linked Gene Repression Caused by Ectopic Xist Transcripts from the Active X Chromosome

Introduction

Aberrantly Expressed Genes In Pre-Implantation-Stage SCNT Embryos

Global Gene Expression Analysis and X-Chromosome Gene Repression in Cloned Embryos

Role of Xist in Pre-Implantation Embryos

Improvement of Cloning Efficiency Using Gene Knockout (KO) and Knockdown (KD) Animals

Why is Xist Expressed Ectopically in Scnt Pre-Implantation Embryos?

Xist-Independent Non-Random Errors in Cloned Embryos

Future Perspectives

References

Part III: Cloning by Species

Chapter 14. Cloning of Amphibia

Introduction and Background

Development of Nuclear Transfer Embryos Using Second Meiotic Metaphase Eggs

Immediate Changes in Transplanted Nuclei

Mechanisms of Nuclear Reprogramming by Meiotic Prophase Oocytes

Acknowledgements

References

Chapter 15. Cloning of Medaka Fish

Introduction

Study of Cloning in Medaka

Transfer of Adult Somatic Cell Nuclei into Diploidized Eggs

Successful Generation of Adult NTs

Mechanisms Requiring Investigation

Perspective

References

Chapter 16. Somatic Cell Nuclear Transfer in Zebrafish

Introduction

The Recipient Egg

The Donor Cells

Egg Activation Following Nuclear Transfer

Cloning Efficiency in Zebrafish

Conclusion and Closing Remarks

References

Chapter 17. Cloning of Mice

Introduction

Abnormalities Found in Cloned Mice

Where Are the Genomic Reprogramming Factors?

Improving Mouse Cloning by Technical Advances

Improvement of Mouse Cloning by Finding the Best Donor Cells

Combination of ntes Cell Approaches for Mouse Cloning

A New Tool for the Study of Basic Biology

Perspectives

References

Chapter 18. Cloning of Rabbits

Introduction

Background of Rabbit Embryo Embryology

State of the Art

Applications of Somatic Cell Nuclear Transfer in Rabbits

Appendix: Protocol for Rabbit Somatic Cell Nuclear Transfer

Acknowledgements

References

Chapter 19. Cloning Pigs by Somatic Cell Nuclear Transfer

Introduction

HIstory of SCNT in Swine

Applications of Genetically Engineered Pigs Produced Through SCNT

Culture of Pig Embryos

Uniqueness of SCNT Embryos

Reprogramming of Donor Cells

Methods to Improve the Efficiency of SCNT by Altering Epigenetic Marks

The Search for the Ideal Donor Cell Type

Future Direction

Acknowledgements

References

Chapter 20. A Historical Perspective of the Cloning of Cattle

Introduction

History

Cloning from Fetal Cells

Fetal Germ Line-Derived Cells

Fetal Somatic Cells

Cloning from Adult Somatic Cells

Aggregation of NT Embryos During Culture

Telomeres

Large-Offspring Syndrome

Food Safety of Milk and Meat Products Derived from the Use of Cloned Cattle for Milk or Meat Production

References

Chapter 21. Cloning of Canines

Introduction

History of Dog Cloning

Methodology of Canine Cloning

Postnatal Characteristics of cloneD Dogs

Applications of Nuclear Transfer

Acknowledgement

References

Chapter 22. Cloning of Equines

Procedures for Cloning Equines

Application of Cloning and Final Considerations

Acknowledgements

References

Chapter 23. Cloning Primates

Introduction

Overview of SCNT in Primates

Challenges in Primate SCNT

Note

Acknowledgements

References

Part IV: Applications

Chapter 24. Nuclear Transfer for Stem Cells

Introduction

Reprogramming Pluripotentiality by Nuclear Transfer

Reprogramming Pluripotentiality Using Transcription Factors

Human Somatic Cell Nuclear Transfer – What is the Difficulty?

The Future for SCNT Embryonic Stem Cells

References

Chapter 25. Current Research and Commercial Applications of Cloning Technology

Introduction

Cloning Research Animals

Previous and Present Obstacles to Commercial Cloning in Animal Agriculture

Summary

References

Chapter 26. Transgenic Cloned Goats and the Production of Recombinant Therapeutic Proteins

Introduction

Mammary Gland-Specific Transgenes for SCNT

Early Applications of SCNT for the Generation of Transgenic Goats

Oocyte Source, Cell Source, and SCNT Parameters

Generation of Transgenic Founders by Transfected SCNT

Production of Transgenic Goats by Adult SCNT

Detail Morbidity and Mortality

Transgenically Derived Recombinant Monoclonal Antibodies

Conclusion

Acknowledgements

References

Chapter 27. Commercializing Genetically Engineered Cloned Cattle

Research and Development

Cell Banking and Founder Animals

Animal Sourcing

Production Facilities

Quality and Risk

Animal Regulatory Oversight and Compliance in the United States

Conclusions

References

Chapter 28. Cloning Endangered Species

Introduction

Improving the Reproductive Potential of Endangered Animals

Pros and Cons of Using Somatic Cell Nuclear Transfer as a Bioremediation Tool

Zygotic Genome Activation Failure

Inbreeding

The Neglected Issue: Lack of Foster Mothers for Cloned Embryos

The State of the Art of Inter-Species Somatic Cell Nuclear Transfer for Multiplying Endangered Animals

Land Preservation

Establishing Biobanks for Endangered Animals

Improving Inter-Species Somatic Cell Nuclear Transfer

Removing Immunological Barriers to Embryo Transfer: Making the “Universal” Foster Mother

Conclusions

Acknowledgements

References

Part V: Complementary Technologies

Chapter 29. Genome Editing in Somatic Cells Using Zinc Finger Nucleases and Transcription Activator-Like Effector Nucleases

Setting the Stage for Genome Editing

The Power of the DSB

Making the Cut: ZFNs and TALENs

The Process of Creating a Modified Mammalian Somatic Cell

Acknowledgement

References

Chapter 30. Targeted Chromosome Elimination from ES-Somatic Hybrid Cells

Introduction

Homologous Recombination-Based Chromosome Engineering

Site-Specific Recombination

Systems to Introduce DNA Fragments

The Need for Techniques to Delete Large DNA Fragments

Chromosome Elimination Cassette

Conclusion

References

Part VI: SCNT as a Tool to Answer Biological Questions

Chapter 31. How Cell Division Facilitates Nuclear Reprogramming

Introduction

Nuclear Factors are Required for Reprogramming

Reprogramming Factors Localize to the Cytoplasm During Meiosis and Mitosis

Reprogramming Factors are Cell Type-Specific Regulators of Gene Expression

Transcriptional Reprogramming Depends on Mitotic Remodeling of the Nuclear Structure

Cell Division Correlates with Reprogramming to Induced Pluripotent Stem Cells

Nuclear Structure is Involved in the Regulation of Cell Type-Specific Gene Expression

Reprogramming of DNA Replication

Conclusion

Acknowledgements

References

Chapter 32. Proteomic Approach to the Reprogramming Machinery of the Mouse Oocyte

Introduction

Results

Discussion

Materials and Methods

Acknowledgements

References

Chapter 33. Biological Age of Cloned Animals

Introduction

Production of Embryos Cloned from Tissues Preserved for a Short Period

Cloning Regeneration of an Elite Bull from an Organ Frozen for more than a Decade Without Cryoprotectants

Concluding Remarks

References

Chapter 34. Mitochondrial DNA: Its Transmission from Gametes and Embryos

Introduction

The Mitochondrial Genome

Oxidative Phosphorylation

Mitochondrial DNA Replication

Segregation, Transmission, and Inheritance of Mitochondrial DNA

The Control of Mitochondrial DNA Replication During Development

How do Nuclear Transfer Embryos Control These Processes?

Mixing of Mitochondrial DNA Genotypes

Why Does the Donor Cell Mitochondrial DNA Persist?

Toxicity of Somatic Mitochondrial DNA

Is There a Relationship between Donor Cell and Recipient Oocyte Mitochondrial DNA that Affects Somatic Cell Nuclear Transfer Outcome?

Conclusion

Acknowledgement

References

Chapter 35. Interspecies Somatic Cell Nuclear Transfer

Introduction

Historical Perspective of Nuclear Transplantation (NT)

Interspecies Nuclear Transfer

Conclusions

References

Chapter 36. Epigenetics of Cloned Livestock Embryos and Offspring

Introduction

DNA Methylation Profiles in Cloned Embryos and Fetuses

Imprinted Gene Expression in Cloned Embryos and Fetuses

X-Chromosome Inactivation After Somatic Cloning

Telomere Length and Somatic Cloning

Conclusions and Perspectives

References

Chapter 37. Comparing SCNT-Derived ESCs and iPSCs

Introduction

Known Differences Between SCNT-ESCs and iPSCs

Conclusions

References

Chapter 38. Role of iPSC-Producing Factors in Pre-Implantation Embryos

Introduction

Expression Patterns and Functions of Reprogramming Factors in Pre-Implantation Embryos

Conclusions

References

Part VII: Ethical and Legal Affairs

Chapter 39. Genome Exchange in Human Oocytes

Introduction

Induced Pluripotent Stem Cells or Nuclear Transfer Embryonic Stem Cells?

A Source of Oocytes for Research

Human Oocytes Contain Reprogramming Activities

Genome Transfer to Prevent the Transmission of Mitochondrial Disease

Pronuclear Transfer in Zygotes

Genome Transfer in Unfertilized Human Oocytes

Acknowledgement

References

Chapter 40. Ethical Implications of Reproductive Cloning

Introduction

Reproductive Cloning

Nightmare Scenarios

Serious Ethical Concerns

Physiological Harms to Offspring

Psychological Harms

Social Harms

References

Chapter 41. An Overview of the Regulatory Considerations for Animal Cloning

Introduction

The US Process

Scientific Findings from other Competent Bodies

Statements Regarding Cloning from Scientific and Professional Organizations

Regulatory Status

What Does the Future Hold?

Future Developments and Expectations for Additional Research

Concluding Observations

References

Author Index

Subject Index