Transgenic mice

Transgenic mice constitute an entirely new dimension of mammalian molecular genetics. They allow us to observe genes at work in the intact organism. Virtually every sector of biomedical research is likely to be affected.     

Transgenic mice

Stable integration into the mouse genome of exogenous genetic information has become, over the past few years, a very potent approach for different aspects of biology. It is a common feature that the integrated exogenous gene (the transgene) is expressed properly both spatially and temporally. Constructing different lines of transgenic mice carrying various versions of a gene, therefore, permits cis acting DNA sequences involved in the specificity of expression to be defined, in the context of the developing animal. This in turn opens the way to a variety of experiments in which a given gene product is targeted to one or another cell type, thus offering some insight into the physiological role of this product. Such a strategy has been used, for example, to address the questions of the role of oncogenes in malignant transformation. The insertion of foreign DNA per se may disrupt the function of endogenous genes, thus creating an insertional mutation. The corresponding affected genes may subsequently be cloned, using the transgene as a tag. Finally, the ability to perform homologous recombination, recently demonstrated with embryonic stem cells that can colonize the germ line of a foreign embryo, should constitute in the near future a unique way to analyse in detail the functioning of the mammalian genome.     

Production of transgenic mice

A “transgenic” mouse is identified by the integration of a foreign DNA into its genome. Such animals serve as experimental systems for the study of gene expression and are often generated as models for human diseases. Direct microinjection of DNA into the male pronucleus of a mouse zygote has been the method most extensively used in the production of transgenic mice. Our method is subdivided into three sections: First, Preinjection, where the animals, the donor eggs, and the injection tools are presented. Second, Injection, where the egg handling and micromanipulation is described. Third, Postinjection, where the surgical transfer into pseudopregnant females completes the procedural sequence. Additional sections are provided to include the materials we use and to offer a series of technical tips that cover various aspects of the transgenic process.     

Cryopreservation of transgenic mice

Advances in cryopreservation enable one to freeze embryos without the use of a programmable freezing machine or complex protocols. These methods achieve high rates of survival when mouse embryos are frozen. Understanding the factors that influence the survival of cryopreserved embryos can aid troubleshooting and in adapting freezing strategies from mice to other species. Frozen stocks of transgenics can be maintained indefinitely in liquid nitrogen. Cryopreservation of these valuable animals not only protects them from environmental catastrophes, but also is an economical method of storing lines for future detailed analysis.     

The individuality of mice

Mutant mice simulating human CNS disorders are used as models for therapeutic drug development. Drug evaluation requires a coherent correlation between behavioral phenotype and drug status. Variations in behavioral responses could mask such correlations, a problem highlighted by the three-site studies of Crabbe et al. (1999) and Wahlsten et al. (2003a). Factors contributing to variation are considered, focusing on differences between individual animals. Genetic differences due to minisatellite variation suggest that each mouse is genetically distinct. Effects during gestation, including maternal stress, influence later life behavior; while endocrine exchanges between fetus and parent, and between male and female fetuses dependent on intrauterine position, also contribute. Pre and perinatal nutrition and maternal attention also play a role. In adults, endocrine cyclicity in females is a recognized source of behavioral diversity. Notably, there is increasing recognition that groups of wild and laboratory mice have complex social structures, illustrated through consideration of Crowcroft (1966). Dominance status can markedly modify behavior in test paradigms addressing anxiety, locomotion and aggressiveness, to an extent comparable to mutation or drug status. Understanding how such effects amplify the behavioral spectrum displayed by otherwise identical animals will improve testing.