Mammalian genomes encode thousands of long noncoding RNAs (lncRNAs) that are often expressed in a tissue and cell specific manner. Therefore, a reporter that can faithfully reflect the expression or activity of lncRNAs can provide tools useful not only for uncovering the regulators of lncRNAs, but also for tracking cell fate and disease status. Here, we design a sgRNA precursor in an intron (GRIT) strategy that can monitor the promoter activity of lncRNAs. We used this strategy to report the expression of Lncenc1 and Neat1 in mouse embryonic stem cells (ESCs). Furthermore, we show that GRIT may be used to track differentiation status of stem cells. We anticipate that GRIT will be applicable in dissecting regulatory mechanisms underlying the transcription of lncRNAs, tracking cell fate switch during differentiation or disease progression and integrating the promoter activity of various RNAs for synthetic biology applications.

中文翻译：

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通过内含子中tRNA旁接的sgRNA敲入来监测长非编码RNA的启动子活性和干细胞分化

哺乳动物基因组编码数千个通常以组织和细胞特异性方式表达的长非编码RNA（lncRNA）。因此，能够忠实地反映lncRNA的表达或活性的报道者可以提供工具，这些工具不仅可用于发现lncRNA的调节子，而且可用于追踪细胞命运和疾病状态。在这里，我们设计了一种内含子（GRIT）策略中的sgRNA前体，该策略可以监测lncRNA的启动子活性。我们使用这种策略来报告Lncenc1和Neat1在小鼠胚胎干细胞（ESC）中的表达。此外，我们表明，GRIT可用于跟踪干细胞的分化状态。我们预计GRIT将适用于剖析lncRNA转录基础的调控机制，