Transgenic Animal Technology: A Laboratory Handbook

Transgenic animal technologies and the ability to introduce functional genes into animals have revolutionized our ability to address complex biomedical and biological questions. This well-illustrated handbook covers the technical aspects of gene transfer — from molecular methods to whole animal considerations — for important laboratory and domestic animal species. It describes methodologies as employed by leading laboratories and is a key resource for researchers, as well as a tool for training technicians and students. This second edition incorporates updates on a variety of genetic engineering technologies ranging from microinjection and ES cell transfer to nuclear transfer in a broad range of animal modeling systems.

Contains a comprehensive collection of transgenic animal and gene transfer methodsDiscusses background and introduction to techniques and animal systemsTeaches practical step-by-step protocolsFully revised with updates to reflect state-of-the-art technology and associated changes to date

Section One Overview. Introduction to transgenic animals. Section Two Transgenic animal production focusing on the mouse model. Transgenic animal production using DNA microinjection. Factors affecting transgenic animal production. Gene targeting in embryonic stem cells I: History and methodology. Gene targeting in embryonic stem cells II. Conditional technologies. Retrovirus-mediated gene transfer. Nuclear transfer technologies. Section Three Production of transgenic laboratory and domestic animal species. Production of transgenic rats. Production of transgenic rabbits. Production of transgenic fish. Production of transgenic poultry. Production of transgenic swine by DNA Microinjection. Production of transgenic ruminants by DNA Microinjection. Production of transgenic non-human primates. Retrovirus-mediated gene transfer into oocytes. Section Four: Molecular biology, analyses and enabling technologies Vector design for transgene expression. Analysis of transgene integration. PCR optimization for detection of transgenic integration. Evaluation of transgene expression. Control of gene expression (from standard gain/loss of function to anti-sense to PNAs). Enabling technologies: Cryopreservation of gametes. Enabling technologies: Embryo culture considerations. Databases, Internet Resources and Genetic Nomenclature.