Development of the method for measurement of cellular dynamics by using single cell time course analysis.

开发使用单细胞时间过程分析测量细胞动力学的方法。

• 批准号: 20760534
• 负责人: HAKAMADA Kazumi
• 金额: $ 2.75万
• 依托单位: The University of Tokyo
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Young Scientists (B)
• 财政年份: 2008
• 资助国家: 日本
• 起止时间: 2008 至 2010
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-20760534/
• 关键词: 一細胞計測; 時系列解析; 細胞状態; 一細胞; 時系列; 遺伝子発現メカニズム; プロモータ活性

In this study, we elucidate the strong correlation between cell division and onset of gene expression by measuring their time course of HeLa cells, which were taransfected with EGFP (or Venus). In the next, we investigate the generality of this phenomenon by 3 approacches: 1 the dependency of the transfection method by using 9 transfection reagents, 2 the influence of the time delay of the activity of transfected gene on the correlation by using HeLa Tet-On, 3 the influence of the electroporation as the physical transfection method on the correlation. All results suggested that the nuclear transfer across the nuclear membrane is the rate limiting step o gene expression. By using this method, we investigate the activity of promoters by comparing the time period of cell division and onset of gene expression. As the result, the time period had statistically significance depend on the promoter. Our method is able to measure the cellular dynamics by quantify the activity of promoter.

在这项研究中，我们通过测量EGFP(或Venus)感染的HeLa细胞的时间进程来阐明细胞分裂和基因表达开始之间的强烈相关性。接下来，我们从3个方面探讨了这种现象的共性：1使用9种转染剂的转染法的依赖性，2用HeLa Tet-On法检测基因活性的延时对相关性的影响，3电穿孔作为物理转染法对相关性的影响。这些结果表明，跨核膜的核转移是基因表达的速率限制步骤。利用这种方法，我们通过比较细胞分裂和基因表达开始的时间段来研究启动子的活性。结果表明，不同启动子的时间段有统计学意义。我们的方法能够通过量化启动子的活性来测量细胞动力学。

项目成果

細胞運動評価チップを用いた細胞運動に関わるキノームの解析

利用细胞运动评估芯片分析与细胞运动相关的激酶组

• 发表时间: 2009
• 作者: 島内寿徳;Huong Thi Vu;馬越大;久保井亮一;藤田聡史;Akiko Suzuki(contribution as the first author);Akiko Suzuki(contribution as the first author);長崎玲子
• 通讯作者: 長崎玲子

Onset timing of transient gene expression depends on cell division

瞬时基因表达的开始时间取决于细胞分裂

• 发表时间: 2010
• 期刊: Journal of Bioscience and Bioengineering Vo.109 Issue:1
• 作者: HAKAMADA;K;FUJITA;S;MIYAKE;J
• 通讯作者: J

遺伝子発現と細胞分裂の相関性の一般性に関する検討

检查基因表达与细胞分裂之间相关性的普遍性

• 发表时间: 2010
• 作者: Kazumi Hakamada;Jun Miyake
• 通讯作者: Jun Miyake

Cell division is the key of the onset timing of the transfected gene expression

细胞分裂是转染基因表达起始时间的关键

• 发表时间: 2009
• 作者: Kazumi Hakamada;Jun Miyake
• 通讯作者: Jun Miyake

A system for analyzing the event timing profile of single cells by using a model of neurite maturation in PC12D cells

使用 PC12D 细胞中神经突成熟模型分析单细胞事件时间分布的系统

• 发表时间: 2009
• 期刊: Biosensors & Bioelectronics Vol.24 Issue:5
• 作者: YAMADA;S;HAKAMADA;K;MUNEKATA;T;et al.
• 通讯作者: et al.