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

基因陷阱微阵列的开发及其在免疫病理学中的应用

• 批准号: 14570107
• 负责人: ISHIDA Yasumasa
• 金额: $ 2.18万
• 依托单位: NARA INSTITUTE OF SCIENCE AND TECHNOLOGY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2002
• 资助国家: 日本
• 起止时间: 2002 至 2003
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-14570107/
• 关键词: gene trap; DNA array; ES cells; mouse; knockout; regulatory T cells; autoimmunity; desease development mechanism; マイクロアレイ; データベース; NAISTrap

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.

DNA阵列能够在一次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基因靶向实验以进行基因功能的体内分析。随机基因片段进行分析。为了将上述两种实验系统的优点联合收割机结合起来，我们首先创建了约900个基因捕获的小鼠ES细胞done（http：//bsw3.aist-nara.ac.jp/kawaichi/naistrap.html），然后构建了来自破坏基因的cDNA阵列。使用这些阵列，我们鉴定了一个主要在小鼠脑中表达的新基因，并使用相应的ES细胞制备了该基因的破坏等位基因的纯合小鼠。对基因敲除小鼠的详细分析显示，基因捕获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阵列鉴定具有令人感兴趣的表达模式的基因或新基因候选物立即允许从具有 *rest序列的破坏的等位基因的ES细胞克隆产生敲除小鼠。

项目成果

E.Matsuda et al.: "Expression profiling with arrays of randomly disrupted genes in mouse embryonic stem cells leads to in vivo functional analysis"Proc.Nati.Acad.Sci.USA. 101・12. 4170-4174 (2004)

E.Matsuda 等：“小鼠胚胎干细胞中随机破坏基因的表达谱导致体内功能分析”Proc.Nati.Acad.Sci.USA 101・12 4170-4174 (2004)。

E.Matsuda et al.: "Expression profiling with arrays of randomly disrupted genes in mouse embryonic stem cells leads to in vivo functional analysis"Proc.Natl.Acad.Sci.USA. 101. 4170-4174 (2004)

E.Matsuda 等人：“用小鼠胚胎干细胞中随机破坏的基因阵列进行表达谱分析可进行体内功能分析”Proc.Natl.Acad.Sci.USA。

E.Matsuda, et al.: "Expression profiling with arrays of randomly disrupted genes in mouse embryonic stem cells leads to in vivo functional analysis"Proc.Natl.Acad.Sci.USA. 101. 4170-4174 (2004)

E.Matsuda 等人：“用小鼠胚胎干细胞中随机破坏的基因阵列进行表达谱分析可进行体内功能分析”Proc.Natl.Acad.Sci.USA。