Porret Andrée, Mérillat Anne-Marie, Guichard Sabrina, Beermann Friedrich, Hummler Edith
Transgenic Animal Facility, Faculty of Biology and Medicine, University of Lausanne, Switzerland.
Methods Mol Biol. 2006;337:185-205. doi: 10.1385/1-59745-095-2:185.
Analysis of genetically engineered mice is crucial for our understanding of the in vivo function of genes and proteins in the whole organism. This includes inactivation of a gene or the generation of specific mutations. The development of knockout and transgenic technologies in the mouse, therefore, represents a powerful tool for elucidating gene function, for modeling of human diseases, and potentially for the evaluation of drugs. In particular, conditional gene targeting applying the Cre/loxP-mediated recombination system is increasingly used to evaluate the role of the gene of interest in a cell-type-specific or even inducible manner. The experimental steps start with the characterization of the gene locus, followed by construction of a vector, gene targeting in ES cells, and establishment of mouse lines carrying the desired mutation. These are then bred to transgenic mice expressing Cre recombinase in a tissue-specific manner, thus allowing gene inactivation in a cell type of interest.
对基因工程小鼠的分析对于我们理解基因和蛋白质在整个生物体中的体内功能至关重要。这包括基因的失活或特定突变的产生。因此，小鼠基因敲除和转基因技术的发展是阐明基因功能、建立人类疾病模型以及潜在评估药物的有力工具。特别是，应用Cre/loxP介导的重组系统的条件性基因靶向越来越多地用于以细胞类型特异性甚至可诱导的方式评估感兴趣基因的作用。实验步骤首先是对基因座进行表征，然后构建载体、在胚胎干细胞中进行基因靶向以及建立携带所需突变的小鼠品系。然后将这些小鼠与以组织特异性方式表达Cre重组酶的转基因小鼠进行杂交，从而在感兴趣的细胞类型中实现基因失活。
