Generation of filggrin-deficient mice and analysis o differentiation and apoptosis of keratinocytes

有报道称，丝聚蛋白可能参与角质形成细胞的终末分化（凋亡）。为了研究角质形成细胞在体内的分化，我们计划产生丝状蛋白缺陷小鼠。聚丝蛋白由大约320个氨基酸和聚丝蛋白原的单位组成，聚丝蛋白原的单位由多个聚丝蛋白重复序列组成。为了产生聚丝蛋白缺陷小鼠，我们需要破坏包含聚丝蛋白原基因的起始密码子的区域，而不是聚丝蛋白的编码区域。我们成功地获得了一个BAC克隆含有小鼠微丝聚集蛋白原基因与人微丝聚集蛋白原cDNA作为探针。我们对该基因的重要区域进行了亚克隆，设计了靶向载体。目前，聚丝蛋白（filaggrin）^<+/-> ES细胞正在构建中。

It has been reported that filaggrin may be involved in the terminal differentiation (apoptosis) of keratinocytes. In order to study of keratinocyte differentiation in vivo, we are planning to generate filaggrin-deficient mice. Filaggrin is composed of approximately 320 amino acids and the unit of profilaggrin, which consists of multiple filaggrin-repeats. To generate filaggrin-deficient mice, we need to disrupt the region containing the initiation codon of the profilaggrin gene rather than the coding region of filaggrin. We succeeded to obtain a BAC clone containing the mouse profilaggrin gene with a human profilaggrin cDNA as a probe. We have subloned the important regions of this gene to design a targeting vector. At present, filaggrin^<+/-> ES cells are under construction.