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.
Highlights
The G-quadruplex (G4) is a non-canonical nucleic acid structure which regulates important cellular processes
Described 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
Strong evidence is provided for G4 formation in full-length cellular human telomerase RNA, offering important insights into its cellular function
Summary
The G-quadruplex (G4) is a non-canonical nucleic acid structure which regulates important cellular processes. Cellular transcripts are of lower abundance, it is challenging to adapt existing approaches to probe the formation and location of G4s in full-length cellular RNAs. we show that RNA G4-mediated reverse transcriptase stalling (RTS) can be rationally controlled in a cation- and G4 ligand-dependent fashion (Figure 1).
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