Department of Biology of the Cell Nucleus, Institute of Molecular Genetics of the Czech Academy of Sciences, Vídeňská 1083, 142 20 Prague, Czech Republic.
Faculty of Science, Charles University, Albertov 6, 128 00 Prague, Czech Republic.
Int J Mol Sci. 2021 Jun 22;22(13):6694. doi: 10.3390/ijms22136694.
Classical models of gene expression were built using genetics and biochemistry. Although these approaches are powerful, they have very limited consideration of the spatial and temporal organization of gene expression. Although the spatial organization and dynamics of RNA polymerase II (RNAPII) transcription machinery have fundamental functional consequences for gene expression, its detailed studies have been abrogated by the limits of classical light microscopy for a long time. The advent of super-resolution microscopy (SRM) techniques allowed for the visualization of the RNAPII transcription machinery with nanometer resolution and millisecond precision. In this review, we summarize the recent methodological advances in SRM, focus on its application for studies of the nanoscale organization in space and time of RNAPII transcription, and discuss its consequences for the mechanistic understanding of gene expression.
经典的基因表达模型是基于遗传学和生物化学构建的。尽管这些方法非常强大，但它们对基因表达的空间和时间组织考虑得非常有限。尽管 RNA 聚合酶 II（RNAPII）转录机制的空间组织和动态对基因表达具有根本的功能意义，但由于经典的明场显微镜的限制，其详细研究长期以来一直受到阻碍。超分辨率显微镜（SRM）技术的出现使得能够以纳米分辨率和毫秒精度可视化 RNAPII 转录机制。在这篇综述中，我们总结了 SRM 的最新方法学进展，重点介绍了其在研究 RNAPII 转录的纳米级空间和时间组织中的应用，并讨论了其对基因表达的机制理解的影响。
