Molecular and Cellular Oncogenesis Program, The Wistar Institute, Philadelphia, Pennsylvania.
J Cell Physiol. 2014 Mar;229(3):259-65. doi: 10.1002/jcp.24445.
For a gene to be expressed, the functions of multiple molecular machines must be coordinated at the site of transcription. To understand the role of nuclear organization in transcription, it is necessary to visualize the dynamic interactions of regulatory factors with chromatin and RNA. It is currently possible to localize individual transcription sites in single living mammalian cells by engineering reporter gene constructs to include sequence elements which permit the visualization of nucleic acids in vivo. Upon stable integration, these transgenes form chromatinized arrays, which can be imaged during activation to obtain high-resolution quantitative information about transcriptional dynamics. Modeling can suggest new hypotheses about gene regulation, which can be tested both in the single-cell imaging system and at endogenous genes. This gene-specific imaging strategy has the potential to reveal regulatory mechanisms, which would be difficult to imagine outside of single living cells.
为了实现基因表达,多个分子机器的功能必须在转录部位协调。为了了解核组织在转录中的作用,有必要可视化调控因子与染色质和 RNA 的动态相互作用。目前,可以通过工程报告基因构建来定位单个转录位点,这些构建包括允许在体内可视化核酸的序列元件。在稳定整合后,这些转基因形成染色质化的阵列,可以在激活过程中进行成像,以获得关于转录动力学的高分辨率定量信息。建模可以提出关于基因调控的新假设,这些假设可以在单细胞成像系统和内源性基因中进行测试。这种基因特异性成像策略有可能揭示调控机制,这些机制在单细胞之外是很难想象的。