Florin Lore, Alter Heike, Gröne Hermann-Josef, Szabowski Axel, Schütz Günther, Angel Peter
Division of Signal Transduction and Growth Control, Deutsches Krebsforschungszentrum, Heidelberg, Germany.
Genesis. 2004 Mar;38(3):139-44. doi: 10.1002/gene.20004.
Loss-of-function approaches by the Cre/loxP technology have provided powerful tools for functional analyses of genes of interest expressed preferentially in a particular tissue. Here we describe the generation of transgenic mouse lines expressing Cre recombinase under the control of the promoter/enhancer unit of the gene for the alpha2 chain of collagen type I (Col1alpha2). As an expression vector, we used a P1-derived artificial chromosome (PAC), which harbors approximately 100 kb carrying the col1alpha2 gene. The improved coding sequence of the Cre recombinase was introduced to replace the first exon of col1alpha2. Cre expression was determined by immunohistochemistry and Cre-mediated onset of beta-galactosidase expression in ROSA26R-Cre reporter mice. In four analyzed transgenic lines, Cre recombinase was efficiently expressed during embryogenesis and in adult animals in cells of mesenchymal origin, such as dermal fibroblasts, mesenchymal cells of blood vessel walls, and cells in fibrous connective tissues surrounding internal organs.
通过Cre/loxP技术的功能缺失方法为优先在特定组织中表达的目标基因的功能分析提供了强大工具。在此，我们描述了在I型胶原α2链（Col1α2）基因的启动子/增强子单元控制下表达Cre重组酶的转基因小鼠品系的产生。作为表达载体，我们使用了一个源自P1的人工染色体（PAC），它携带约100 kb的包含col1α2基因的片段。引入改进的Cre重组酶编码序列以取代col1α2的第一个外显子。通过免疫组织化学以及ROSA26R-Cre报告基因小鼠中Cre介导的β-半乳糖苷酶表达的起始来确定Cre表达。在四个分析的转基因品系中，Cre重组酶在胚胎发育期间以及成年动物的间充质来源细胞中高效表达，如真皮成纤维细胞、血管壁间充质细胞以及围绕内脏器官的纤维结缔组织中的细胞。
