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Probing pluripotency gene regulatory networks with quantitative live cell imaging
Computational and Structural Biotechnology Journal ( IF 4.4 ) Pub Date : 2020-09-20 , DOI: 10.1016/j.csbj.2020.09.025
Anne L. Plant , Michael Halter , Jeffrey Stinson

Live cell imaging uniquely enables the measurement of dynamic events in single cells, but it has not been used often in the study of gene regulatory networks. Network components can be examined in relation to one another by quantitative live cell imaging of fluorescent protein reporter cell lines that simultaneously report on more than one network component. A series of dual-reporter cell lines would allow different combinations of network components to be examined in individual cells. Dynamical information about interacting network components in individual cells is critical to predictive modeling of gene regulatory networks, and such information is not accessible through omics and other end point techniques. Achieving this requires that gene-edited cell lines are appropriately designed and adequately characterized to assure the validity of the biological conclusions derived from the expression of the reporters. In this brief review we discuss what is known about the importance of dynamics to network modeling and review some recent advances in optical microscopy methods and image analysis approaches that are making the use of quantitative live cell imaging for network analysis possible. We also discuss how strategies for genetic engineering of reporter cell lines can influence the biological relevance of the data.



中文翻译:

用定量活细胞成像探索多能性基因调控网络

活细胞成像独特地能够测量单个细胞中的动态事件,但在基因调节网络的研究中并未经常使用它。可以通过对同时报告多个网络组件的荧光蛋白报告基因细胞系进行定量活细胞成像,从而相互检查网络组件。一系列的双重报告者细胞系将允许在单个细胞中检查网络组件的不同组合。有关单个细胞中相互作用的网络组件的动态信息对于基因调节网络的预测建模至关重要,并且此类信息无法通过组学和其他端点技术获得。为此,需要对基因编辑的细胞系进行适当的设计和适当的表征,以确保从报道基因的表达得出的生物学结论的有效性。在这篇简短的综述中,我们讨论了动力学对网络建模的重要性,并回顾了光学显微镜方法和图像分析方法的一些最新进展,这些进展使得使用定量活细胞成像进行网络分析成为可能。我们还将讨论报告细胞系遗传工程的策略如何影响数据的生物学相关性。在这篇简短的综述中,我们讨论了动力学对网络建模的重要性,并回顾了光学显微镜方法和图像分析方法的一些最新进展,这些进展使得使用定量活细胞成像进行网络分析成为可能。我们还将讨论报告细胞系遗传工程的策略如何影响数据的生物学相关性。在这篇简短的综述中,我们讨论了动力学对网络建模的重要性的已知知识,并回顾了光学显微镜方法和图像分析方法的一些最新进展,这些进展使得利用定量活细胞成像进行网络分析成为可能。我们还将讨论报告细胞系遗传工程的策略如何影响数据的生物学相关性。

更新日期:2020-09-20
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