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Targeted Detection of G‐Quadruplexes in Cellular RNAs
Author(s) -
Kwok Chun Kit,
Balasubramanian Shankar
Publication year - 2015
Publication title -
angewandte chemie international edition
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 5.831
H-Index - 550
eISSN - 1521-3773
pISSN - 1433-7851
DOI - 10.1002/anie.201500891
Subject(s) - rna , reverse transcriptase , nucleic acid , transcription (linguistics) , computational biology , biology , microbiology and biotechnology , non coding rna , genetics , gene , linguistics , philosophy
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.