Transgenesis techniques : principles and protocols (Methods in molecular biology, vol.180)

This revised and updated edition of a much-acclaimed classic presents a collection of readily reproducible techniques for the generation and analysis of transgenic animals, with emphasis on the manipulation of the mammalian genome. It concentrates on the creation and maintenance of genetically modified murine strains, providing access to the germline by both 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.

Topical reviews in transgenesis. Biomedical and agricultural applications of animal transgenesis. Transgenesis in invertebrate and lower vertebrate species. Gene transfer in drosophila. Transgenesis in the mouse : oocyte injection. Oocyte injection in the mouse. Alternative routes to the germline. Adenoviral infection. Retroviral infection. Transgenesis in the mouse : the ES cell route. In vitro isolation of murine embryonic stem cells. Production of chimeras derived from murine embryonic stem cells. Gene targeting strategies. Cre/loxP recombination system and gene targetingI. Cryopreservation of mouse lines. Cryopreservation of transgenic mouse lines. Ovarian tissue transplantation and cryopreservation : application to maintenance and recovery of transgenic and inbred mouse lines. Transgenesis in the rat. Transgenesis in the rat. Transgenesis in domestic species. Generation of transgenic livestock by pronuclear injection. Transgenic sheep from cultured cells. Characterization and analysis of transgenic strains. Analysis of transgenic mice. Index.