The generation of genetically modified mice is absolutely crucial to gene function studies today, primarily because mice are genetically similar to man and because gene function studies in mice are in the context of a whole organism, making them particularly useful. In Transgenic Mouse Methods and Protocols, Second Edition, expert research explore current advances in the field through detailed laboratory protocols. Chapters provide a general introduction outlining how to deal with mice and how to generate transgenic mouse models, explore the generation of conditional and induced knockout and transgenic mice, and offer alternative routes to studying gene function in mice. Composed in the highly successful Methods in Molecular Biology™ series format, each chapter contains a brief introduction, step-by-step methods, a list of necessary materials, and a Notes section which shares tips on troubleshooting and avoiding known pitfalls. Comprehensive and state-of-the-art, Transgenic Mouse Methods and Protocols, Second Edition is the ideal guide for all researchers interested in the latest information about the production and analysis of transgenic and knockout mice
CONTENT
Introduction: Strategies for developing genetically modified mice -- Genetic Modification of the Mouse; General Technology: Pronuclear and Blastocyst Injection -- Generation of chimeras by aggregation of embryonic stem cells with diploid or tetraploid mouse embryos -- Cryopreservation of mouse spermatozoa and In vitro fertilization -- Autopsy and histological analysis of the transgenic mouse -- Transgene design -- Inducible Transgenic Mouse Models -- Lentiviral transgenesis -- Transgenesis in Mouse Embryonic Stem Cells -- Engineering BAC Reporter Gene Constructs for Mouse Transgenesis -- Targeting Vector Construction Through Recombineering -- Generating Conditional Knockout Mice -- Hypomorphic mice -- MICER Targeting Vectors for Manipulating the Mouse Genome -- Knockin approaches -- Generation of a series of knockin alleles using RMCE in ES cells -- Selection of targeted mutants from a library of randomly mutagenized ES cells -- Generation of genetically modified rodents using random ENU mutagenesis -- Bone marrow transplantations to study gene function in hematopoietic cells -- Adenovirus Mediated Gene Transfer
