Transgenesis Techniques

This fully updated and expanded edition details the transgenic techniques currently used to modify the genome. It explores classic procedures as well as cutting-edge practices involving microinjection, site-specific recombination systems, and cryopreservation.

One of the major challenges currently facing the scientific community is to understand the function of the multitude of protein-coding genes that were revealed when the human genome was fully sequenced. In Transgenesis Techniques: Principles and Protocols, Third Edition, experts in the field fully update and expand upon the previous edition in order to detail the transgenic techniques currently used to modify the genome. The volume explores classic procedures to genetically modify mice and other model organisms, as well as cutting-edge practices involving microinjection, site-specific recombination systems, cryopreservation, and many other topics. Written in the highly successful Methods in Molecular Biology™ series format, the chapters include brief introductions to their subjects, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and notes sections, highlighting tips on troubleshooting and avoiding known pitfalls.

Comprehensive and state-of-the-art, Transgenesis Techniques: Principles and Protocols, Third Edition is a comprehensive practical guide to the generation of transgenic animals and an invaluable source of information in any lab currently involved in transgenic techniques.

Provides state-of-the-art alongside the classics, i.e. classic techniques have been brought up to date, and are described along with modern state-of-the-art transgenesis technologies Covers a wide range of organisms, including Drosophila, C. elegans, Xenopus, zebrafish, rat and mouse Presents an accessible, user-friendly guide designed to inform and guide researchers currently involved in transgenic techniques and those new to the field Includes essential background information for scientists who work with genetically modified organisms but are not routinely involved in their generation Provides an essential blend of theory and practice Contributions by experienced authors from world renowned institutes Acts as an essential technical guide complete with lots of trouble shooting from researchers who have 'been there and done it'

Inhalt
Transgenesis in Various Model Systems.- Transgenesis in Drosophila melanogaster.- Transgenesis in Caenorhabditis elegans.- Transgenesis in Zebrafish with the Tol2 Transposon System.- Generation of Transgenic Frogs.- Pronuclear DNA Injection for the Production of Transgenic Rats.- Transgenesis in the Mouse.- Cell-Type-Specific Transgenesis in the Mouse.- Transgene Design and Delivery into the Mouse Genome: Keys to Success.- Overexpression Transgenesis in Mouse: Pronuclear Injection.- Gene-Targeting Vectors.- Gene Trap: Knockout on the Fast Lane.- Culture of Murine Embryonic Stem Cells.- Targeting Embryonic Stem Cells.- Generation of Chimeras by Microinjection.- Generation of Chimeras by Morula Aggregation.- Surgical Techniques for the Generation of Mutant Mice.- Site-Specific Recombinases for Manipulation of the Mouse Genome.- Cre Transgenic Mouse Lines.- Large-Scale Mouse Mutagenesis.- Dedicated Mouse Production and Husbandry.- Biological Methods for Archiving and Maintaining Mutant Laboratory Mice. Part I: Conserving Mutant Strains.- Biological Methods for Archiving and Maintaining Mutant Laboratory Mice. Part II: Recovery and Distribution of Conserved Mutant Strains.