Fok Ezio T, Scholefield Janine, Fanucchi Stephanie, Mhlanga Musa M
Gene Expression & Biophysics Group, Biosciences, CSIR, Pretoria, Gauteng, South Africa.
Division of Chemical, Systems & Synthetic Biology, Institute for Infectious Disease & Molecular Medicine, Faculty of Health Sciences, University of Cape Town, Cape Town, Western Cape, South Africa.
Epigenomics. 2017 Oct;9(10):1317-1327. doi: 10.2217/epi-2017-0062. Epub 2017 Sep 6.
Long noncoding RNAs (lncRNAs) have been implicated in many biological processes. However, due to the unique nature of lncRNAs and the consequential difficulties associated with their characterization, there is a growing disparity between the rate at which lncRNAs are being discovered and the assignment of biological function to these transcripts. Here we present a molecular biology toolbox equipped to help dissect aspects of lncRNA biology and reveal functionality. We outline an approach that begins with a broad survey of genome-wide, high-throughput datasets to identify potential lncRNA candidates and then narrow the focus on specific methods that are well suited to interrogate the transcripts of interest more closely. This involves the use of imaging-based strategies to validate these candidates and observe the behaviors of these transcripts at single molecule resolution in individual cells. We also describe the use of gene editing tools and interactome capture techniques to interrogate functionality and infer mechanism, respectively. With the emergence of lncRNAs as important molecules in healthy and diseased cellular function, it remains crucial to deepen our understanding of their biology.
长链非编码RNA（lncRNAs）已被证明参与许多生物学过程。然而，由于lncRNAs的独特性质以及随之而来的对其进行表征的困难，lncRNAs的发现速度与赋予这些转录本生物学功能之间的差距越来越大。在此，我们展示了一个分子生物学工具箱，旨在帮助剖析lncRNA生物学的各个方面并揭示其功能。我们概述了一种方法，该方法首先对全基因组高通量数据集进行广泛调查，以识别潜在的lncRNA候选物，然后将重点缩小到更适合更深入研究感兴趣转录本的特定方法上。这涉及使用基于成像的策略来验证这些候选物，并在单个细胞中以单分子分辨率观察这些转录本的行为。我们还描述了分别使用基因编辑工具和相互作用组捕获技术来研究功能和推断机制。随着lncRNAs成为健康和患病细胞功能中的重要分子，加深我们对其生物学的理解仍然至关重要。
