Main description:

This volume provides readers with a historical foundation in standard techniques and a comprehensive update on the latest methods used in making gene-modified mice. The chapters in this book cover topics such as pronuclear microinjection in one-cell embryos; embryo transfer surgery; nuclear transfer and cloning; blastocyst microinjection; and cryobanking and recovery of genetically modified mice. Importantly, there are chapters devoted to the latest application of CRISPR technology, as well as the establishment of induced pluripotent stem cells. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls.

Comprehensive and authoritative, Transgenic Mouse: Methods and Protocols is a valuable resource for both novice and expert researchers who are interested in learning more about this evolving field.

Contents:

Preface...
Table of Contents...
Contributing Authors...

1. The History of Transgenesis
Thomas L. Saunders

2. Pronuclear Microinjection of One-Cell Embryos
Melissa A. Larson

3. Integrase-Mediated Targeted Transgenics through Pronuclear Microinjection
Ruby Yanru Chen-Tsai

4. In Vivo Validation of CRISPR Reagents in Pre-Implantation Mouse Embryos
Melissa A. Larson, Katelin A. Gibson, and Jay L. Vivian

5. Targeted Mutations in the Mouse via Embryonic Stem Cells
Marina Gertsenstein, Joffrey Mianne, Lydia Teboul, and Lauryl M.J. Nutter

6. Blastocyst Microinjection with Embryonic Stem Cells
Melissa A. Larson

7. Generation of Large Fragment Knock-In Mouse Models by Microinjecting into 2-Cell Stage Embryos
Bin Gu, Marina Gertsenstein, and Eszter Posfai

8. Embryo Transfer Surgery
Melissa A. Larson

9. Non-Surgical Embryo Transfer Protocol for use with the NSET (TM) Device
Barbara J. Stone

10. Nuclear Transfer and Cloning
Ling Liu

11. Sperm-Mediated Genetic Modifications
Marialuisa Lavitrano, Laura Farina, Maria Grazia Cerrito, and Roberto Giovannoni

12. Genotyping Genetically Modified (GM) Mice
Neeraj K. Aryal and Jan Parker-Thornburg

13. Nomenclature: Naming Your Gene-Modified Mouse
Alicia Valenzuela

14. Breeding Strategies for Genetically Modified Mice
Jan Parker-Thornburg

15. Strategies for Behaviorally Phenotyping the Transgenic Mouse
Kenneth E. McCarson

16. Cryobanking and Recovery of Genetically Modified Mice
Toru Takeo and Naomi Nakagata

17. Simple Transportation of Genetically Engineered Mice via Cold Storage Techniques
Hidetaka Yoshimoto, Toru Takeo, and Naomi Nakagata

18. Reprogramming of Primary Human Cells to Induced Pluripotent Stem Cells using Sendai Virus
Julia M. Draper and Jay L. Vivian