Full 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
The History of Transgenesis.- Pronuclear Microinjection of One-Cell Embryos.- Integrase-Mediated Targeted Transgenics through Pronuclear Microinjection.- In Vivo Validation of CRISPR Reagents in Pre-Implantation Mouse Embryos.- Targeted Mutations in the Mouse via Embryonic Stem Cells.- Blastocyst Microinjection with Embryonic Stem Cells.- Generation of Large Fragment Knock-In Mouse Models by Microinjecting into 2-Cell Stage Embryos.- Embryo Transfer Surgery.- Non-Surgical Embryo Transfer Protocol for use with the NSET™ Device.- Nuclear Transfer and Cloning.- Sperm-Mediated Genetic Modifications.- Genotyping Genetically Modified (GM) Mice.- Nomenclature: Naming Your Gene-Modified Mouse.- Breeding Strategies for Genetically Modified Mice.- Strategies for Behaviorally Phenotyping the Transgenic Mouse.- Cryobanking and Recovery of Genetically Modified Mice.- Simple Transportation of Genetically EngineeredMice via Cold Storage Techniques.- Reprogramming of Primary Human Cells to Induced Pluripotent Stem Cells using Sendai Virus.
