Centre for Gene Regulation and Expression, School of Life Sciences, University of Dundee, Dundee DD1 5EH, UK.
FingerPrints Proteomics Facility, School of Life Sciences, University of Dundee, Dundee DD1 5EH, UK.
Open Biol. 2020 Feb;10(2):190306. doi: 10.1098/rsob.190306. Epub 2020 Feb 26.
Eukaryotic messenger RNA (mRNA) is modified by the addition of an inverted guanosine cap to the 5' triphosphate. The cap guanosine and initial transcribed nucleotides are further methylated by a series of cap methyltransferases to generate the mature cap structures which protect RNA from degradation and recruit proteins involved in RNA processing and translation. Research demonstrating that the cap methyltransferases are regulated has generated interest in determining the methylation status of the mRNA cap structures present in cells. Here, we present CAP-MAP: cap analysis protocol with minimal analyte processing, a rapid and sensitive method for detecting cap structures present in mRNA isolated from tissues or cultured cells.
真核信使 RNA（mRNA）通过在 5'三磷酸上加一个反向鸟苷帽来修饰。帽鸟苷和初始转录核苷酸进一步被一系列帽甲基转移酶甲基化，生成成熟的帽结构，保护 RNA 免受降解，并募集参与 RNA 加工和翻译的蛋白质。研究表明，帽甲基转移酶受到调控，这引起了人们对确定细胞中存在的 mRNA 帽结构的甲基化状态的兴趣。在这里，我们介绍 CAP-MAP：带有最小分析物处理的帽分析方案，这是一种快速、灵敏的方法，用于检测从组织或培养细胞中分离的 mRNA 中存在的帽结构。
