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Related Experiment Video

Updated: Dec 17, 2025

Author Spotlight: Characterizing DNA G-Quadruplex by Bis-3-Chloropiperidine Based Chemical Mapping
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Detection of cellular G-quadruplex by using a loop structure as a structural determinant.

Hisao Masai1, Yutaka Kanoh1, Naoko Kakusho1

  • 1Department of Basic Medical Sciences, Tokyo Metropolitan Institute of Medical Science, Kamikitazawa, Setagaya-ku, Tokyo, 156-8506, Japan.

Biochemical and Biophysical Research Communications
|June 27, 2020
PubMed
Summary

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This study demonstrates a new method to detect G-quadruplex (G4) structures within cells. By inserting a unique DNA sequence, researchers can now visualize G4 presence and dynamics in real-time, advancing cellular biology research.

Area of Science:

  • Genomics
  • Molecular Biology
  • Biochemistry

Background:

  • G-quadruplex (G4) structures are increasingly recognized for their roles in biological processes.
  • Directly visualizing G4 structures within living cells has been challenging due to methodological limitations.

Purpose of the Study:

  • To develop and validate a novel, versatile, and non-biased methodology for detecting G-quadruplex structures in vivo.
  • To provide direct evidence for the formation of G4 structures at specific genomic locations in fission yeast.

Main Methods:

  • Utilized Rif1 binding sites (Rif1BS) in fission yeast, known to form G4 structures.
  • Engineered Rif1BS by replacing long single-stranded loops with an I-SceI restriction site.
  • Assessed I-SceI cleavage in vitro and in vivo, correlating it with G4 formation and disruption.
Keywords:
G-quadruplexIn vivo detection of G4LoopRNA-DNA hybridRestriction digestionRif1

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Main Results:

  • In vitro experiments confirmed that I-SceI is not cleaved when G4 structures are formed on duplex Rif1BS DNA.
  • Cleavage of I-SceI was observed when G4 structures were absent, indicating successful G4 detection.
  • Demonstrated in vivo cleavage of I-SceI in fission yeast cells, providing evidence for cellular G4 presence.

Conclusions:

  • The developed method offers a reliable platform for detecting G4 structures in vivo.
  • This strategy enables genome-wide analysis of cellular G4 formation and disruption dynamics.
  • The findings pave the way for a deeper understanding of G4 roles in cellular functions.