通过逆转录聚合酶链反应（RT-PCR）确认转录通读事件
Confirmation of Transcriptional Read-Through Events by RT-PCR.
作者信息
Shi Xinrui, Qin Fujun, Li Hui
机构信息
Department of Biochemistry and Molecular Genetics, School of Medicine, University of Virginia, Charlottesville, VA, USA.
Department of Pathology, School of Medicine, University of Virginia, Charlottesville, VA, USA.
出版信息
Methods Mol Biol. 2020;2079:177-186. doi: 10.1007/978-1-4939-9904-0_14.
Chimeric RNAs can be formed by trans-splicing from different transcripts or cis-splicing of adjacent genes (cis-SAGe). Cis-SAGe results from read-through transcription of two neighbor genes. To investigate the mechanisms underlying intergenic splicing of adjacent genes, it is important to develop an assay to detect transcriptional read-through. Here, we describe a general RT-PCR based method to confirm the process for cis-SAGe candidates. In this method, we use PCR to amplify cDNA that is reverse transcribed from the read-through precursor mRNA. The result provides a foundation for further downstream mechanistic studies.
嵌合RNA可通过不同转录本的反式剪接或相邻基因的顺式剪接（顺式SAGe）形成。顺式SAGe是由两个相邻基因的通读转录产生的。为了研究相邻基因间剪接的潜在机制，开发一种检测转录通读的方法很重要。在此，我们描述了一种基于RT-PCR的通用方法来确认顺式SAGe候选物的过程。在这种方法中，我们使用PCR扩增从通读前体mRNA逆转录而来的cDNA。该结果为进一步的下游机制研究奠定了基础。
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