The past decade has witnessed a spectacular explosion in both the devel- ment and use of transgenic technologies. Not only have these been used to aid our fundamental understanding of biologic mechanisms, but they have also faci- tated the development of a range of disease models that are now truly beginning to impact upon our approach to human disease. Some of the most exciting model systems relate to neurodegenerative disease and cancer, where the availability of appropriate models is at last allowing radically new therapies to be developed and tested. This latter point is of particular significance given the current concerns of the wider public over both the use of animal models and the merits of using genetically modified organisms. Arguably, advances of the greatest significance have been made using mammalian systems―driven by the advent of embryonic stem-cell–based strategies and, more recently, by cloning through nuclear transfer. For this reason, this new edition of Transgenesis Techniques focuses much more heavily on manipulation of the mammalian genome, both in the general discussions and in the provision of specific protocols.
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The past decade has witnessed a spectacular explosion in both the development and the use of transgenic technologies. In Transgenesis Techniques: Principles and Protocols, Second Edition, authoritative researchers-including members of the Dolly team-take stock of these exciting developments to present a collection of readily reproducible techniques for the generation and analysis of transgenic animals, focusing on the manipulation of the mammalian genome. This second edition devotes much space to the creation of genetically modified murine strains, providing access to the germline both by conventional pronuclear injection and by retroviral and adenoviral infection. Extensive coverage is also given to the generation, maintenance, and manipulation of embryonic stem cell lineages, with protocols for both constitutive and conditional (Cre-lox or the Flp-frt systems) gene targeting. Additional chapters provide cutting-edge techniques for the cryopreservation of both male and female germlines, for the generation of transgenic sheep by nuclear transfer, and for the basic analysis of transgenic organisms.
Comprehensive and highly practical, this second edition of Transgenesis Techniques: Principles and Protocols updates and expands the acclaimed first edition with a wealth of easy-to-follow cutting-edge protocols and practical tips for producing transgenic animals and analyzing their integrated transgenes.
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