Kwok Chun Kit, Balasubramanian Shankar
The University of Cambridge, Department of Chemistry, Lensfield Road, Cambridge, CB2 1EW (UK).
Angew Chem Int Ed Engl. 2015 Jun 1;54(23):6751-4. doi: 10.1002/anie.201500891. Epub 2015 Apr 23.
The G-quadruplex (G4) is a non-canonical nucleic acid structure which regulates important cellular processes. RNA G4s have recently been shown to exist in human cells and be biologically significant. Described herein is a new approach to detect and map RNA G4s in cellular transcripts. This method exploits the specific control of RNA G4-cation and RNA G4-ligand interactions during reverse transcription, by using a selective reverse transcriptase to monitor RNA G4-mediated reverse transcriptase stalling (RTS) events. Importantly, a ligation-amplification strategy is coupled with RTS, and enables detection and mapping of G4s in important, low-abundance cellular RNAs. Strong evidence is provided for G4 formation in full-length cellular human telomerase RNA, offering important insights into its cellular function.
G-四链体（G4）是一种非经典核酸结构，可调节重要的细胞过程。最近研究表明，RNA G4存在于人类细胞中且具有生物学意义。本文描述了一种检测和定位细胞转录本中RNA G4的新方法。该方法利用逆转录过程中RNA G4-阳离子和RNA G4-配体相互作用的特异性控制，通过使用选择性逆转录酶监测RNA G4介导的逆转录酶停滞（RTS）事件。重要的是，连接扩增策略与RTS相结合，能够检测和定位重要的低丰度细胞RNA中的G4。为全长细胞人端粒酶RNA中G4的形成提供了有力证据，为其细胞功能提供了重要见解。
