title: Targeted Detection of G-Quadruplexes in Cellular RNAs.
abstract: 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.
authors.affiliation: The University of Cambridge, Department of Chemistry, Lensfield Road, Cambridge, CB2 1EW (UK).
authors.forename: Chun Kit
authors.initials: CK
authors.lastname: Kwok
authors.affiliation: The University of Cambridge, Department of Chemistry, Lensfield Road, Cambridge, CB2 1EW (UK). sb10031@cam.ac.uk.
authors.forename: Shankar
authors.initials: S
authors.lastname: Balasubramanian
mesh_terms: D054856:G-Quadruplexes; D006367:HeLa Cells; D006801:Humans; D015394:Molecular Structure; D012313:RNA / Q000737:chemistry* / Q000378:metabolism; D019098:Telomerase / Q000737:chemistry* / Q000378:metabolism
chemical_list: C422952:telomerase RNA; D012313:RNA; D019098:Telomerase
keywords: G-quadruplexes; RNA structures; cations; ligand effects; reverse transcription; telomerase RNA