相关实验视频

Updated: Jul 16, 2025

Applying an Inducible Expression System to Study Interference of Bacterial Virulence Factors with Intracellular Signaling

Applying an Inducible Expression System to Study Interference of Bacterial Virulence Factors with Intracellular Signaling

Published on: June 25, 2015

在Tet-On系统中,统一的转基因激活取决于持续的rtTA表达

Jun Otomo¹, Knut Woltjen¹, Hidetoshi Sakurai¹

¹Center for iPS Cell Research and Application (CiRA), Kyoto University, Sakyo-ku, Kyoto 606-8507, Japan.

|September 13, 2023

概括

补充反向四环素交换激活剂 (rtTA) 确保在人类诱导多能干细胞 (hiPSCs) 中均的Tet-On基因表达. 将rtTA与胺素选择联系起来,可以提高均性,通过Tet-MyoD实现高效的肌原分化.

科学领域:

干细胞生物学干细胞生物学
分子生物学分子生物学
基因规则基因规则

背景情况:

环素诱导 (Tet-On) 系统对于人类诱导多能干细胞 (hiPSCs) 中调节基因表达至关重要.
不统一的转基因激活仍然是高PSC中的Tet-On系统应用的重大挑战.

研究的目的:

通过使用Tet-On系统,增强hiPSC中均的转基因激活.
研究抗生素选择标记物对rtTA表达和Tet-On系统性能的影响.
开发一个优化的Tet-On系统,以实现高PSCs的高效肌性分化.

主要方法:

在多克隆的hiPSC种群中补充反向四环素交互激活剂 (rtTA).
使用与内部核糖体进入部位 (IRES) 相关的抗生素选择标记 (胺与新胺) 进行rtTA表达.
设计一个四环素诱导的MyoD载体 (Tet-MyoD) 用于肌体差异化研究.

主要成果:

补充rtTA在hiPSC中实现了统一的Tet-On转基因激活.
当通过IRES连接胺素选择标记时,与neomycin相比,它产生了更统一的rtTA表达.
优化的Tet-MyoD系统证明了hiPSCs的高效,强大和可重复的定向肌原分化.

结论:

优化的rtTA补充和选择标记的选择可以克服hiPSCs内的Tet-On系统中的不均性.
素选择为均的rtTA表达提供了优势,简化了hiPSC转基因.
开发的Tet-MyoD系统提供了一个强大的工具,用于控制hiPSCs的肌原分化.

关键词:

细胞生物学细胞生物学生物科学中的方法论.干细胞研究的研究.

更多相关视频

Use of the TetON System to Study Molecular Mechanisms of Zebrafish Regeneration

Use of the TetON System to Study Molecular Mechanisms of Zebrafish Regeneration

Published on: June 25, 2015

Live Imaging of Innate Immune and Preneoplastic Cell Interactions Using an Inducible Gal4/UAS Expression System in Larval Zebrafish Skin

Live Imaging of Innate Immune and Preneoplastic Cell Interactions Using an Inducible Gal4/UAS Expression System in Larval Zebrafish Skin

Published on: February 3, 2015

相关概念视频

Reporter Genes

Reporter Genes

Reporter genes are a type of protein-coding gene that are often tagged to a gene of interest. Once inside a target cell, reporter genes usually produce visually identifiable characteristics like fluorescence and luminescence when expressed along with the gene of interest. Thus, reporter genes “report” the presence or absence of genes of interest in an organism, determine the gene expression pattern, or track the physical location of a DNA segment or protein in the cell.

Transgenic Organisms

Transgenic Organisms

Transgenic Plants

Transgenic Plants

Recombinant DNA technology called transgenesis is often used to add a foreign gene or remove a detrimental gene from an organism. Such genetically modified organisms are called transgenic organisms.
The first-ever transgenic plant was a tobacco plant developed in 1983 that showed resistance against the tobacco mosaic virus. Since then, many transgenic plants have been developed and commercialized for improving the agricultural, ornamental, and horticultural value of a crop plant. Transgenic...