Development of highly effective gene knock-in procedure using neural crest-specific gene expression machinery

使用神经嵴特异性基因表达机制开发高效的基因敲入程序

• 批准号: 22700449
• 负责人: SATO Takahiro
• 金额: $ 2.58万
• 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: 2010
• 资助国家: 日本
• 起止时间: 2010 至 2011
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-22700449/
• 关键词: 発生工学

To identify the neural crest cell-specific enhancer region as well as transcription factors that bind to the enhancer region, we conducted luciferase assay using several upstream promoter regions on endothelin type-A receptor gene locus. Consequently, we found that about 30 bases as a critical enhancer sequence that consisted of presumable tandem binding sites of two transcription factors. In vivo experiments in transgenic mice exhibited an insufficiency of this enhancer sequence for neural crest cell-specific forced expression of LacZ gene. These results suggest that a possible coordination with other region may be required to proper gene expression in neural crest cells.Adenoviral transduction in fertilized eggs exhibited toxicity for their later normal development, whereas strong expression of LacZ was observed. Therefore, the important issue of viral toxicity remained to be overcome.

为了鉴定神经嵴细胞特异性增强子区域以及与增强子区域结合的转录因子，我们使用内皮素A型受体基因座上的几个上游启动子区域进行了荧光素酶测定。因此，我们发现大约 30 个碱基作为关键增强子序列，由两个转录因子的可能串联结合位点组成。转基因小鼠的体内实验表明，该增强子序列对于神经嵴细胞特异性强制表达 LacZ 基因来说是不足的。这些结果表明，神经嵴细胞中的正确基因表达可能需要与其他区域的协调。受精卵中的腺病毒转导对其后来的正常发育表现出毒性，而观察到LacZ的强表达。因此，病毒毒性的重要问题仍有待克服。

项目成果

Involvement of the endothelin-1/endothelin receptor type-A signaling in determination of the mandibular identity in mouse craniofacial development

内皮素-1/内皮素受体 A 型信号传导参与小鼠颅面发育中下颌特征的确定

• 发表时间: 2010
• 作者: Sato T;Kurihara Y;Asai R;Kawamura Y;Tonami K;Uchijima Y;Heude E;Ekker M;Levi G;Kurihara H
• 通讯作者: Kurihara H

Identification and developmental analysis of endothelin type-A expressing cells in the mouse kidney

小鼠肾脏A型内皮素表达细胞的鉴定及发育分析

• 发表时间: 2011
• 期刊: Gene Expr Patterns
• 影响因子: 1.2
• 作者: Kitazawa T;Sato T;Nishiyama K;Asai R;Arima Y;Uchijima Y;Kurihara Y;Kurihara H
• 通讯作者: Kurihara H

Endothelin type-A receptor expression defines a distinct subpoplation within the first heart field contributing to chamber myocardium

内皮素 A 型受体表达定义了第一心区内对心室肌有贡献的独特亚群

• 发表时间: 2010
• 作者: Asai R;Kurihara Y;Fujisawa K;Sato T;Kawamura Y;Kokubo H;Tonami K;Nishiyama K;Uchijima Y;Miyagawa-Tomita S;Kurihara H
• 通讯作者: Kurihara H

Identification and developmental analysis of endothelin receptor type-A expressing cells in the mouse kidney

• DOI: 10.1016/j.gep.2011.04.001
• 发表时间: 2011-10-01
• 期刊: GENE EXPRESSION PATTERNS
• 影响因子: 1.2
• 作者: Kitazawa, Taro;Sato, Takahiro;Kurihara, Hiroki
• 通讯作者: Kurihara, Hiroki

Transcription regulation of non-coding RNA Evf-2 in the Endothelin-1/Dlx5/Dlx6 pathway

Endothelin-1/Dlx5/Dlx6 通路中非编码 RNA Evf-2 的转录调控

• 发表时间: 2010
• 作者: Uchijima Y;Kurihara Y;Sato T;Fujisawa K;Kushiyama S;Kurihara H
• 通讯作者: Kurihara H