U2.3 前体小核RNA加工分析
U2.3 Precursor Small Nuclear RNA Processing Assay.
作者信息
Lin Chan, Feng Yujie, Peng Xueyan, Wu Jiaming, Wang Weili, Liu Yunfeng
机构信息
State Key Laboratory for Conservation and Utilization of Subtropical Agro-bioresources, College of Life Science and Technology, Guangxi University, 100 Daxue Road, Nanning, Guangxi 530004, China.
出版信息
Bio Protoc. 2021 Sep 5;11(17):e4142. doi: 10.21769/BioProtoc.4142.
Small nuclear RNAs (snRNAs) are vital for eukaryotic cell activities and play important roles in pre-mRNA splicing. The molecular mechanism underlying the transcription of snRNA, regulated via upstream/downstream cis-elements and relevant trans-elements, has been investigated in detail using cell-free extracts. However, the processing of precursor snRNA (pre-snRNA), which is required by 3' end maturation of pre-snRNA, remains unclear as a proper processing assay is difficult to develop . Here, we present an method using synthetic labeled RNA as substrates to study the 3' cleavage of pre-snRNA.
小核RNA（snRNA）对真核细胞活动至关重要，并且在信使前体核糖核酸（pre-mRNA）剪接中发挥重要作用。通过无细胞提取物已详细研究了snRNA转录的分子机制，该机制通过上游/下游顺式元件和相关反式元件进行调控。然而，由于难以开发合适的加工检测方法，前体snRNA（pre-snRNA）的加工（这是pre-snRNA 3'末端成熟所必需的）仍不清楚。在这里，我们提出了一种使用合成标记RNA作为底物来研究pre-snRNA 3'切割的方法。
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