Highly Resolved Detection of Long Non-coding RNAs In Situ.
Highly Resolved Detection of Long Non-coding RNAs In Situ.
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DOI：
10.1007/978-1-0716-1697-0_12
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发表时间：
2021
期刊：
影响因子：
--
通讯作者：
Kalantry S
中科院分区：
文献类型：
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作者：
Trotter M;Harris C;Cloutier M;Samanta M;Kalantry S
长链非编码RNA（lncRNA）被认为在许多基于DNA的过程中发挥作用，最值得注意的是转录。原位检测lncRNA可以深入了解其功能。荧光原位杂交（FISH）能够检测单个细胞内的特定核酸序列，包括lncRNA。目前的RNA FISH技术可以告知RNA转录本的定位和表达水平。连同显微镜的进步，这些原位技术现在允许可视化和定量甚至低表达或不稳定的lncRNA。当结合检测相关蛋白质和染色质修饰的免疫荧光，RNA FISH可以提供必要的见解lncRNA的功能。在这里，我们描述了一套完整的协议，以检测，单独或组合，特定的RNA，DNA，蛋白质和组蛋白修饰在单细胞中，在高灵敏度使用传统的荧光显微镜。
Long non-coding RNAs (lncRNAs) have been postulated to function in a number of DNA-based processes, most notably transcription. The detection of lncRNAs in situ can offer insights into their function. Fluorescence in situ hybridization (FISH) enables the detection of specific nucleic acid sequences, including lncRNAs, within individual cells. Current RNA FISH techniques can inform both the localization and expression level of RNA transcripts. Together with advances in microscopy, these in situ techniques now allow for visualization and quantification of even lowly expressed or unstable lncRNAs. When combined with detection of associated proteins and chromatin modifications by immunofluorescence, RNA FISH can lend essential insights into lncRNA function. Here, we describe an integrated set of protocols to detect, individually or in combination, specific RNAs, DNAs, proteins, and histone modifications in single cells at high sensitivity using conventional fluorescence microscopy.