Studies on vertebrate developmental genes by transposon-mediated gene trapping

转座子介导的基因捕获研究脊椎动物发育基因

• 批准号: 15201041
• 负责人: KAWAKAMI Koichi
• 金额: $ 22.8万
• 依托单位: National Institute of Genetics.
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (A)
• 财政年份: 2003
• 资助国家: 日本
• 起止时间: 2003 至 2005
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-15201041/
• 关键词: Molecular Biology; Developmental Genetics; Genome Biology; Zebrafish; Transposon; Transgenic fish; Gene expression; Gene trapping; エンハンサートラップ; インサーショナルミュータジェネシス; 器官形成; wntシグナル; 挿入変異; 転移酵素; トランスジェニック動物

(1)We injected a transposon-donor plasmid containing a To12 construct and mRNA encoding the transposase into zebrafish fertilized eggs and demonstrated that the transposon construct can be integrated in the genome of the germ cells very efficiently. This method is now widely used as a standard method to create transgenic fish.(2)We constructed the To12 construct containing a splice acceptor and the GFP reporter gene and created random integration of this construct in the zebrafish genome. When the splice acceptor trapped a transcript of an endogenous gene, the GFP gene is expressed in a pattern similar to that of the endogenous gene. This was the first successful demonstration of gene trapping in zebrafish.(3)We have been making efforts to distribute this useful methodology by publication of manuscripts, talks in international meetings, writing review articles, a training course and collaboration.(4)In the course of the transposon studies in zebrafish, we also found that the To12 transposon system is also active in mouse and Xenopus. To12 will be a useful gene transfer vector in these model vertebrates.

(1)我们将含有To12构建体和编码转座酶的mRNA的转座子供体质粒注射到斑马鱼受精卵中，并证明转座子构建体可以非常有效地整合到生殖细胞的基因组中。该方法现已广泛用作创建转基因鱼的标准方法。(2)我们构建了包含剪接受体和GFP报告基因的To12构建体，并在斑马鱼基因组中随机整合了该构建体。当剪接受体捕获内源基因的转录物时，GFP基因以与内源基因相似的模式表达。这是斑马鱼基因捕获的首次成功演示。(3)我们一直在努力通过发表手稿、在国际会议上演讲、撰写评论文章、培训课程和合作来传播这种有用的方法。(4)在斑马鱼转座子研究的过程中，我们还发现To12转座子系统在小鼠和爪蟾中也很活跃。 To12 将成为这些模型脊椎动物中有用的基因转移载体。

项目成果

Transposition of the To12 element, an Ac-like element from the Japanese medaka fish Oryzias latipes, in mouse embryonic stem cells.

To12 元件（一种来自日本青鳉鱼 Oryzias latipes 的 Ac 样元件）在小鼠胚胎干细胞中的转座。

• 发表时间: 2004
• 期刊: Genetics 166
• 作者: Kawakami;K.
• 通讯作者: K.

ゲノミクス・プロテオミクスの新展開(今中忠行監修)

基因组学和蛋白质组学的新进展（今中忠行指导）

• 发表时间: 2004
• 作者: Sunao Hara;Ayako Shirose;Chiyomi Miyajima;Katsunobu Ito;Kazuya Takeda;児玉晴男;川上浩一
• 通讯作者: 川上浩一

Clark, M.S., Edwards, Y.J.K., Peterson, D., Clifton, S.W., Thompson, A.J., Sasaki, M., Suzuki, Y., Kikuchi, K., Watabe, S., Kawakami, K., Sugano, S., Elgar, G., Johnson, S.L.: "Fugu ESTs : New resources for transcription analysis and genome annotation."Ge

克拉克，M.S.，爱德华兹，Y.J.K.，彼得森，D.，克利夫顿，S.W.，汤普森，A.J.，佐佐木，M.，铃木，Y.，菊池，K.，渡边，S.，川上，K.，菅野，S.

DNA transposons in vertebrate functional genomics

• DOI: 10.1007/s00018-004-4232-7
• 发表时间: 2005-03-01
• 期刊: CELLULAR AND MOLECULAR LIFE SCIENCES
• 影响因子: 8
• 作者: Miskey, C;Izsvák, Z;Ivics, Z
• 通讯作者: Ivics, Z

A transposon-mediated gene trap approach identifies developmentally regulated genes in zebrafish

• DOI: 10.1016/j.devcel.2004.06.005
• 发表时间: 2004-07-01
• 期刊: DEVELOPMENTAL CELL
• 影响因子: 11.8
• 作者: Kawakami, K;Takeda, H;Mishina, M
• 通讯作者: Mishina, M