在活真核细胞中对 DNA 和 RNA 的成像以揭示基因表达的时空动态。
Imaging of DNA and RNA in Living Eukaryotic Cells to Reveal Spatiotemporal Dynamics of Gene Expression.
机构信息
Department of Anatomy and Structural Biology, Albert Einstein College of Medicine, Bronx, New York 10461, USA; email:
Janelia Research Campus, Howard Hughes Medical Institute, Ashburn, Virginia 20147, USA.
出版信息
Annu Rev Biochem. 2020 Jun 20;89:159-187. doi: 10.1146/annurev-biochem-011520-104955. Epub 2020 Mar 16.
Abstract
This review focuses on imaging DNA and single RNA molecules in living cells to define eukaryotic functional organization and dynamic processes. The latest advances in technologies to visualize individual DNA loci and RNAs in real time are discussed. Single-molecule fluorescence microscopy provides the spatial and temporal resolution to reveal mechanisms regulating fundamental cell functions. Novel insights into the regulation of nuclear architecture, transcription, posttranscriptional RNA processing, and RNA localization provided by multicolor fluorescence microscopy are reviewed. A perspective on the future use of live imaging technologies and overcoming their current limitations is provided.
摘要
本文综述了在活细胞中成像 DNA 和单 RNA 分子以定义真核生物功能组织和动态过程的研究进展。本文讨论了实时可视化单个 DNA 基因座和 RNA 的最新技术进展。单分子荧光显微镜提供了时空分辨率，揭示了调节基本细胞功能的机制。多色荧光显微镜为核架构、转录、转录后 RNA 加工和 RNA 定位的调控提供了新的见解。本文还展望了活细胞成像技术的未来应用，并探讨了如何克服其当前的局限性。
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本文引用的文献
1
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Mol Cell. 2019 Oct 17;76(2):295-305. doi: 10.1016/j.molcel.2019.09.016. Epub 2019 Oct 8.
2
Evaluating phase separation in live cells: diagnosis, caveats, and functional consequences.评估活细胞中的相分离：诊断、注意事项和功能后果。
Genes Dev. 2019 Dec 1;33(23-24):1619-1634. doi: 10.1101/gad.331520.119. Epub 2019 Oct 8.
3
Fast objective coupled planar illumination microscopy.快速客观耦合平面照明显微镜。
Nat Commun. 2019 Oct 2;10(1):4483. doi: 10.1038/s41467-019-12340-0.
4
Rational Design of Fluorogenic and Spontaneously Blinking Labels for Super-Resolution Imaging.用于超分辨率成像的荧光生成和自发闪烁标记的合理设计
ACS Cent Sci. 2019 Sep 25;5(9):1602-1613. doi: 10.1021/acscentsci.9b00676. Epub 2019 Sep 5.
5
Visualizing RNA dynamics in live cells with bright and stable fluorescent RNAs.用明亮稳定的荧光 RNA 可视化活细胞中的 RNA 动态。
Nat Biotechnol. 2019 Nov;37(11):1287-1293. doi: 10.1038/s41587-019-0249-1. Epub 2019 Sep 23.
6
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Cell. 2019 Sep 19;179(1):147-164.e20. doi: 10.1016/j.cell.2019.08.050.
7
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9
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Science. 2019 Sep 20;365(6459):1301-1305. doi: 10.1126/science.aax7852. Epub 2019 Sep 5.
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