Advanced Protocols for Animal Transgenesis

This laboratory manual, published in cooperation with the International Society for Transgenic Technology (ISTT), provides almost all current methods that can be applied to the creation and analysis of genetically modified animals. The chapters have been contributed by leading scientists who are actively using the technology in their laboratories. Based on their first-hand experience the authors also provide helpful notes and troubleshooting sections.

Topics range from standard techniques, such as pronuclear microinjection of DNA, to more sophisticated and modern methods, such as the derivation and establishment of embryonic stem (ES) cell lines, with defined inhibitors in cell culture medium. In addition, related topics with relevance to the field are addressed, including global web-based resources, legal issues, colony management, shipment of mice and embryos, and the three R's: refinement, reduction and replacement.

Contenu

Karen S. Canady: Patent and licensing issues in transgenic technology.- Lluís Montoliu: Global Resources: Including Gene Trapped ES Cell Clones: Is Your Gene Already Knocked Out?.- Eduardo Moltó, Cristina Vicente-García and Lluís Montoliu: Designing Transgenes for Optimal Expression.- Thomas L. Saunders: Gene Targeting Vector Design for Embryonic Stem Cell Modifications.- Thomas J. Fielder and Lluis Montoliu: Transgenic Production Benchmarks.- Katja Becker and Boris Jerchow: Generation of Transgenic Mice by Pronuclear Microinjection.- Séverine Ménoret, Séverine Remy, et al : Generation of Transgenic Rats using Microinjection of Plasmid DNA or Lentiviral vectors.- Almudena Fernández, Diego Muñoz and Lluís Montoliu: Generation of Transgenic Animals by Use of YACs.- Michael G. Zeidler, Margaret L. Van Keuren, and Thomas L. Saunders: BAC Transgenes, DNA Purification, and Transgenic Mouse Production.- Carlos Lois: Generation of Transgenic Animals with Lentiviral Vectors.- Aron Geurts, Lajos Mates, Darius Balciunas: Vertebrate Transgenesis by Transposition.- Karen M. Chapman et al.: Rat Spermatogonial Stem Cell Mediated Gene Transfer.- Sayaka Wakayama, Nguyen Van Thuan and Teruhiko Wakayama: Mouse Cloning by Nuclear Transfer.- Elizabeth D. Hughes and Thomas L. Saunders: Gene Targeting in Embryonic Stem Cells.- Wojtek Auerbach and Anna B. Auerbach: The Importance of Mouse ES Cell Line Selection.- Marina Gertsenstein: Tetraploid Complementation Assay.- Elizabeth Williams et al.: Combining ES cells with Embryos.- Kristina Nagy and Jennifer Nichols: Derivation of Murine ES Cell Lines.- Ping Li et al.: Rat Embryonic Stem Cell Derivation and Propagation.- Han Li et al.: Induced pluripotency: generation of iPS cells from mouse embryonic fibroblasts.- Anna B. Auerbach, Peter J. Romanienko, and Willie H. Mark: The Preparation and Analysis of DNA for Use in Transgenic Technology.- Karen Brennan: Colony Management.- B. Pintado and J Hourcade: Cryopreservation.- Shirley Pease: Shipment of Mice and Embryos.- J.M. Sztein, R.J. Kastenmayer, K.A. Perdue: Pathogen Free Mouse Rederivation by IVF, Natural Mating and Hysterectomy.- Jan Parker-Thornburg: Refinement, Reduction and Replacement.