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运用荧光信号放大和背景降低技术对DNA和RNA进行活细胞成像。

Live cell imaging of DNA and RNA with fluorescent signal amplification and background reduction techniques.

作者信息

Lu Song, Hou Yu, Zhang Xian-En, Gao Yunhua

机构信息

Center for Advanced Measurement Science, National Institute of Metrology, Beijing, China.

Institute of Synthetic Biology, Shenzhen Institute of Advanced Technology, Chinese Academy of Sciences, Shenzhen, China.

出版信息

Front Cell Dev Biol. 2023 Jun 5;11:1216232. doi: 10.3389/fcell.2023.1216232. eCollection 2023.

Abstract

Illuminating DNA and RNA dynamics in live cell can elucidate their life cycle and related biochemical activities. Various protocols have been developed for labeling the regions of interest in DNA and RNA molecules with different types of fluorescent probes. For example, CRISPR-based techniques have been extensively used for imaging genomic loci. However, some DNA and RNA molecules can still be difficult to tag and observe dynamically, such as genomic loci in non-repetitive regions. In this review, we will discuss the toolbox of techniques and methodologies that have been developed for imaging DNA and RNA. We will also introduce optimized systems that provide enhanced signal intensity or low background fluorescence for those difficult-to-tag molecules. These strategies can provide new insights for researchers when designing and using techniques to visualize DNA or RNA molecules.

摘要

阐明活细胞中的DNA和RNA动态变化可以揭示它们的生命周期及相关生化活动。人们已经开发出各种方案,用不同类型的荧光探针标记DNA和RNA分子中的感兴趣区域。例如,基于CRISPR的技术已被广泛用于基因组位点成像。然而,一些DNA和RNA分子仍然难以标记和动态观察,比如非重复区域的基因组位点。在这篇综述中,我们将讨论已开发出的用于DNA和RNA成像的技术和方法工具箱。我们还将介绍优化系统,这些系统为那些难以标记的分子提供增强的信号强度或低背景荧光。这些策略可为研究人员在设计和使用可视化DNA或RNA分子的技术时提供新的见解。

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