Yang Yun, Wang Zefeng
CAS Key Lab for Computational Biology, CAS-MPG Partner Institute for Computational Biology, Shanghai, China.
CAS Center for Excellence in Molecular Cell Science, Shanghai Institute for Biological Sciences, Chinese Academy of Sciences, Shanghai, China.
Methods Mol Biol. 2018;1724:107-118. doi: 10.1007/978-1-4939-7562-4_9.
Human transcriptome contains a large number of circular RNAs (circRNAs) that are mainly produced by back splicing of pre-mRNA. Here we describe a minigene reporter system containing a single exon encoding split GFP in reverse order, which can be efficiently back spliced to produce a circRNA encoding intact GFP gene. This simple reporter system can be adopted to study how different cis-elements and trans-factors affect circRNA production, and also can serve as a reliable system to measure the activity of IRES-mediated translation. Therefore this system can serve as a platform for mechanistic studies on the circRNA biogenesis and its function.
人类转录组包含大量环状RNA（circRNA），它们主要由前体mRNA的反向剪接产生。在这里，我们描述了一种小基因报告系统，该系统包含一个以反向顺序编码分裂绿色荧光蛋白（GFP）的单个外显子，其可以有效地进行反向剪接以产生编码完整GFP基因的circRNA。这个简单的报告系统可用于研究不同的顺式元件和反式因子如何影响circRNA的产生，也可作为测量内部核糖体进入位点（IRES）介导的翻译活性的可靠系统。因此，该系统可作为circRNA生物发生及其功能机制研究的平台。
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