DEVELOPMENT OF THE GENE-TRAP MICROARRAY AND ITS APPLICATION TO IMMUNOPATHOLOGY

DNA阵列能够在单个实验中分析许多基因的表达模式。然而，在DNA阵列中找到感兴趣的基因后，有时必须进行劳动密集型的基因靶向实验，以进行基因功能的体内分析。用随机基因片段进行分析。为了结合上述两种实验系统的优点，我们首先创建了大约900个基因捕获的小鼠胚胎干细胞（http://bsw3.aist-nara.ac.jp/kawaichi/naistrap.html）and)，然后构建了来自中断基因的cdna阵列。利用这些阵列，我们确定了一个在小鼠大脑中主要表达的新基因，并使用相应的胚胎干细胞来产生小鼠纯合子，用于该基因的中断等位基因。对敲除小鼠的详细分析表明，基因诱捕载体完全消除了其整合位点下游的基因表达。因此，使用这种类型的DNA阵列识别具有有趣表达模式的基因或新候选基因，可以立即从具有*rest序列等位基因中断的胚胎干细胞克隆中产生敲除小鼠。

DNA arrays are capable of profiling the expression patterns of many genes in a single experiment. After finding a gene of interest in a DNA array, however, labor-intensive gene targeting experiments sometimes must be performed for the in vivo analysis of the gene function. With random gene segments are analyzed. In order to combine the benefits of the above two experimental systems, we first created about nine hundred gene-trapped mouse ES cell dones(http://bsw3.aist-nara.ac.jp/kawaichi/naistrap.html)and then constructed arrays of cDNAs derived from the disrupted genes Using these arrays, we identified a novel gene predominantly expressed in the mouse brain, and the corresponding ES cell done was used to produced mice homozygous for the disrupted allele of the gene. Detailed analysis of the knockout mice revealed that the gene trap vector completely abolished gene expression downstream of its integration site. Therefore, identification of a gene or a novel-gene candidate with an interesting expression pattern using this type of DNA arrays immediately allows the production of knockout mice from an ES cell clone with a disrupted allele of the sequence of *rest.