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Related Experiment Video

Updated: Jun 22, 2025

Single-Molecule Fluorescence Visualization of DNA Polymerase Dynamics at G-Quadruplexes
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Targeting specific DNA G-quadruplexes with CRISPR-guided G-quadruplex-binding proteins and ligands.

Geng Qin1,2, Zhenqi Liu1,2, Jie Yang1,2

  • 1Laboratory of Chemical Biology and State Key Laboratory of Rare Earth Resource Utilization, Changchun Institute of Applied Chemistry, Chinese Academy of Sciences, Changchun, P. R. China.

Nature Cell Biology
|July 3, 2024
PubMed
Summary
This summary is machine-generated.

Researchers developed a CRISPR-based method to precisely control DNA G-quadruplexes (G4s). This technique enables targeted G4 folding for studying their roles in health and disease and for developing new therapies.

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Area of Science:

  • Molecular Biology
  • Genetics
  • Biotechnology

Background:

  • DNA G-quadruplexes (G4s) are crucial in biological processes and diseases.
  • Existing technologies lack precision in manipulating specific G4 structures for functional studies and therapeutic development.

Purpose of the Study:

  • To develop a novel technology for targeted manipulation of specific DNA G-quadruplex folding.
  • To investigate the biological functions of specific G4s using precise manipulation techniques.
  • To explore potential therapeutic applications by controlling G4 structures.

Main Methods:

  • Utilized CRISPR technology combined with G4-stabilizing proteins (e.g., nucleolin) and ligands.
  • Engineered catalytically inactive Cas9 fused with nucleolin to target and stabilize specific G4s in genomic loci.
  • Employed CRISPR to enhance the selectivity of G4-binding compounds (pyridodicarboxamide, pyridostatin) for intra-G4 targeting.

Main Results:

  • Successfully stabilized G4s in the MYC oncogene promoter, Itga7 gene promoter, and telomeres using CRISPR-nucleolin fusion.
  • Demonstrated that targeted G4 stabilization leads to significant cellular effects: proliferation arrest, inhibited myoblast differentiation, and cell senescence.
  • Achieved enhanced selectivity of G4 ligands, enabling more precise investigation of de novo G4 functions compared to traditional methods.

Conclusions:

  • The developed CRISPR-based system provides unprecedented precision in manipulating DNA G-quadruplex folding at specific genomic sites.
  • This technology offers a powerful tool for dissecting the functional roles of G4s in various biological contexts.
  • The findings pave the way for advancing G4-related research and developing targeted therapeutic strategies for diseases involving G4s.