Production of gene Knock-down animal by introducing RNAi into spermatogonial stem cells.

通过将 RNAi 引入精原干细胞来生产基因敲除动物。

• 批准号: 20790302
• 负责人: TAKASHIMA Seiji
• 金额: $ 2.75万
• 依托单位: Kyoto University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Young Scientists (B)
• 财政年份: 2008
• 资助国家: 日本
• 起止时间: 2008 至 2009
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-20790302/
• 关键词: 疾患モデル動物; 遺伝子改変動物; 精子幹細胞; RNA interference

To assess whether RNAi mediated gene knock-down could be applied to germline stem cells, RNAi construct for EGFP knock-down was transduced to germline stem cells derived from EGFP transgenic mouse. Although lentivirus mediated RNAi transduction suppressed the EGFP expression to some extent in germline stem cells, the efficiency was not enough to produce the knock-down mouse. Meanwhile, by using RNAi technology developed in this project, it was shown that DNA methyl transferase I is indispensable for the survival of germline stem cells in vitro, suggesting that RNAi mediated knockdown is available for the endogenous genes.

为探讨RNAi介导的基因敲除在生殖系干细胞中的应用，将构建的绿色荧光蛋白敲除RNAi载体导入绿色荧光蛋白转基因小鼠的生殖系干细胞中。尽管慢病毒介导的RNAi转导在一定程度上抑制了生殖系干细胞中EGFP的表达，但其效率不足以产生击倒小鼠。同时，利用本项目开发的RNAi技术，表明DNA甲基转移酶I对生殖系干细胞的体外存活是必不可少的，表明RNAi介导的内源性基因敲除是可行的。

项目成果

Transmission distortion by loss of p21 or p27 cyclin-dependent kinase inhibitors following competitive spermatogonial transplantation

• DOI: 10.1073/pnas.0914448107
• 发表时间: 2010-04-06
• 期刊: PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA
• 影响因子: 11.1
• 作者: Kanatsu-Shinohara, Mito;Takashima, Seiji;Shinohara, Takashi
• 通讯作者: Shinohara, Takashi

Abnormal DNA Methyltransferase Expression in Mouse Germline Stem Cells Results in Spermatogenic Defects

• DOI: 10.1095/biolreprod.108.074708
• 发表时间: 2009-07-01
• 期刊: BIOLOGY OF REPRODUCTION
• 影响因子: 3.6
• 作者: Takashima, Seiji;Takehashi, Masanori;Shinohara, Takashi
• 通讯作者: Shinohara, Takashi

Characterization of laminin isoforms in human amnion

• DOI: 10.1016/j.tice.2007.09.001
• 发表时间: 2008-04-01
• 期刊: TISSUE & CELL
• 影响因子: 2.6
• 作者: Takashima, Seiji;Yasuo, Masanori;Nikaido, Toshio
• 通讯作者: Nikaido, Toshio

Phenotypic plasticity of mouse spermatogonial stem cells.

• DOI: 10.1371/journal.pone.0007909
• 发表时间: 2009-11-19
• 期刊: PloS one
• 影响因子: 3.7
• 作者: Morimoto H;Kanatsu-Shinohara M;Takashima S;Chuma S;Nakatsuji N;Takehashi M;Shinohara T
• 通讯作者: Shinohara T

Homing of mouse spermatogonial stem cells to germline niche depends on betal-integrin

小鼠精原干细胞归巢至种系生态位取决于β-整合素

• 发表时间: 2008
• 期刊: Cell Stem Cell 3
• 作者: Kanatsu-Shinohara;M et.al.
• 通讯作者: M et.al.