In vivo Regulation of the Cell Fates Utilizing a Novel Inducible System of Protein Expression

利用新型蛋白质表达诱导系统体内调节细胞命运

• 批准号: 21591231
• 负责人: OTSU Makoto
• 金额: $ 2.91万
• 依托单位: The University of Tokyo
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2009
• 资助国家: 日本
• 起止时间: 2009 至 2011
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-21591231/
• 关键词: 造血幹細胞; レトロウイルスベクター; レンチウイルスベクター; 発現調節; 細胞老化

As planned, I constructed the inducible retroviral system that could allow rapid and dose-related regulation of expression of the target protein. It, however, turned out that the degradation-domain(DD)/Shield-1 system did have an intrinsic defect in itself, which was compatible with significant expression leak. I thus decided to change gears to the combined use of DD/Shield-1 and tetracycline-inducible system, the latter of which was originally developed by Dr. Bujard. I utilized so-called "All-in-One LV", the lentiviral system that was generated by my colleague to allow containing all the required elements for the system to work within a single cassette. Among the combination tested, I finally figured out that the system comprised of Tet-ON and DD/Shield-1 was the best performer so that it allowed the lowest leak and prompt expression upon dual stimulation. Although the overall progress remain behind the schedule due to the change of plans, I believe that the system is now ready to be tested in vivo, once the inducible target gene is introduced within this newly developed dual-regulation lentiviral vector.

按照计划，我构建了诱导型逆转录病毒系统，可以快速和剂量相关地调节靶蛋白的表达。然而，事实证明，降解结构域（DD）/Shield-1系统本身确实存在内在缺陷，这与显著的表达泄漏是相容的。因此，我决定改变齿轮DD/Shield-1和四环素诱导系统的联合使用，后者最初由Bujard博士开发。我使用了所谓的“All-in-One LV”，这是我的同事生成的慢病毒系统，允许包含系统在单个盒内工作所需的所有元素。在测试的组合中，我最终发现由Tet-ON和DD/Shield-1组成的系统性能最佳，因此它允许最低的泄漏和双刺激时的快速表达。尽管由于计划改变，总体进展仍落后于计划，但我相信，一旦将诱导型靶基因引入到这个新开发的双调节慢病毒载体中，该系统现在就可以进行体内测试。

项目成果

Deregulated intracellular signaling by mutated c-CBL in myeloid neonlasms.

髓系肿瘤中突变的 c-CBL 导致细胞内信号传导失调。

• 发表时间: 2010
• 期刊: Clin Cancer Res
• 影响因子: 11.5
• 作者: Ogawa S;et al.
• 通讯作者: et al.

先天性免疫不全症の遺伝子細胞治療

先天性免疫缺陷的基因和细胞疗法

• 发表时间: 2009
• 作者: 坂入和豊;田中由美子;権藤和美;畠中渚;國島伸治;安藤潔;松下弘道;宮地勇人;大津真
• 通讯作者: 大津真

抗CD40Lモノクローナル抗体投与を用いた骨髄混合キメリズム誘導による慢性肉芽腫症モデルマウスの治療

使用抗CD40L单克隆抗体诱导骨髓混合嵌合体治疗慢性肉芽肿病模型小鼠

• 发表时间: 2009
• 作者: 竹内康雄;他
• 通讯作者: 他

Transient activation of c-MYC expression is critical for efficient platelet generation from human induced nlurinotent stem cells.

c-MYC 表达的瞬时激活对于人类诱导性干细胞有效生成血小板至关重要。

• 发表时间: 2010
• 期刊: J Exp Med
• 影响因子: 15.3
• 作者: Takayama N;et al.
• 通讯作者: et al.

Towards the establishment of hematopoietic stem cell transplantation without the need for myeloablation

建立无需清髓的造血干细胞移植

• 发表时间: 2012
• 作者: Ishihara M;Heissig B;Nishida C;Rosenkvist J;Tashiro Y;Ohki M;Wanaka K;Tsuda Y;Okada Y;Nakauchi H;Hattori K;Makoto Otsu
• 通讯作者: Makoto Otsu